This study explored the molecular mechanisms underlying the time-dependent autophagy and apoptosis induced by nutrient depletion in human multiple myeloma cell line RPMI8226 cells. RT-PCR and qRT-PCR were used to evaluate the transcriptional levels of Deptor, JNK1, JNK2, JNK3, Raf-1, p53, p21 and NFκB1 at 0, 6, 12, 18, 24 and 48 h after nutrient depletion in RPMI8226 cells. We found that transcriptional levels of Deptor were increased time-dependently at 0, 6, 12 and 18 h, and then decreased. Its alternation was consistent with autophagy. Transcriptional levels of Raf-1, JNK1, JNK2, p53 and p21 were increased time-dependently at 0, 6, 12, 18, 24 and 48 h accompanying with the increase of apoptosis. Transcriptional levels of NFκB1 at 6, 12, 18, 24 and 48 h were decreased as compared with 0 h. It was suggested that all the studied signaling molecules were involved in cellular response to nutrient depletion in RPMI8226 cells. Deptor contributed to autophagy in this process. Raf-1/JNK /p53/p21 pathway may be involved in apoptosis, and NFκB1 may play a possible role in inhibiting apoptosis. It remained to be studied whether Deptor was involved in both autophagy and apoptosis.

T cell immunoglobulin mucin (TIM) family plays a key role in regulating immune responses. In this study, the interactions of human TIM family with apoptotic cells were evaluated in order to provide a foundation for further study on the roles of human TIM genes in apoptosis. Nine kinds of pEGFP-N1 eukaryotic expression vectors containing different lengths of the three members of human TIM genes for the expression of TIM-EGFP and the vectors for the expression of TIM-Fc fusion proteins were constructed. It was found that human TIM proteins could recognize and bind to apoptotic cells directly, but not to viable cells. The interactions of sTIM-1-EGFP, sTIM-3-EGFP and sTIM-4-EGFP with apoptotic cells were blocked by TIM-1-Ig, TIM-3-Ig and TIM-4-Ig fusion proteins respectively. In addition, human TIM proteins mediated the recognition of apoptotic cells and bound to apoptotic cells directly via the IgV domains. In conclusion, the TIM family may play a key role in the regulation of apoptosis. Our data also suggest that human TIM proteins probably serve as novel proteins for the detection of the early cellular apoptosis.

Pim kinases contribute to tumor formation and development of lymphoma, which shows enhanced DNA replication, DNA recombination and repair. Endothelial cells^(ECs) express all the three members of Pim kinase gene family. We hypothesized that DNA repair gene would regulate Pim expression in ECs. Human umbilical vein endothelial cells (HUVECs) were isolated and maintained in M199 culture medium. The cellular distribution of Pim-3 in ECs was determined by immunofluorescent staining. The siRNA fragments were synthesized and transfected by using Lipofectamine LTX. The total cellular RNA was extracted from the cells by using Trizol reagent. cDNAs were quantified by semi-quantity PCR. The effects of LY294002 and wortmannin on RNA stability in ECs were also examined. Our data showed that LY294002 and wortmannin, phosphatidylinositol 3-kinase (PI3K) and PI3K-like kinase inhibitors, increased Pim mRNA expression in ECs without altering the mRNA stability. RNA interference (RNAi) targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) increased mRNA expression of Pim-3 and Pim-1, respectively. Silencing of Akt decreased Pim-1 instead of Pm-2 and Pim-3 gene expression in ECs. But etoposide, a nucleoside analogue, which could activate DNA-PKcs and ATM, increased Pim expression in ECs. Our study indicates that the expression of Pim kinases is physiologically related to DNA-PKcs and ATM in ECs.

Mounting evidence supports that a newly identified regulatory T cell (Treg), CD4⁺LAP⁺ Treg, is associated with oral tolerance induction and following inhibition of atherosclerosis, but little is described about whether nasal tolerance to antigen likewise induces the novel Tregs production and the relevant antiatherosclerotic benefit. We investigated the effect of nasal administration of heat shock protein-60 (HSP60) on atherogenesis. HSP60 or phosphate buffer solution (PBS) was nasally administered to six-week-old male ApoE^−/− mice. At the 10th week after the nasal administration, there was a significant decrease in atherosclerotic plaque areas of aortic roots in the HSP60-treated mice as compared with those in the PBS-treated mice. Atherosclerosis suppression was accompanied with a significant increase in CD4⁺LAP⁺ and CD4⁺CD25⁺Foxp3⁺ Tregs and a concurrently increased production of TGF-β in the HSP60-treated mice. The protective effect of HSP60 was offset by injection of anti-TGF-β antibody. It is concluded that nasal administration of HSP60 can inhibit atherosclerotic formation through immune tolerance which is established by Tregs depending on the induction of anti-inflammatory cytokine TGF-β. Immune tolerance induced by nasal administration of HSP60 may provide an alternative therapeutic method for atherosclerosis.

Angiotensin II (ANGII) plays an important role in the pathogenesis of atherosclerosis by inducing proliferation of vascular smooth muscle cells (VSMCs). In our study, we observed the effects of valsartan on proliferation of cultured VSMCs treated with or without ANGII by cell counting and methyl thiazolyl tetrazolium (MTT) assay, and detected the expression of mitofusin 2 (Mfn2), a newly discovered cell proliferation inhibitor and a related cell proliferation signaling pathway protein by Western blotting. ANGII at a concentration of 10⁻⁶ mol/L significantly stimulated VSMCs proliferation, down-regulated the expression of Mfn2 and up-regulated the expression of Raf and ERK1/2. Valsartan inhibited such effects of ANGII at concentrations of 10⁻⁵ and 10⁻⁶ mol/L, but not at 10⁻⁷ mol/L. Valsartan had no significant effect on the proliferation of untreated VSMCs. These results suggest that valsartan inhibits ANGII-induced proliferation of VSMCs in vitro via Mfn2-Ras-Raf-ERK/MAPK signaling pathway.

Bone marrow mesenchymal stem cells (MSCs) have shown potential for cardiac repair following myocardial injury, but this approach is limited by their poor viability after transplantation. The present study was to investigate whether trimetazidine (TMZ) could improve survival of MSCs in an ex vitro model of hypoxia, as well as survival, differentiation, and subsequent activities of transplanted MSCs in rat hearts with acute myocardial infarction (AMI). MSCs at passage 3 were examined for their viability and apoptosis under a transmission electron microscope, and by using flow cytometry following culture in serum-free medium and exposure to hypoxia (5% CO₂, 95% N₂) for 12 h with or without TMZ. Thirty Wistar rats were divided into 3 groups (n=10 each group), including group I (AMI control), group II (MSCs transplantation alone), and group III (TMZ+MSCs). Rat MSCs (4×10⁷) were injected into peri-infarct myocardium (MSCs group and TMZ+MSCs group) 30 min after coronary artery ligation. The rats in TMZ+MSCs group were additionally fed on TMZ (2.08 mg·kg⁻¹·day⁻¹) from day 3 before AMI to day 28 after AMI. Cardiac structure and function were assessed by echocardiography at 28th day after transplantation. Blood samples were collected before the start of TMZ therapy (baseline), and 24 and 48 h after AMI, and inflammatory cytokines (CRP, TNF-α) were measured. Then the survival and differentiation of transplanted cells in vivo were detected by immunofluorescent staining. The cellular apoptosis in the peri-infarct region was detected by using TUNEL assay. Furthermore, apoptosis-related proteins (Bcl-2, Bax) within the post-infarcted myocardium were detected by using Western blotting. In hypoxic culture, the TMZ-treated MSCs displayed a two-fold decrease in apoptosis under serum-free medium and hypoxia environment. In vivo, cardiac infarct size was significantly reduced, and cardiac function significantly improved in MSCs and TMZ+MSCs groups as compared with those in the AMI control group. Combined treatment of TMZ with MSCs implantation demonstrated further decreased MSCs apoptosis, further increased MSCs viability, further decreased infarct size, and further improved cardiac function as compared with MSCs alone. The baseline levels of inflammatory cytokines (CRP, TNF-α) had no significant difference among the groups. In contrast, all parameters at 24 h were lower in TMZ+MSCs group than those in MSCs group. Furthermore, Western blotting indicated that the expression of anti-apoptotic protein Bcl-2 was up-regulated, while the pro-apoptotic protein Bax was down-regulated in the TMZ+MSCs group, compared with that in the MSCs group. It is suggested that implantation of MSCs combined with TMZ treatment is superior to MSCs monotherapy for MSCs viability and cardiac function recovery.

The effects of testosterone on norepinephrine release were investigated in the isolated rat hearts. Sprague-Dawley male rats (n=120) were randomized to testosterone and control groups. The rats in testosterone group were perfused with modified Krebs-Henseleit buffer containing different concentrations of testosterone (0.1, 1.0, 10.0, and 100.0 nmol/L, respectively). Myocardial ischemia was induced by globally stopping the perfusion flow. Exocytotic norepinephrine release was induced by electrical field stimulation at 5 V (effective voltage) and 6 Hz (pulse width of 2 ms) for 1 min. The overflow of norepinephrine was determined by high pressure liquid chromatography and electrochemical detection (HPLC-EC). Following acute ischemia, testosterone (1.0, 10.0 and 100.0 nmol/L) significantly reduced norepinephrine release (P<0.01), and the norepinepherine overflow was similar between the control and 0.1 nmol/L testosterone group (P>0.05). Electrical stimulation of the ventricle evoked norepinepherine release, and this was diminished by the perfusion with testosterone at the concentrations of 1.0, 10.0 and 100.0 nmol/L (P<0.01). It is suggested that testosterone suppresses ischemia- and electrical stimulation-induced norepinepherine release in the isolated rat hearts.

To explore the relationship between serum thyroid stimulating hormone (TSH) level and obesity and nonalcoholic fatty liver disease (NAFLD) in euthyroid subjects, 1322 subjects were subjected to a questionnaire survey and physical examination. Fasting blood samples were collected to test serum TSH, plasma glucose and lipids. Fatty liver was diagnosed by type B ultrasonography. The relationship between serum TSH level and body mass index (BMI), percentage of body fat and NAFLD was analyzed. The results showed that serum TSH level was significantly higher in females than in males at the same group, and it was significantly higher in overweight group than in control group. Levels of body weight, BMI, waist circumference and percentage of body fat were increased in TSH >2.5 group compared to TSH ≤2.5 group in women. However, plasma lipids showed no significant differences. In males all the parameters showed no significant differences between two groups. Serum TSH was significantly correlated with body weight, BMI, waist circumference and percentage of body fat after adjustment for age in females. Multiple linear regression analysis revealed that percentage of body fat and BMI contributed significantly to the variance of TSH. Serum TSH level was significantly higher in nonalcoholic fatty liver group than in normal group in females. Multiple logistic regression analysis showed that TSH level was not the independent risk factor of NAFLD. Taken together the data suggest that serum TSH in normal range is significantly correlated with BMI and percentage of body fat in females. And the change of TSH level would not influence the prevalence of NAFLD.

The molecular mechanism by which obesity induces insulin resistance is not completely understood. The aim of this study was to determine how lipopolysaccharide-induced tumor necrosis-α factor (LITAF) influenced obesity-induced insulin resistance using a cellular co-culture system. The cells were divided into 3 groups: palmitic acid (PA) stimulation group, LITAF small interfering RNA (siRNA) group and untreated (NC) group. The LITAF siRNA was used for knockdown of LITAF expression in human THP-1 macrophages. The expression levels of LITAF, IRS-2, IRS-2Tyr465, PI3K, and GLUT2 in each group were measured by using quantitative reverse transcriptase real-time polymerase chain reaction and Western blotting. The expression of LITAF was much higher in the PA group than in the siRNA and NC groups (*P<0.05); meanwhile, the expression of IRS-2, IRS-2Tyr465, PI3K, and GLUT2 was much lower in the PA group than in the NC group (*P<0.05); however, IRS-2, IRS-2Tyr465, PI3K, and GLUT2 had much higher expression in the siRNA group than in the PA group (*P<0.05). It is concluded that PA can induce insulin resistance in liver cells and knockdown of LITAF expression can reduce insulin resistance in liver cells, suggesting LITAF may regulate the insulin signal transduction pathway involved in obesity-induced insulin resistance.

In this study, the effects of hyperosmolality on the expression of urea transporter A2 (UTA2) and aquaporin 2 (AQP2) were investigated in transfected immortalized mouse medullary collecting duct (mIMCD3) cell line. AQP2-GFP-pCMV6 and UTA2-GFP-pCMV6 plasmids were stably transfected into mIMCD3 cells respectively. Transfected mIMCD3 and control cells were cultured in different hypertonic media, which were made by NaCl alone, urea alone, or an equiosmolar mixture of NaCl and urea. The mRNA and protein expression of AQP2 was elevated by the stimulation of NaCl alone, urea alone and NaCl plus urea in AQP2-mIMCD3 cells; whereas NaCl alone and NaCl plus urea rather than urea alone increased the mRNA and protein expression of UTA2 in UTA2-mIMCD3 cells, and all the expression presented an osmolality-dependent manner. Moreover, the mRNA and protein expression of UTA2 rather than AQP2 was found to be synergistically up-regulated by a combination of NaCl and urea in mIMCD3 cells. It is concluded that NaCl and urea synergistically induce the expression of UTA2 rather than AQP2 in mIMCD3 cells, and hyperosmolality probably mediates the expression of AQP2 and UTA2 through different mechanisms.

In order to analyze the cai]ses of delayed diagnosis and raise the level of early diagnosis of atypical multiple myeloma (MM), the differences of presenting features between the patients presented to nephrologists and those presented to hematologists were compared. MM patients in our hospital were studied retrospectively. Those who referred renal impairment were divided into two groups: group I presented to nephrologists prior to MM diagnosis (n=29) and group II presented to hematologists directly (n=62). The age, sex, initial symptoms, haematological and biochemical parameters, the phenotype of paraprotein, bone marrow biopsy and cytology were undertaken and analyzed. The results showed that the median time between the initial symptoms and diagnosis in the patients of group I was longer than that in group II (P<0.001); patients in group I had significantly lower incidence of bone pain (P<0.01) and worse renal function (P<0.05) on presentation. There were lower level of myeloma cells (P<0.05), lower incidence of hypergammaglobulinemia (P<0.05), lower positive rate of monoclonal immunoglobulin (M protein) (P<0.05) and M protein level (P<0.05) in the patients of group I than those in group II. The ratio of monoclonal to lambda monoclonal proteins in a population was 1:3.67 in patients of group I, whereas 1:0.90 in patients of group II (P<0.01). Moreover, patients with λ type had a higher degree of renal insufficiency than those with κ type (P<0.05). It was suggested that the median time between the initial symptoms and diagnosis in the patients presented to nephrologists was longer than that in those presented to hematologists; the patients presented to nephrologists had the lower incidence of bone pain, lower level of myeloma cells and M protein, which made early diagnosis more difficult; more patients presented to nephrologists had the majority of λ light chain type, moreover, patients with λ light chain type had a higher incidence of renal insufficiency.

The noninvasive measurement of liver stiffness (LS) was evaluated by transient elastography (FibroScan) and the possible influencing factors from the patients’ clinical situations including age, gender, liver inflammation represented by alanine transaminase (ALT) and total billirubin (TBIL) level, HBV replication (HBV DNA loads), portal vein pressure (portal vessel diameter, PVD), splenic thickness (SPT) and body mass index (BMI) were analyzed in patients with chronic hepatitis B (CHB). A total of 466 patients including 31 patients with acute-on-chronic liver failure (ACLF), and 435 patients with chronic hepatitis B (CHB) among which 82 patients were diagnosed with liver cirrhosis (LC) by clinical manifestations and liver B-type ultrasonic inspection were enrolled at Tongji Hospital from April to December 2009. LS was measured by a FibroScan device (EchoSens, France). Simultaneously, ALT and TBIL levels, HBV DNA loads, PVD, SPT and BMI in all patients were also tested. Forty-one healthy volunteers served as controls. The values of LS were correlated positively with ages of CHB patients and significantly higher in males than in females. In patients with BMI>28 kg/m² (obesity) and abnormal levels of ALT and TBIL, LS values were significantly increased as compared with those having normal levels of ALT and TBIL. The patients with ACLF had the highest LS value. Furthermore, LS values in the patients with LC were significantly higher than those in patients without LC. It is concluded that noninvasive measurement of liver fibrosis by FibroScan provides an alternative method to evaluate liver fibrosis of patients with CHB. In order to properly illustrate the stiffness value taken by transient elastography, patients’ gender should be taken into consideration and it is also suggested to avoid possible influencing factors including liver inflammation (high levels of ALT and TBIL) and obesity (high BMI).

The osteogenic in vitro effect of low intensity pulsed ultrasound (LIPUS) on SD rat adipose-derived stem cells (ADSCs) was investigated. Rat ADSCs underwent LIPUS (intensity=100 mW/cm²) or sham exposure for 8 min per treatment once everyday in vitro, and then the alkaline phosphatase (ALP) activity and mineralized nodule formation were assessed to evaluate the osteogenic effect of LIPUS on ADSCs. To further explore the underlying mechanism, the osteogenic-related gene mRNA expression was determined by using reverse transcriptase-polymerase chain reaction (RT-PCR) at 1st, 3rd, 5th, 7th day after exposure repectively. Westen blot was used to evaluate the protein expression levels of two osteogenic differentiation associated genes at 7th and 14th day repectively. It was found that ALP activity was increased after LIPUS exposure and LIPUS resulted in mineralized nodule formation of ADSCs in vitro. LIPUS-treated ADSCs displayed higher mRNA expression levels of runt-related transcription factor 2 (Runx2), osteocalcin (OCN), ALP and bone sialoprotein (BSP) genes than controls, and the protein levels of Runx2 and BSP were also increased. The results suggested that LIPUS may induce the osteogenic differentiation of ADSCs in vitro.

Matrix metalloproteinase-2 (MMP-2) level and the ERK1/2 signal pathway are dependent factors for the growth and metastasis of cancer. However, the impact of MMP-2 in combination with ERK1/2 in tumor patients with drug resistance is unknown. To determine the relationship between MMP-2 and the ERK1/2 signal pathway, we established an adriamycin (ADM)-induced MG-63 (ADM-MG-63) cell line. With the increase of the ERK1/2 pathway blocker PD98059, we detected the expression levels of MMP-2 and p-ERK1/2 by Western blot in ADM-MG-63 cells. In ADM-MG-63 cells transfected with MMP-2-siRNA, the expression of ERK1/2 was detected for understanding the function of the ERK1/2 signal pathway. Three siRNAs for MMP-2 (MMP-2-siRNA) were designed, and the optimal one was selected and tested at different time points of 24, 48 and 72 h. Under an ADM-induced condition, ADM-MG-63 cells were finally stable living in the medium of ADM (200 ng/mL). PD98059 could effectively suppress the expression levels of p-ERK1/2 and MMP-2. When the MMP-2 was silenced by using MMP-2-siRNA, the expression of p-ERK1/2 was enhanced. It is concluded that MMP-2 may be involved in ADM resistance dependent on ERK1/2 signal pathway, suggesting interference in ERK1/2 may be a new method of targeted therapy for tumor resistance.

Bone marrow mesenchymal stem cells (BMSCs) have been shown to be multipotent cells that possess high self-replicating capacity. The purpose of our study was to investigate the feasibility of using enteric neuron-like cells obtained by in vitro induction and differentiated from rat BMSCs for the treatment of Hirschsprung’s disease (HD). Glial cell-derived neurotrophic factor (GDNF) and neurotrophin-3 (NT-3) are neurotrophic factors that play important roles in neuronal development, differentiation, survival and function. Meanwhile, GDNF mutations are a major cause of HD. In this study, BMSCs were transfected with eukaryotic expression plasmids co-expressing GDNF and NT-3, and the transfected cells displayed neuron-like changes after differentiation induced by fetal gut culture medium (FGCM). Immunofluorescence assay showed positive expression of the neuronal marker NSE and the enteric neuronal markers PGP9.5, VIP and nNOS. Reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of GDNF and NT-3 in transfected BMSCs. The present study indicates that genetically modified BMSCs co-expressing GDNF and NT-3 are able to differentiate into enteric neuronal cells and express enteric nerve markers when induced by FGCM. This study provides an experimental basis for gene therapy to treat enteric nervous system-related disorders, such as HD.

This study was aimed to explore the influence of breast cancer associated fibroblasts (CAFs) in migration and invasion of breast cancer cell line MCF-7, and investigate whether hepatocyte growth factor (HGF) is involved in this process. Primary breast CAFs and their corresponding normal breast fibroblasts (NFs) were obtained by collagenase digestion. On the basis of the co-culture, the migration and invasion capacity of MCF-7 cells was compared between CAFs and NFs by Transwell. The difference in the HGF expression between them was detected by ELISA. The secretion of HGF was knocked down by using RNA interference technology in CAFs. Then the changes of migration and invasion capacity of MCF-7 cells were investigated by Transwell. Eventually, we isolated high-purity CAFs and NFs, and the CAFs had a stronger ability in promoting MCF-7 migration and invasion than the NFs. ELISA results demonstrated that CAFs secreted higher HGF, and the capacity of MCF-7 migration and invasion was declined after knocking down the secretion of HGF in CAFs by RNA interference. It is suggested that CAFs can promote MCF-7 migration and invasion through HGF in vitro.

The prevalence of human papilloma virus (HPV)-16 in patients with cervical cancer, the physical status of HPV-16 in patients with cervical lesions, and the role of HPV-16 integration in cervical carcinogenesis were investigated. HPV genotyping was performed by using PCR approach with the primer GP5+/GP6+ and type-specific primer on biopsy specimens taken operatively from 198 women. Multiple PCR was done to detect physical status of HPV-16 in a series of cervical liquid-based cytology samples and biopsy specimens obtained from different cervical lesions with HPV-16 infection, including 112 specimens with cervical cancer, 151 specimens with CIN I, 246 specimens with CIN and 120 specimens with CINIII. The results showed that there were 112 cervical cancer samples (56.57% of total cervical cancer patients) with HPV-16 infection. The frequency of HPV-16 pure integration was 65.18% (73/112), 56.57% (47/120), 23.58% (58/246) and 7.95% (12/151) in cervical cancer, CINIII, CINII and CINI patients respectively. In situ hybridization was performed on some paraffin-embedded sections of CINII, CINIII and cervical cancer to verify the physical status of HPV-16 infection. Significant difference was observed between cervical cancer and CIN I, CINII, CINIII in the frequency of HPV-16 integration (P<0.01). It is suggested that HPV-16 is the most prevalent type and is associated with cervical cancer. In the case of HPV-16 infection there are close associations between the severity of cervical lesions and the frequency of HPV-16 integration. The application of testing HPV genotyping and physical status based on detection of HC-II HPV DNA would be in favor of predicting the prognosis of cervical precancerosis and enhancing the screening accuracy of cervical cancer.

In this study, real-time PCR and immunohistochemistry were used to detect coxsakie and adenovirus receptor (CAR) expression. Both localization and quantity were evaluated in the uteri obtained at days post coitus (dpc) 2.5, 4.5, 6.5, 8.5. Outcome of PCR was assessed by 2^−ΔΔCt method. Image Pro-Plus 6.0 software was used for quantifying mean density of CAR expression in immunohistochemical sections. We found relatively weak CAR expression in the mouse uteri during implantation window. PCR and immunohistochemistry revealed highest CAR expression was detected on dpc 2.5 followed by down-regulation of CAR at dpc 4.5 and 6.5 (with significant difference). At dpc 8.5, CAR expression was increased slightly again. It is concluded that during implantation, the expression of CAR mRNA and protein is declined, resulting in the impairment of tight junction between cavity epithelium cells. After implantation window closure, CAR appears again to maintain epithelium stability. CAR might play an important role during embryo implantation procedure.

In previous study, glutaric acid (GA) induced apoptosis of primary striatal neuron in vitro. In order to investigate the neurotoxic effects of GA on neonatal rat corpus striatum and the possible mechanism, 34 male pups were randomly assigned to NS group, low dose GA (LGA, 5 μmol GA/g body weight) group and high dose GA (HGA, 10 μmol GA/g body weight) group. These pups were subcutaneously administered with three injections from postnatal day 3 to 22 at 7:30 am, 15:00 pm and 22:30 pm and killed 12 h after the last injection. Microscopic pathology in corpus striatum was evaluated by HE staining. The apoptotic cells were identified by TUNEL staining. The transcript levels of caspase-3, 8, 9, Bax, Bcl-2 were detected by using real-time PCR and the protein levels of procaspase-3 and the active fraction were evaluated by Western blotting. In LGA and HGA groups, ventricle collapse, cortical atrophy by a macroscope and interstitial edema, vacuolations, widened perivascular space of bilateral striatum by a microscope were observed. TUNEL assay revealed that the apoptotic cells were increased in LGA and HGA groups. The transcript of caspase-3 was up-regulated to 2.5 fold, accompanied by the up-regulation of caspase-9, Bax and down-regulation of Bcl-2. The protein levels of procaspase-3 and the active fraction were up-regulated in LGA and HGA groups. The rat model for GA I showed mitochondrial apoptotic pathway may be involved in the GA-induced striatal lesion. Further studies should be taken to investigate the underlying mechanisms.

This study examined the expression pattern of programmed cell death 5 (PDCD5) in cochlear hair cells and spiral ganglion neurons (SGNs) and its association with age-related hearing loss in mice. Sixty C57BL/6J (C57) mice at different ages were divided into four groups (3, 6, 9 or 12 months). PDCD5 expression was detected by using immunohistochemistry, real-time PCR and Western blot. Morphological change of the cochleae was also evaluated by using immunoassay. The results showed that the expression of PDCD5 had a gradual increase with ageing in both protein and RNA levels in C57 mice, as well as gradually increased apoptosis of cochlear hair cells and SGNs. In addition, we also found that caspase-3 activity was enhanced and its expression was enhanced with ageing. It is implied that overexpression of PDCD5 causes the increase in caspase-3 activity and the subsequent increase of apoptosis in cochlear hair cells and SGNs, and thereby plays a role in the pathogenesis of presbycusis. Thus, PDCD5 may be a new target site for the treatment and prevention of age-related hearing loss.

The accuracy and repeatability of computer aided cervical vertebra landmarking (CACVL) were investigated in cephalogram. 120 adolescents (60 boys, 60 girls) aged from 9.1 to 17.2 years old were randomly selected. Twenty-seven landmarks from the second to fifth cervical vertebrae on the lateral cephalogram were identified. In this study, the system of CACVL was developed and used to identify and calculate the landmarks by fast marching method and parabolic curve fitting. The accuracy and repeatability in CACVL group were compared with those in two manual landmarking groups [orthodontic experts (OE) group and orthodontic novices (ON) group]. The results showed that, as for the accuracy, there was no significant difference between CACVL group and OE group no matter in x-axis or y-axis (P>0.05), but there was significant difference between CACVL group and ON group, as well as OE group and ON group in both axes (P<0.05). As for the repeatability, CACVL group was more reliable than OE group and ON group in both axes. It is concluded that CACVL has the same or higher accuracy, better repeatability and less workload than manual landmarking methods. It’s reliable for cervical parameters identification on the lateral cephalogram and cervical vertebral maturation prediction in orthodontic practice and research.

Previously, the choice of prosthetic implant-retained overdentures has depended on data from previous studies about the retention-fatigue strength of the attachment system selected. Little or no data have been available on the correlation between the attachment system selected and the overdenture support configuration. The purpose of the present study was to evaluate the retention force and fatigue resistance of three attachment systems and four support designs of overdenture prosthesis. Four lower edentulous acrylic models were prepared and eight combinations of attachments groups were investigated in the study. These included: O-Rings with mini-dental implants (MDIs), Dalbo elliptic with Dalbo Rotex and fabricated flexible acrylic attachments with both MDI and Dalbo Rotex. The study was divided into four test groups: groups A and B, controls, and groups C and D, experimental groups. Control group A contained three overdenture supports: two free standing MDIs in the canine region and at the midline, and one simulated tooth root with Dalbo Rotex screwed in. Control group B contained four overdenture support foundations: two free standing MDIs in the right canine region and the first premolar region, and two simulated tooth roots with Dalbo Rotex screwed in at the same MDI position, but on the left side of the model. Experimental group C contained three overdenture support foundations: two free standing MDIs in the canine region and at the midline, and one simulated tooth root with MDI screwed in. Experimental group D contained four overdenture support foundations: two free standing MDIs in the right canine region and the first premolar region, and two simulated tooth roots with MDIs screwed in at the same MDI position, but on the left side of the model. Each group was further divided into two subgroups according to attachment type used. Five samples were prepared for each group. Retention force (N) values were recorded initially (0 cycles) and after 360, 720, 1440 and 2880 insertion and removal cycles. During the tensile test a cross-head speed of 10 mm/min was applied. Values of absolute force (AF) and relative force (RF) were statistically analyzed by two-way ANOVA and multiple comparison Tukey’s tests between groups and cycles periods. The results of fatigue tests showed a 50% reduction in retention force in the subgroups with flexible attachments. A triangular design of overdenture support foundations with O-Ring attachments revealed the lowest value of AF and a relatively high reduction in RF. The four overdenture support designs with flexible acrylic attachments improved the retention force and reduced the fatigue retention. Furthermore, the results of the investigation demonstrate that flexible acrylic attachments for both teeth and implant-supported overdentures offer a wide range of retention forces.

As a nonparametric method, the Kruskal-Wallis test is widely used to compare three or more independent groups when an ordinal or interval level of data is available, especially when the assumptions of analysis of variance (ANOVA) are not met. If the Kruskal-Wallis statistic is statistically significant, Nemenyi test is an alternative method for further pairwise multiple comparisons to locate the source of significance. Unfortunately, most popular statistical packages do not integrate the Nemenyi test, which is not easy to be calculated by hand. We described the theory and applications of the Kruskal-Wallis and Nemenyi tests, and presented a flexible SAS macro to implement the two tests. The SAS macro was demonstrated by two examples from our cohort study in occupational epidemiology. It provides a useful tool for SAS users to test the differences among three or more independent groups using a nonparametric method.

In order to investigate the mortality of a cohort of chrysotile asbestos miners in China and evaluate its association with exposure to chrysotile, a fixed cohort of 1932 workers in chrysotile asbestos mine was established in 1981 and followed till June 1, 2010. Information on vital status, cause of death and smoking habits was collected. The workers were divided into two groups according to their exposure status. The exposed group was composed of frontline workers who worked directly on mining or processing asbestos products. The control group consisted of those who were not directly exposed to asbestos in their work. Standardized mortality ratio (SMR) was calculated according to Chinese national death rates. Cox proportional hazards model was applied to estimate the adjusted relative risks of deaths from major causes in exposed and control groups. The results of this study showed that main causes of mortality were malignant neoplasm, cardiovascular disease, cerebrovascular disease and respiratory disease for chrysotile miners. The mortality rate was 939.20 per 100 000 person-years for workers. The SMR for all causes of death was 1.46 in the cohort. Statistically significant mortality excesses were found for lung cancer (SMR=1.51), pulmonary heart disease (SMR=2.70), respiratory disease (SMR=1.93), asbestosis (SMR=9.62), and accident (SMR=1.59). The mortalities from malignant neoplasm, lung cancer, cerebrovascular disease and digestive disease in the exposed group were significantly higher than those in the control group. The findings indicate that chrysotile exposure is a risk factor for lung cancer, respiratory disease, cerebrovascular disease and digestive disease.

The deficiency theories of dyslexia are quite contradictory and the cross-cultural studies in recent years mainly focused on whether the dyslexics among cultures shared the same cognitive profile or just based on the language. This study used Near-Infrared Spectroscopy (NIRS) imaging to measure the regional cerebral blood volume (BV) and the changes of cerebral activation in the left prefrontal cortex of 12 Chinese dyslexic children and their 12 age-matched normal controls during the Paced Visual Serial Addition Test (PVSAT). Results showed that the scores of PVSAT of dyslexic children were significantly lower than those of the normal children (t=3.33, P<0.01). The activations of the left prefrontal cortex in the normal group were significantly greater than those of dyslexic children (all P<0.01). Our results indicated that Chinese dyslexia had a general deficiency in working memory and this may be caused by the abnormal metabolic activity of brain blood volume in the left prefrontal cortex and the deficits in brain function might be the basis of neuropathology of Chinese dyslexia. Present study supports the difference on brain activation of dyslexics from different languages may be caused by the same cognitive system related to reading.

Superior mesenteric vein (SMV) thrombosis is a relatively rare disease. Most patients may be successfully treated with anti-coagulation alone. However, bowel stricture may develop due to intestinal ischemia which may require surgical treatment. This report describes a rare case of small bowel stricture occurring one month after successful treatment of SMV thrombosis. After segmental resection of strictured bowel, the patient’s post-operative course was uneventful.

Postpartum inferior vena cava (IVC) thrombosis is a rare, but potentially life-threatening disorder. Here we reported one case of the youngest woman to date who presented with massive IVC thrombus extending from deep veins of the right leg to the level of the 11th thoracic vertebra, associated with asymptomatic pulmonary embolism.