This study examined the differentiation character and pluripotency of mesenchymal stem cells (MSCs) under different conditions. Adult MSCs were initially isolated from the bone marrow of rats, cultured in vitro and identified by flow cytometry. After MSCs were transferred to osteogenic and adipogenic medium respectively, the morphological characterization of induced cells was observed. The expression of marker genes was detected by RT-PCR analysis. Then MSCs were transfected with lentiviral vectors pGC-FU-Sox9-EGFP. Enhanced green fluorescence protein (EGFP) expression and transfection efficiency were determined by fluorescence microscopy. The results demonstrated that EGFP caused no effect on the multilineage potential of adult MSCs. Sox9 gene expression of high level was maintained stable in the transfected MSCs and induced MSCs to differentiate into chondrocytes. Aggracan was positive in chondrogenic lineages and the expression of aggracan and type collagen II was significantly increased during MSCs chondrogenic differentiation. It was concluded that Sox9 gene-modified adult MSCs may be promising candidate cells for further studies on tissue engineering. EGFP facilitates the research on MSCs physiological behavior and application in tissue engineering during differentiation both in vitro and in vivo.

Tetrahydrobiopterin (BH4) is an essential cofactor for all three nitric oxide synthase (NOS isoforms), which plays an important role in vascular diseases. GTP cyclohydrolase 1 (GCH 1) is the first-step and rate-limiting enzyme for BH4 biosynthesis in its de novo pathway. Common GCH1 gene variant C+243T in the 3′-untranslated region predicts NO excretion. The present study examined the predictive role of GCH 1 gene 3′-UTR C+243T variant in the long-term outcome of ischemic stroke. A total of 142 patients with first-onset ischemic stroke were recruited and detected for genotype of GCH1 3′-UTR C+243T by a TaqMan SNP Genotyping assay. Subsequent vascular events and death were determined over a 5-year follow-up period. The frequency of GCH1 3′-UTR +243 C/T or T/T genotype was significantly increased in patients with endpoint events as compared with those without events (74% vs 57.8%, P=0.06). Cox regression survival analysis indicated that an increased probability of death or new vascular events was found in patients with GCH1 3′-UTR +243 C/T or T/T genotype compared with those with GCH1 3′-UTR C/C genotype (40.6% vs 25.5%), GCH1 3′-UTR +243 C/T or T/T genotype relative to GCH1 3′-UTR C/C genotype was associated with the increased risk of death or vascular events even after adjustment for other risk factors (OR=2.171, 95% CI: 1.066–4.424, P=0.033). It was concluded that GCH1 3′-UTR C+243T variant was an independent predictor of worsening long-term outcomes in patients with first-onset ischemic stroke.

This study examined the predictive value of plasma cystatin C, creatinine and estimated glomerular filtration rate (eGFR) as risk factors for cardiovascular disease in Chinese. Plasma cystatin C and creatinine were measured in 466 coronary heart disease (CHD) patients recruited from 4 hospitals and 349 healthy controls from local communities in Wuhan, China. Cockroft-Gault formula was used to estimate the glomerular filtration rate (GFR) after adjusting for body surface area. With each measure, the study population was divided into quintiles. The results showed that the patients had significantly higher levels of plasma cystatin C, creatinine, and lower level of eGFR than controls. Lower eGFR was associated with a higher risk of cardiovascular events. As compared with the first (highest) quintile, the hazard ratios (and 95% CIs) after multivariate adjustment for CHD were as follows: third quintile, 2.98 (1.54–5.78); fourth quintile, 3.34 (1.58–7.09); fifth quintile, 4.37(1.84–10.35). With higher cystatin C quintiles (≥1.00 mg/L and ≥1.17 mg/L), the hazard ratios for CHD were 2.16 (1.23–3.81) and 2.34 (1.25–4.38), similar to those of creatinine 2.21 (1.21–4.03) and 2.03 (1.07–3.84). However, it was plasma cystatin C not eGFR or creatinine had stronger association with ischemic stroke. The highest quintile had the hazard ratio of 4.51 (1.45–14.08) after multivariate adjustment. It was concluded that plasma cystatin C, associated with renal function, is not an independent risk factor for cardiovascular disease. eGFR is a better risk predictor for CHD than plasma cystatin C and creatinine. But for ischemic stroke, plasma cystatin C is a better risk factor than creatinine and estimated GFR.

This study examined the effect of ischemia-reperfusion injury on the expression of Pim-3 gene in myocardial tissues and their underlying mechanism. Rat models of myocardial ischemia-reperfusion injury were established by ligating the left anterior descending coronary artery of the rats. A total of 30 SD male adult rats were randomly divided into 5 groups: group A (sham operation, n=6); group B (in which the rats were subjected to 15 min of ischemia by ligation of the left anterior descending coronary artery, n=6); group C (in which the rats received 30 min of ischemia, n=6), group D and group E (in which the left anterior descending coronary artery of the rats were ligated for 30 min and then reperfused for 30 min or 120 min, n=6 in each). The left ventricular tissues were removed immediately after the ischemia-reperfusion injury. Neonatal cardiomyocytes were cultured and treated with different concentrations of H₂O₂ (0, 5, 10, 20 μmol/L) or tumor necrosis factor-α (TNF-α, 0, 1, 5, 10 ng/mL). The mRNA and protein expression of Pim-3 gene was determined by using RT-PCR, western blotting and immunohistochemistry. Additionally, neonatal cardiomyocytes were transfected with Pim-3 siRNA, and induced to develop apoptosis by using H₂O₂. The results showed that normal myocardial tissues expressed a quantity of Pim-3 gene mRNA and protein. Ischemia-reperfusion injury could up-regulate the mRNA and protein expression of Pim-3 gene in myocardial tissues. Furthermore, H₂O₂ but not TNF-α up-regulated the Pim-3 gene expression in cultured cardiomyocytes. And Pim-3 silencing failed to strengthen the H₂O₂-inducing apoptosis in cardiomyocytes. It was concluded that ischemia-reperfusion injury up-regulated the Pim-3 gene expression through oxidative stress signaling pathway in myocardial tissues.

This study investigated the effect of cadmium on the telomerase activity, the expression of TERT, c-myc and p53 and the apoptosis of rat hepatocytes. The rats were administrated 5, 10 and 20 μmol/kg cadmium chloride intraperitoneally and sacrificed 48 h after the initial treatment. The telomerase activity of the rat hepatocytes was measured by the telomeric repeat amplification protocol (TRAP), and apoptosis was detected by flow cytometry. The mRNA expressions of TERT, c-myc and p53 were measured by reverse transcription-polymerase chain reaction (RT-PCR). C-myc and P53 proteins were determined by immunochemistry. The results showed that cadmium chloride increased the hepatocellular telomerase activity in a dose-dependant manner and induced the apoptosis of hepatocytes significantly. The value of relative coefficient between the telomerase activity and the apoptosis rate was 0.9398. RT-PCR revealed that specific bands corresponding to the TERT mRNA, c-myc mRNA, and p53 mRNA were displayed at 185, 342 and 538 bp respectively. Cadmium chloride could substantially increase the mRNA expressions of TERT, c-myc and p53 in rat hepatocytes, as compared with control. Moreover, cadmium chloride at the doses of 5, 10 and 20 μmol/kg could increase the content of P53 protein in rat hepatocytes obviously, but only that at the doses of 10 and 20 μmol/kg substantially promoted the c-myc protein level in rat hepatocytes. Our study herein suggested that cadmium may contribute to the carcinogenesis by activating telomerase, and overexpressing the mRNAs of TERT, c-myc and p53, and causing apoptosis of normal cells.

This study examined the association of problem behavior with neurotransmitter deficiency in adolescents, which would provide new insights into behavioral problems. A total of 1259 students of the seventh grade from 4 middle schools in Wuhan city located in the central China were recruited. With the approval of school and parents, they were invited to complete the Youth Self-Report (YSR) questionnaire and Symptom Scale of Neurotransmitter Deficiency (SSND) questionnaire. Pearson’s bivariate correlation analysis showed that the correlation coefficients between each subscale of YSR and SSND ranged from 0.24 to 0.61 with all P<0.01. Canonical correlation analysis indicated that anxiety/depression was interrelated with insufficiency of GABA and 5-HT; aggressive behavior was associated with inadequate GABA; famine of DA influenced the attention problems. It was concluded that neurotransmitter deficiency may cause a series of behavioral and mental problems.

Drug abuse continues to be a serious public health threat worldwide. Most drug abuse prevention research has been conducted with predominantly American or European adolescent populations. Little is known about approaches that work best to prevent the initiation of Chinese adolescent drug use. For targeting risk factors of drug initiation in Chinese adolescents, a school-based health intervention program named “Cognition-Motivation-Emotional Intelligence-Resistance Skills” (CMER) was developed to enhance cognition upon drug use, to decrease motivation of drug use and to improve emotional adjusting and drug resistance skills in this study. A total of 798 students from 3 senior high schools in Wuhan, a city in central China, were assigned randomly to intervention and control groups. The intervention group received the CMER program in which knowledge, development of positive attitude and motivation towards drugs and training of peer resistance skills were basic elements. The immediate impact was compared by measuring the above mentioned elements prior to and three-month after the training session. Students from both groups were asked to complete a self-administered questionnaire. The questionnaire included demographic items, self-reported drug use behavior, cognition, attitude, and motivation associated with the initiation of drug use and resistance skills. Three months after the intervention, significant effects were found on “illegal substance use at least once” (P<0.05) between the intervention and control groups. Immediate effects of the intervention were also found on knowledge, motivation and peer resistance skills (P<0.05), but there was no clear evidence for any effects on attitude towards substance use (P>0.05). It was concluded that the CMER program, which significantly increased the knowledge of drugs and peer resistance skills, was effective in the drug abuse prevention in a sample of school students in Wuhan, China.

Following acute and chronic liver injury, hepatic stellate cells (HSCs) become activated to undergo a phenotypic transformation into myofibroblast-like cells and lose their retinol content, but the mechanisms of retinoid loss and its potential roles in HSCs activation and liver fibrosis are not understood. The influence of retinoids on HSCs and hepatic fibrosis remains controversial. The purpose of this study was to evaluate the effects of all-trans retinoid acid (ATRA) on cell proliferation, mRNA expression of collagen genes [procollagen α1 (I), procollagen α1 (III)], profibrogenic genes (TGF-β₁, CTGF, MMP-2, TIMP-1, TIMP-2, PAI-1), fibrolytic genes (MMP-3, MMP-13) and the upstream element (JNK and AP-1) in the rat hepatic stellate cell line (CFSC-2G). Cell proliferation was evaluated by measuring BrdU incorporation. The mRNA expression levels of collagen genes [procollagen α1 (I), procollagen α1 (III)], profibrogenic genes (TGF-β₁, CTGF, MMP-2, TIMP-1, TIMP-2, PAI-1), and fibrolytic genes (MMP-3, MMP-13) were quantitatively detected by using real-time PCR. The mRNA expression of JNK and AP-1 was quantified by RT-PCR. The results showed that ATRA inhibited HSCs proliferation and diminished the mRNA expression of collagen genes [procollagen α1 (I), procollagen α1 (III)] and profibrogenic genes (TGF-β₁, CTGF, MMP-2, TIMP-1, TIMP-2, PAI-1), and significantly stimulated the mRNA expression of MMP-3 and MMP-13 in HSCs by suppressing the mRNA expression of JNK and AP-1. These findings suggested that ATRA could inhibit proliferation and collagen production of HSCs via the suppression of active protein-1 and c-Jun N-terminal kinase signal, then decrease the mRNAs expression of profibrogenic genes (TGF-β₁, CTGF, MMP-2, TIMP-1, TIMP-2, PAI-1), and significantly induce the mRNA expression of MMP-3 and MMP-13.

This study was designed to determine the impact of chrysoeriol on proliferation and cell cycle progression in the human multiple myeloma cell lines RPMI 8226 and KM3, and its related molecular mechanisms. Chryseoriol was identified by using the phosphorylated AKT-specific cytoblot high throughput assay. CCK-8 assay was employed to examine the growth inhibition rate and IC₅₀ (48 h) in peripheral blood mononuclear cells (PBMNCs), RPMI 8226 and KM3 cells treated with chrysoeriol at various concentrations. Cells were labeled with 5–6-carboxyfluorescein diacetate succinimidyl ester (CFSE), and the proliferation dynamics was detected by flow cytometry and analyzed with ModFit software. The cell cycles of RPMI 8226 and KM3 cells were measured by flow cytometry when the IC₅₀ concentration of chrysoeriol was adopted. The alterations in cell-cycle related proteins (Cyclin B1, Cyclin D1, p21) and proteins in PI3K-AKT-mTOR pathway were determined by Western blot analysis. The results showed the proliferation of multiple myeloma cells was significantly inhibited by chrysoeriol, resulting in cell cycle arrest in G₂/M phase. Chrysoeriol could significantly reduce the expression of p-AKT (s473) and p-4eBP1 (t37/46) protein, meanwhile enhanced Cyclin B1 and p21 protein expression. Similar effects were not observed in PBMNCs from normal donors. It was concluded that chrysoeriol was a selective PI3K-AKT-mTOR pathway inhibitor. It restrained the proliferation of human multiple myeloma cells, but didn’t affect proliferation of PBMNCs from normal donors. It might exhibit the cell cycle regulatory effect via the inhibition of PI3K-AKT-mTOR signal pathway.

The present study examined the functional profile of dendritic cells (DCs) in patients with rheumatoid arthritis (RA) and the effects of simvastatin on the function of DCs. A total of 40 patients who was recently diagnosed as having RA were equally assigned to two groups: the routine treatment group (group R) and the routine treatment plus simvastatin group (group R+S). Twenty healthy individuals served as control. The peripheral blood mononuclear cells (PBMCs) were isolated before and 4 weeks after the treatment and then cultured with interleukin-4 (IL-4) and granulocyte-macrophage colony stimulatory factor (GM-CSF) to prepare mature DCs. The expression of co-stimulating factor CD86 on the surface of DCs was assessed by flow cytometry. And the stimulating capacity of DCs was measured by mixed lymphocyte reaction (MLR). The contents of cytokines in culture supernatants of DCs in MLR were detected by ELISA. Blood lipids and high-sensitivity C-reactive protein (hs-CRP) were detected. The relationship between the expression of CD86 and the blood CRP level was also investigated. The results showed that, as compared with the control group, the CD86 expression and the level of cytokines secreted by DCs were significantly increased in RA patients and greater stimulating capacity of DCs in MLR was demonstrated in RA patients. T lymphocytes in MLR secreted higher levels of proinflammatory cytokines (IL-2, IL-17, TNF-α and INF-γ) and lower level of anti-inflammation cytokine (IL-10). The function of DCs was markedly weakened and the level of hs-CRP and low-density lipoprotein was substantially lowered in group R+S in comparison to group R. The CD86 expression was positively correlated with hs-CRP. It was concluded that DCs in RA are highly activated and DC-initiated immune reaction may play an important role in the pathogenesis of RA. Simvastatin administration can significantly inhibit the DCs function and reduce the level of hs-CRP, indicating the suppression on inflammatory reaction may be one of the mechanisms by which simvastatin exerts its effect in treating RA.

Neurocytoma, a rare brain tumor, is characterized by a mass located mainly in cerebral ventricles. It is prone to be misdiagnosed as oligodendroglioma or ependymoma due to their similar histopathological features in clinical practice. This study aimed to examine the clinicopathological features and differential diagnosis of central and extraventricular neurocytoma. The clinical and histopathological data of 17 patients (male: female=7:10; age: 4–41 years; mean age: 27.4 years) with central or extraventricular neurocytoma were retrospectively analyzed. These patients showed typical radiological, histopathological and immunohistochemical features of neurocytoma. The tumor tissue was found to be composed of small uniform cells with round nuclei and clear cytoplasm resembling that of oligodendroglioma and ependymoma. Immunohistochemistry revealed the tumor tissues were positive for neuronal markers such as synaptophysin (SYN) and neuronal nuclear antigen (NeuN). It was concluded histopathological features of neurocytoma overlaps with some tumors in the central neural system. Immunopositivity for SYN and NeuN can help differentially diagnose neurocytoma.

In this study, CD133+ subpopulations were isolated from 41 primary colorectal cancer tissues, the proliferation and cell cycle distribution of the cells were examined without in vitro expansion, and then compared to those of cell lines. The detection of CD133 in colorectal cancer tissues, isolation of CD133+ and CD133− epithelial subpopulations, Ki-67/DNA multiparameter assay and cell volume analysis were flow cytometrically conducted. The results showed that Ki-67 expression was correlated with CD133 level in primary cancer tissues, while cell cycle G2/M phase distribution or clinicopathological characteristics was not. In addition, the CD133+ cells showed larger cell volume and higher Ki-67 expression as compared with CD133− cells. But there was no statistically significant difference in G₂/M phase distribution between the two subpopulations. Our results demonstrated that the CD133+ subpopulation in colorectal cancer tissue contained more actively cycling and proliferating cells, which was not correlated to clinicopathological factors but might contribute to tumor progression and poor clinical outcome.

This study investigated the effect of TNP-470 in combination with carmustine (BCNU) on the growth of subcutaneously implanted human glioblastoma xenografts in nude mice. Human glioblastoma U-251 cells (1×10⁷) were injected into 24 nude mice subcutaneously. The tumor-bearing mice were randomly divided into 4 groups on the seventh day following tumor implantation: TNP-470 group, in which TNP-470 was given 30 mg/kg subcutaneously every other day 7 times; BCNU group, in which 20 mg/kg BCNU were injected into peritoneal cavity per 4 days 3 times; TNP-470 plus BCNU group, in which TNP-470 and BCNU were coadministered in the same manner as in the TNP-470 group and the BCNU group; control group, in which the mice were given 0.2 mL of the mixture including 3% ethanol, 5% acacia and 0.9% saline subcutaneously every other day 7 times. The tumor size and weights were measured. The tumor microvessel density (MVD) was determined by immunostaining by using goat-anti-mouse polyclonal antibody CD105. The results showed that on the 21th day following treatment, the volume of xenografts in the TNP-470 plus BCNU group was (108.93±17.63)mm³, markedly lower than that in the TNP-470 group [(576.10±114.29)mm³] and the BCNU group [(473.01±48.04)mm³] (both P<0.01). And the xenograft volume in these 3 treatment groups was even much lower than that in the control group [(1512.61±470.25) mm³] (all P<0.01). There was no significant difference in the volume of xenografts between the TNP-470 group and the BCNU group (P>0.05). The inhibition rate of the tumor growth in the TNP-470 plus BCNU group was (92.80±11.37)%, notably higher than that in the TNP-470 group [(61.91±6.29)%] and the BCNU group [(68.73±9.65)%] (both P<0.01) on the 21th day following treatment. There was no significant difference in the inhibition rate of tumor growth between the TNP-470 group and the BCNU group (P>0.05). The MVD of xenografts in the TNP-470 plus BCNU group was decreased significantly as compared with that in the TNP-470 group or the BCNU group (both P<0.05). The MVD of xenografts in the 3 treatment groups was markedly reduced as compared with that in the control group (all P<0.05). No significant changes in weights were observed before and after the treatment in each group (all P>0.05). It was concluded that the combination of TNP-470 and BCNU can significantly inhibit the growth of human glioblastoma xenografts in nude mice without evident side effects.

To explore the clinical classification of hamate hook fracture and the treatment strategy for different type of fractures, 12 patients who suffered from hamate hook fractures were followed up retrospectively. According to the fracture sites and the prognosis, we classified the hamate hook fractures into 3 types. Type I referred to an avulsion fracture at the tip of hamate hook, type II was a fracture in the middle part of hamate hook, and type III represented a fracture at the base of hamate hook. By the classification, in our series, only 1 fell into type I, 7 type II, and 4 type III. The results were evaluated with respect to the functional recovery, recovery time and the association among the clinical classification, pre-operative complications and treatment results. The average follow-up time of this group was 8.4±3.9 months. Two cases were found to have fracture non-union and both of them were type II fractures. Six patients had complications before operation. Five cases were type II fractures and 1 case type III fracture. All the patients were satisfied with the results at the time of the last follow-up. Their pain scale and grip strength improved significantly after treatment. All the pre-operative complications were relieved. The recovery time of hamate hook excision was significantly shorter than that of the other two treatments. The incidences of both pre-operative complications and non-union in type II fractures were higher than those in type I and type III fractures. It was concluded that, generally, the treatment effects with hamate hook fracture are quite good. The complication incidence and prognosis of the fracture are closely related to the clinical classification. Early intervention is critical for type II fractures.

This study examined in vitro effect of lipoxin A₄ (LXA₄) on interleukin-1β (IL-1β) production of monocytes and its possible mechanism in severe preeclampsia (PE). Peripheral venous blood was drawn from 15 patients with severe preeclampsia (PE group) and 20 normal pregnant women (control group) to prepare monocytes which were then treated with LXA₄ at different concentrations of 0, 10, 100 nmol/L respectively. IL-1β level in the supernatant of monocytes was detected by enzyme linked immunoassay. The [Ca²⁺]_i of monocytes was measured by laser scanning confocal microscopy. The results showed that the IL-1β level and the [Ca²⁺]_i of monocytes in the PE group were significantly higher than those in the control group. LXA₄ significantly decreased the generation of IL-1β in a dose-dependent manner in the PE group. After treatment with 100-nmol/L LXA₄, in the PE group, the [Ca²⁺]_i concentration of monocytes was significantly reduced. It was concluded that LXA₄ may inhibit the IL-1β production of monocytes from severe preeclampsia women by inhibiting extracellular calcium influx.

The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation, differentiation, invasion and metastasis of cancer cells, but its role in cervical cancer is still unclear. This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors. The NDRG1 expression in cervical tissues and cells was detected by RT-PCR. Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines. The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting, respectively. Its effects on cell proliferation, migration, invasion, cell cycle and apoptosis were detected by MTT, transwell migration assay and flow cytometry (FCM), respectively. The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001). After transfection with pEGFP-N1-NDRG1-GFP, the mRNA and protein expression of NDRG1 was up-regulated in Siha cells, which suppressed cell proliferation (P<0.001), induced cell cycle arrest (P<0.05), reduced invasion and migration of Siha cells (P<0.05), but caused no cell apoptosis. Moreover, vascular endothelial growth factor (VEGF), a tumor-induced angiogenesis factor, was markedly reduced and E-cadherin, a cell adhesion molecule, was increased in the cells transfected with pEGFP-N1-NDRG1-GFP. It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth, invasion and metastasis of human cervical cancer.

The present study examined von Willebrand factor (vWF) levels and ADAMTS13 activity in pregnant and severe preeclamptic women in order to shed light on the prothrombotic state in severe preeclampsia. Thirty healthy women of childbearing age, 22 second trimester pregnant women, 30 third trimester pregnant women and 10 severe preeclamptic patients were recruited in this study. ADAMTS13 activity was determined by the FRETS-vWF73 assay and vWF antigen (vWF:Ag) levels by an enzyme-linked immunosorbent assay. The results showed that there were statistically significant differences in plasma vWF antigen levels between the severe preeclamptic and third trimester pregnant women, between third and second trimester pregnant women (P<0.05). The third trimester pregnant women had significantly lower plasma ADAMTS13 activity than second trimester pregnant women (P<0.05). Nevertheless, no significant differences in plasma ADAMTS13 activity were found between severe preeclamptic patients and the third trimester pregnant women (P>0.05). In conclusion, plasma ADAMTS13 activity is normal in severe preeclampsia despite the increased vWF:Ag levels. Prothrombotic state is involved in the pathogenesis of severe preeclampsia, as a result of endothelial injury.

Brain-derived neurotrophic factor (BDNF) can promote developmental competence in mammalian oocytes during in vitro maturation (IVM), but the role of BDNF in oocyte maturation at cellular level is not still clear. In this study, mouse cumulus-enclosed oocytes subjected to IVM were fertilized and cultured to blastocyst stage. Meiotic spindle configuration and cortical granules distribution during oocyte maturation in vitro were assessed by using immunofluorescence and laser confocal microscopy. The results showed that BDNF contributed to the complete preimplantation development of mouse oocytes compared to the control oocytes (13.78% vs. 5.92%; P<0.05). Further, BDNF did not accelerate nuclear maturation of IVM oocytes. For the BDNF-treated oocytes at meiosis I, Meiotic spindle areas were significantly smaller and the number of cytoplasmic microtubule organizing centers was greater than that in the control, and the percentages of oocytes showed spindles positioned near the oolemma and a well-formed cortical granule-free domain were significantly higher than that of the control. These morphological characteristics of the BDNF-treated oocytes were much closer to the oocytes matured in vivo than those of the control oocytes. In conclusion, BDNF can promote the developmental competence of mouse IVM oocytes, by improving the meiotic spindle configuration and location and cortical granules distribution at meiosis.

Dynamic color is an important carrier that takes information in some special occupations. However, up to the present, there are no available and objective tests to evaluate dynamic color processing. To investigate the characteristics of dynamic color processing, we adopted two patterns of visual stimulus called “onset-offset” which reflected static color stimuli and “sustained moving” without abrupt mode which reflected dynamic color stimuli to evoke event-related brain potentials (ERPs) in primary color amblyopia patients (abnormal group) and subjects with normal color recognition ability (normal group). ERPs were recorded by Neuroscan system. The results showed that in the normal group, ERPs in response to the dynamic red stimulus showed frontal positive amplitudes with a latency of about 180 ms, a negative peak at about 240 ms and a peak latency of the late positive potential (LPP) in a time window between 290 and 580 ms. In the abnormal group, ERPs in response to the dynamic red stimulus were fully lost and characterized by vanished amplitudes between 0 and 800 ms. No significant difference was noted in ERPs in response to the dynamic green and blue stimulus between the two groups (P>0.05). ERPs of the two groups in response to the static red, green and blue stimulus were not much different, showing a transient negative peak at about 170 ms and a peak latency of LPP in a time window between 350 and 650 ms. Our results first revealed that some subjects who were not identified as color blindness under static color recognition could not completely apperceive a sort of dynamic red stimulus by ERPs, which was called “dynamic red blindness”. Furthermore, these results also indicated that low-frequency ERPs induced by “sustained moving” may be a good and new method to test dynamic color perception competence.

Congenital cataract is a highly heterogeneous disorder at both the genetic and the clinical-phenotypic levels. A unique cataract was observed in a 4-generation Chinese family, which was characterized by autosomal dominant inheritance and late-onset. Mutations in the 13 known genes (CRYAA, CRYAB, CRYBB1, CRYBB2, CRYGC, CRYBA1/A3, CRYGD, Connexin50, Connexin46, intrinsic membrane protein LIM2, cytoskeletal protein BFSP2, the major intrinsic protein-MIP and the heat shock factor HSF4) have previously been demonstrated to be the frequent reason for isolated congenital cataracts, but the exact molecular basis and underlying mechanisms of congenital cataract still remain unclear. This study was designed to find whether these 13 genes developed any mutation in the family members and to identify the disease-causing gene. Polymerase chain reaction (PCR) and direct DNA sequence analysis were carried out to detect the 13 genes. The results showed that no mutation causing amino acid alternations was found in these potential candidate genes among all patients in the family, and only several single-nucleotide polymorphisms (SNPs) were identified. A transitional mutation in the fourth intron of CRYBB2 and some silent mutations in the first exon of BFSP2 and CRYGD were found in the cataract family, but further study showed that these mutations could also be found in normal controls. It was concluded that some unidentified genes may underlie the occurrence of late-onset cataract in this family. A genome-wide screening will be carried out in the next study.

Pigment epithelium derived factor (PEDF) has been proven to be an effective drug for the treatment of choroidal neovascularization (CNV). However, the lack of ideal administration route is the biggest bottleneck preventing PEDF from wider clinical use. In this study, we developed a novel PEDF-carrying system which employed immuno-nano-liposomes (INLs) under ultrasound exposure. PEDF-loaded INLs were prepared by conjugating nanoliposomes to the peptide ATWLPPR specifically targeting the receptor-2 for vascular endothelial growth factor (VEGFR-2) and reversely encapsuling PEDF. RF/6A cells were incubated with PEDF-loaded INLs. CNV models of BN rats were injected with PEDF-loaded INLs. MTT assay was used to evaluate the cytotoxicity of the INLs on RF/6A cells. Flow cytometry was conducted to detect the apoptotic rate of cells. Laser scanning confocal microscopy was employed to observe the binding and transmitting process of PEDF-loaded INLs and to calculate the area of CNV in the rat model. The results showed that the PEDF-loaded INLs could exclusively bind to CNV but not to the normal choroidal vessels. The CNV area was significantly decreased in PEDF treatment groups in comparison with control group (P<0.05). Moreover, PEDF-loaded INLs exposed under ultrasound were more efficient in reducing the CNV area (P<0.05). It was concluded that INLs in combination with ultrasonic exposure can transmit PEDF into cytoplasma with high specificity and efficiency, which strengthens the inhibitory effects of PEDF on CNV and reduces its side effects. PEDF-loaded INLs possibly represent a new treatment paradigm for patients with ocular neovascularization.

This study examined the corneal permeability of topical eye drop solutions added with various corneal penetrating accelerators and gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) by nuclear magnetic resonance imaging (MRI). Twenty-four New Zealand rabbits were randomly divided into 3 groups according to the random digits table: Gd-DTPA group, in which the rabbits received 23.45% Gd-DTPA; hyaluronic acid group, in which 23.45% Gd-DTPA plus 0.2% hyaluronic acid was administered; azone group, in which 23.45% Gd-DTPA with 0.2% azone was given. Fifty microliters of the eye drops was instilled into the conjunctive sac every 5 min, for a total of 6 applications in each group. Contrast medium signals in the cornea, anterior chamber, posterior chamber, and vitreous body were scanned successively by MRI. The morphology and cell density of the corneal endothelium were examined before and 24 h after the treatment. The results showed that the residence time of Gd-DTPA in the conjunctival sac in the hyaluronic acid and azone groups was longer than that in the Gd-DTPA group. The signals in the anterior chamber of the Gd-DTPA and hyaluronic acid groups were increased slightly, and those in the azone group strengthened sharply. The signal intensity continuously rose over 80 min before reaching plateau. The strengthening rate of signals in the anterior chamber was 19.63% in the Gd-DTPA group, 53.42% in the sodium hyaluronate group, and 226.94% in the azone group. No signal was detected in the posterior chamber or vitreous body in all the 3 groups. Corneal morphology and cell density did not show any significant changes after the treatment in all the 3 groups. It was concluded that azone can significantly improve the corneal permeability of drugs that are similar to Gd-DTPA in molecular weight and molecular size, and MRI is a noninvasive technique that can dynamically detect eye drop metabolism in real time.

The effects of DK2, a peroxisome proliferator-activated receptor γ agonist, on cultured human pterygium fibroblasts (HPFs) in virto were studied. The HPFs were incubated with 0–200 μmol/L DK2 for 12–72 h. The MTT method was used to assay the bio-activity of DK2 at different doses and time. The cytotoxic effect of DK2 was measured by LDH release assay. The cell cycle distribution and apoptosis were flow cytometrically detected. The expression of proliferating cell nuclear antigen (PCNA) in each group was detected by real-time PCR (RT-PCR) and Western blotting. The results showed that administration of 1–75 μmol/L DK2 for 12–72 h could significantly inhibit HPF proliferation in a dose- and time-dependent manner. DK2-treated cells did not release significant amount of LDH as compared with rosiglitazone-treated cells. After treatment with DK2 at concentrations of 15, 25 μmol/L for 24 h, the number of HPFs in G₀/G₁ phase was significantly increased while that in S phase was significantly decreased (P<0.05), leading to arrest at G₀/G₁ phase. The apoptosis rates of HPF cells in drug-treated groups were significantly higher than the rate of control group (P<0.05). At the dosage range between 15–25 μmol/L, DK2 could inhibit the expression of PCNA mRNA and protein in HPFs in a dose-dependent fashion (P<0.05). It was concluded that PPARγ agonist can significantly inhibit HPF proliferation, resulting in the arrest at G₀/G₁ phase, induce the apoptosis of HPFs, and suppress the synthesis of PCNA, in dose- and time-dependent manners.

The study examined the association between calcified liver metastases and the histopathology of the primary colorectal carcinoma in Chinese. The clinical, pathological and CT data were retrospectively analyzed in 210 patients (mean age: 54.2 years) with liver metastases from colorectal carcinoma. Plain CT scanning and contrast-enhanced scanning were performed in all the patients. For the contrast-enhanced examination, iohexol was injected by using a high pressure syringe at a flow rate of 2.5–3.0 mL/s. The arterial phase lasted approximately 25 s and the portal venous phase about 60 s. All patients had no history of chronic liver diseases and had never received interventional treatments. χ²-test was used to analyze the rate of calcification in the liver metastasis from colorectal cancer of different differentiation degrees. Among the 210 cases of liver metastases, 22 patients (10.5%) were found to have calcified liver metastases on CT scan. Two patients with calcified liver metastasis received lumpectomy and developed calcification in recurrent tumors. Another two patients had calcification in newly developed tumor masses. And the calcification in the newly developed masses was similar to that of their primary counterparts in terms of morphology and distribution. On the enhanced CT scan, the tumors exhibited no enhancement during hepatic arterial phase and showed slight rim enhancement during portal venous scan in the 22 cases. The calcification became obscure on contrast-enhanced scans. Histopathologically, the primary tumors were well-differentiated adenocarcinoma in 6 cases, moderately-differentiated adenocarcinoma in 10, poorly-differentiated adenocarcinoma in 4 and mucinous adenocarcinoma in 2 among the 22 cases. No statistical correlation was noted between the incidence of calcified liver metastasis and the pathological subtypes and differentiation degrees of the primary colorectal carcinoma. It was concluded that calcified liver metastases may result from colorectal adenocarcinomata of different differentiation degrees or mucinous adenocarcinomata in Chinese population. There is no correlation between calcification of liver metastases and the pathological subtype of the primary colorectal carcinoma in Chinese, which is different from the findings that calcified metastases were associated with colorectal mucinous adenocarcinoma in other ethnic groups.

The value of the left atrial volume tracking (LAVT) method in the evaluation of left atrial (LA) function in patients with diabetes mellitus (DM) was examined in this study. Fifty-eight DM patients as DM group and 40 healthy people as normal control group were enrolled in this study. EUB-6500 echocardiographic imaging system with LAVT was applied to display and analyze the LA volume curve imaging on LV apical two and four chamber views. The maximal LA volume at end-systole (LAV_max), LA volume at the onset of ECG-P wave (LAV_p), the minimal LA volume at end-diastole (LAV_min) from the LA volume curve were acquired and recorded. All values above were standardized by body surface area (BSA). Then the passive, active and total LA volume (LAVIpass, LAVIact, LAVItotal) and empting rate (%LAVIpass, %LAVIact, %LAVItotal), effective passive and active empting rate (%eLAVIpass, %eLAVIact), and the proportionality of passive empting volume and active empting volume were calculated. The LAVIp, LAVIact, LAVItotal, %LAVIact, %LAVItotal and %eLAVIact were significantly higher in the DM group than those in the control group, whereas the LAVIpass, %LAVIpass, %eLAVIpass and LAVIpass/act were lower (all P<0.05). For the LA volume change in DM, the active empting volume was enhanced at end-diastole. It was concluded that LAVT is a potentially useful tool to evaluate the function of LA.

We report a case of reversible hepatofugal portal flow after auxiliary partial orthotopic liver transplantation (APOLT) from a living donor in this study. On postoperative day 6, continuous hepatofugal portal flow was observed in the grafted liver without portal thrombosis and obstruction of the hepatic vein. Based on histological findings, acute rejection was the suspected cause. The normal portal venous flow was restored after steroid pulse and antithymocyte globulin (ATG) therapies. The patient was discharged on the 30th postoperative day. It was concluded that hepatofugal flow after liver transplantation is a sign of serious acute rejection, and can be successfully treated by anti-rejection therapy.