Human interleukin-15 (hIL-15) is an important cytokine to activate endothelial cells and can be regulated by many other cytokines. The aim of this study is to examine the ability of interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) to induce the production of human interleukin-15 (hIL-15) and IL-15 receptor (IL-15Rα) by human umbilical vein endothelial cells (HUVECs). The data are summarized as follows: 1. Northern blot revealed that IL-15 mRNA was up-regulated by IFN-γ and TNF-α. 2. Intracellular IL-15 protein was visualized by fluorescence microscopy, whereas the expression of IL-15 on the surface of HUVECs was detected by fluorescence activated cell sorting (FACS), and no detectable IL-15 in the medium was verified by ELISA. 3. IL-15Rα was detected on the surface of HUVECs by FACS after IFN-γ and TNF-α stimulation, whereas Western blotting revealed that the elevated expression on surface IL-15Rα was not due to the increased protein expression. The conclusion demonstrated from our results is that IFN-γ and TNF-α play an important role in regulating the expression of IL-15 and IL-15Rα on the surface of HUVECs.

In order to identify novel genes involved in spermatogenesis, testis cDNA samples from Balb/C mice of different postnatal days were hybridized with the whole mouse genome Affymetrix chip to screen the testis-specific genes. The characteristics of the selected genes were analyzed by RT-PCR as well as other bioinformatic tools. A novel differentially expressed testis-specific gene (GenBank Accession No: NM_029042) in the developmental stages of testes was identified, and named TSCPA. Cellular mapping prediction of TSCPA indicated that its protein was probably expressed in nuclei, and one putative domain (aa 332–377) was anchoring domain of cAMP-dependent type II PK. The result of subcellular localization of GFP-TSCPA fusion protein in Cos-7 cells showed that TSCPA protein was expressed in nuclei. RT-PCR analysis revealed that TSCPA was expressed specifically in mouse and human testis. TSCPA gene was expressed weakly in 21-day-old mouse testis and the expression was increased gradually from 38th day to 6th month of mouse testes. No expression of hTSCPA was found in cryptorchidism and Sertoli-cell-only syndrome patients. It was concluded that the expression profile of TSCPA in human and mice indicated that TSCPA might play an important role in spermatogenesis.

Overexpression of human ether-à-go-go (eag) related gene (hERG) has been found in a broad range of human leukemia cell lines and primary human leukemia. The block of hERG protein might be a potential therapeutic strategy for leukemia. Gambogic acid (GA) has recently exhibited marked anti-tumor potency on solid tumors of various derivations. Here, we investigated the anti-leukemia effects of GA and its relation to the regulation of hERG in K562 leukemia cells in vitro. K562 cells were treated with various concentrations of GA (0.125–8.0 µmol/L) for 0–72 h. MTT assay was used to evaluate the inhibition effect of GA on the growth of K562 cells. Cell apoptosis was measured through both Annexin-V FITC/PI double-labeled cytometry and transmission electron microscopy. Cell cycle regulation was studied by a propidium iodide method. RT-PCR and Western blot were applied to detect the expression level of hERG in K562 cells. GA presented striking growth inhibition and apoptosis induction potency on K562 cells in vitro in a time- and dose-dependent manner. The IC₅₀ value of GA for 24 h was 2.637±0.208 µmol/L. Moreover, GA induced K562 cells arrested in G₀/G₁ phase, accordingly, cells in S phase decreased gradually, and no obvious changes were found in G₂/M phase cells. Under the transmission electron microscopy, apoptotic bodies containing nuclear fragments were found in GA-treated K562 cells. After treatment with GA of 2.0 µmol/L for 24 h, the percentage of apoptotic cells was increased from 4.09% to 18.47% (P<0.01). Overexpression of hERG channel was found in K562 cells, while GA could down-regulate it at both protein and mRNA levels (P<0.01). It was concluded that GA exhibited its anti-leukemia effects partially through down-regulating the expression level of hERG channel in K562 cells, suggesting that GA may be a potential agent against leukemia with a mechanism of blocking hERG channel.

The effects of two different histone deacetylase (HDAC) inhibitors, sodium butyrate (NaB) and trichostatin A (TSA),on apoptosis of human leukemic cells in vitro and the molecular mechanisms were investigated. The experiments were divided up 5 groups: control group, NaB group, TSA group, NaB+Z-VAD-FMK group and TSA+Z-VAD-FMK group. The apoptosis rate was determined by morphological analysis and flow cytomytry. The expression of Daxx, Bcl-2, and Bcl-xl proteins was detected by Western blot. NaB and TSA could induce the apoptosis of HL-60 and K562 cells, and Z-VAD-FMK caused a marked decrease in apoptosis induced by HDAC inhibitors. HDAC inhibitors could down-regulate the expression of Daxx protein, but had no significant influence on the expression of Bcl-2 and Bcl-xl proteins. The results suggested that NaB and TSA induce distinct caspase-dependent apoptosis of human leukemic cells through down-regulating the expression of Daxx protein in vitro.

The DNA damage, caused by cigarette smoking, can cause airway cell apoptosis and death, which may be associated with the development of chronic obstructive pulmonary disease (COPD). However, just 20%–30% smokers develop COPD, which suggests that different degrees of DNA repair cause different outcomes in smokers. X-ray repair cross-complementing group 1 (XRCC1), a base excision repair protein, has multiple roles in repairing ROS-mediated, basal DNA damage and single-strand DNA breaks. The present study investigated the association between polymorphism in XRCC1 (Arg399Gln) and susceptibility of COPD. A total of 201 COPD cases and 309 controls were recruited and frequency-matched on age and sex. XRCC1 genotype was determined by PCR-restriction fragment length polymorphism analysis. Overall, compared with those with the XRCC1 Arg/Arg genotype, the risk for COPD had no significant difference among individuals with Trp/Trp genotype. However, after stratifying by smoking status, in former smokers, compared with those with the XRCC1 Arg/Arg genotype, the risk for COPD was significantly reduced among individuals with Trp/Trp genotype (adjusted OR=0.22, 95% CI 0.06–0.85, P=0.028); after stratifying by smoking exposure, in light smokers, compared with those with the XRCC1 Arg/Arg genotype, the risk for COPD was significantly reduced among individuals with Arg/Trp genotype and Trp/Trp genotype (adjusted OR=0.39, 95% CI 0.16–0.94, P=0.036; 0.24, 95% CI 0.07–0.79, P=0.019, respectively). In conclusion, XRCC1 Arg194Trp genotype is associated with a reduced risk of developing COPD among former and light smokers.

The expression of angiopoietin-1 (Ang-1) and thrombospondin-1 (TSP-1) in 5/6 subtotal nephrectomy (STN) rats model, and its correlation to the renal microvasculature injury were investigated. Rat 5/6 STN model was established in adult male SD rats, and the sham-operated group and 5/6 STN group were set up. The renal function and histopathological changes were examined at the 1st, 2nd, 4th, 8th and 12th week after operation. The expression of Ang-1, TSP-1 and CD31 in renal tissues was detected by using immunohistochemistry. From 2nd to 8th week after operation, Ang-1 was significantly expressed in glomeruli of rats with STN. Ang-1 staining in glomeruli of STN group was increased significantly as compared with that in sham-operated group at 4th and 8th week after operation, and subsequently decreased after the 12th week. The expression of TSP-1 was increased significantly in STN group. As compared with sham-operated group, the CD31 expression was significantly down-regulated from the 2nd week. The expression of Ang-1 mRNA was detected by using RT-PCR at the same time points. The expression of Ang-1 mRNA in renal tissue of rats with STN was significantly up-regulated at the 2nd, 4th and 8th week after operation as compared with that in STN group at other time points or in sham-operated group at the same time points, while decreased evidently at the 12th week as compared with that in sham-operated group. It is concluded that there are changes in the mRNA expression of Ang-1, and the significant up-regulation of the expression of TSP-1 in renal tissue of rats with STN, which may be involved in the remnant renal microvasculature injury.

The effects of vascular endothelial growth factor (VEGF) on neural differentiation of human embryonic stem cells (hESCs) in vitro and the possible mechanism were observed. The hESCs lines, TJMU1 and TJMU2, were established and stored by our laboratory. hESCs differentiated into neuronal cells through embryonic body formation. In this induction process, hESCs were divided into three groups: group A, routine induction; group B, routine induction+10 ng/mL VEGF; group C, routine induction+10 ng/mL VEGF+10 ng/mL VEGFR2/Fc. OCT4, Nestin and GFAP in each group were detected by RT-PCR, and the cells expressing Nestin and GFAP were counted by immunofluorescence. The percentage of Nestin positive cells in group B was significantly higher than in groups A and C, while the percentage of GFAP positive cells in group B was significantly lower than in groups A and C (P<0.01). There was no significant difference between groups A and C (P>0.05). It was concluded that VEGF, via VEGFR2, stimulated the neural differentiation of hESCs in vitro.

Hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) are believed to play a major role in viral clearance and disease pathogenesis during HBV infection. To clarify the differences in host immune responses between self-limited and chronic HBV infections, we constructed three HLA-A*0201/HBV tetramers with immunodominant epitopes of core18–27, polymerase 575–583 and envelope 335–343, and analyzed the HBV-specific CTLs in peripheral blood mononuclear cells (PBMCs) from patients infected with HBV. The frequencies and expansion ability of HBV-specific CD8⁺ T cells in most self-limited HBV infected individuals were higher than those in chronic HBV-infected patients. HBV-specific CD8⁺ T cells could be induced by in vitro peptide stimulation from chronic patients with a low level of serum HBV-DNA but not from those with a high level of serum HBV-DNA. In chronic infection, no significant correlation was found either between the frequencies of HBV-specific CD8⁺ T cells and the viral load, or between the frequencies and the levels of alanine transaminase. Our results suggested that the frequencies of HBV-specific CTLs are not the main determinant of immune-mediated protection in chronic HBV infection and immunotherapeutic approaches should be aimed at not only boosting a HBV-specific CD8⁺ T response but also improving its function.

The pathogenesis of HBsAg (+)/HBsAb (+) double positive hepatitis B virus infection was investigated by simulating HBsAg/HBsAb coexistence in vitro and establishing HBsAg/HBsAb double positive model in vivo. Eukaryotic expression plasmids PCI-SY, PCI-adw, PCI-adr, PCI-ayw, which expressed S gene product of different serotypes, were constructed and transfected into HepG2 cells. Recombinant proteins were purified from the transfected cells. At the same time, HBsAg mouse antiserum was obtained by immunizing mice with PCI-SY plasmid. HBsAg/HBsAb coexistence was simulated using these antigens and antiserum. Furthermore, the expression plasmids expressing different serotypes of S gene product including PCI-adw, PCI-adr, and PCI-ayw were injected into mice via tail vein. HBsAg and HBsAb in mice sera were tested at the first and 7th day respectively after antigen plasmids injection. Both in vitro simulation and in vivo animal models demonstrated that HBsAg antigen and HBsAb of the same serotypes could not coexist, but HBsAg antigen and HBsAb of different serotype could coexist. HBsAg/HBsAb double positive hepatitis B virus infection could be due to infection of viruses of different serotypes.

The effects of highly active antiretroviral therapy (HAART) to patients with AIDS in Hubei province of China were investigated in order to provide scientific evidence to reinforce the management of HAART. Self-made questionnaires and descriptive method of epidemiology were used to collect and describe the changes of clinical symptoms, HIV RNA concentration, and immune function of patients with AIDS. After HAART, the effective rate of fever, cough, diarrhea, lymphadenectasis, weight loss, tetter, debility and fungous infection was 92.4%, 90.85%, 92.91%, 90.73%, 93.69%, 89.04%, 92.34%, and 83.1%, respectively. Of 117 patients with detected HIV RNA concentration, 41.03% had declined over 0.5 log, and 52.99% less than 0.5 log. CD4⁺T cell count was obviously increased: the average number after HAART for 3 or 6 months was 237/µL (26–755/µL) and 239/µL (17–833/µL), respectively. HAART can improve AIDS patients’ clinical symptoms, reduce HIV RNA concentration, and maintain immune function. It is very important for the effectiveness of HAART to raise clinical adherence of patients with AIDS and have a persistent surveillance.

To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis (2-D DIGE) and reversed phase high-performance liquid chromatography (RP-HPLC) followed by electrospray ionization mass spectrometry (ESI-MS) were used. Serum samples from renal allograft patients and normal volunteers were divided into three groups: acute rejection (AR), stable renal function (SRF) and normal volunteer (N). Serum samples were firstly processed using Multiple Affinity Removal Column to selectively remove the highest abundance proteins. Differentially expressed proteins were analyzed using 2-D DIGE. These differential protein spots were excised, digested by trypsin, and identified by RP-HPLC-ESI/MS. Twenty-two differentially expressed proteins were identified in serum from AR group. These proteins included complement C9 precursor, apolipoprotein A-IV precursor, vitamin D-binding protein precursor, beta-2-glycoprotein 1 precursor, etc. Vitamin D-binding protein, one of these proteins, was confirmed by ELISA in the independent set of serum samples. In conclusion, the differentially expressed proteins as serum biomarker candidates may provide the basis of acute rejection noninvasive diagnosis. Confirmed vitamin D-binding protein may be one of serum biomarkers of acute rejection. Furthermore, it may provide great insights into understanding the mechanisms and potential treatment strategy of acute rejection.

In order to improve the surface properties of PLGA polymer for a better material/cell interface to modulate the cells behaviors, we prepared a novel three-block copolymer, PLGA-[ASP-PEG], and immobilized an RGD-containing peptide, Gly-Arg-Gly-Asp-Ser-Pro-Cys (GRGDSPC) on the surface of it. Transforming growth factor-β1 (TGF-β1) was transfected into bone marrow stromal cells (MSCs) employed as seeded cells. Cell adhesion, spreading, proliferation and differentiation on this material were investigated. The results showed that the cell adhesive ratio on RGD-modified materials was higher than on un-modified materials (P<0.05). The extent of cell spreading was also wider on RGD-modified materials than on un-modified materials. Cell proliferation indices of transfected MSCs were increased as compared with the un-transfected MSCs (P<0.05). The ALP activities in the MSCs cultured with RGD-modified materials were higher than on un-modified materials after 14 days (P<0.05), and those in transfected MSCs were higher than in un-transfected MSCs (P<0.05). It was suggested that the combined use of RGD-modification and TGF-β gene transfection could improve the interaction of biomaterial and cells.

The aim of this study was to fabricate biomatrix/polymer hybrid scaffolds using an electrospinning technique. Then tissue engineered heart valves were engineered by seeding mesenchymal stromal cells (MSCs) onto the scaffolds. The effects of the hybrid scaffolds on the proliferation of seed cells, formation of extracellular matrix and mechanical properties of tissue engineered heart valves were investigated. MSCs were obtained from rats. Porcine aortic heart valves were decellularized, coated with poly(3-hydroxybutyrate-co-4-hydroxybutyrate) using an electrospinning technique, and reseeded and cultured over a time period of 14 days. In control group, the decellularized valve scaffolds were reseeded and cultured over an equivalent time period. Specimens of each group were examined histologically (hematoxylin-eosin [HE] staining, immunohistostaining, and scanning electron microscopy), biochemically (DNA and 4-hydroxyproline) and mechanically. The results showed that recellularization was comparable to the specimens of hybrid scaffolds and controls. The specimens of hybrid scaffolds and controls revealed comparable amounts of cell mass and 4-hydroxyproline (P>0.05). However, the specimens of hybrid scaffolds showed a significant increase in mechanical strength, compared to the controls (P<0.05). This study demonstrated the superiority of the hybrid scaffolds to increase the mechanical strength of tissue engineered heart valves. And compared to the decellularized valve scaffolds, the hybrid scaffolds showed similar effects on the proliferation of MSCs and formation of extracellular matrix. It was believed that the hybrid scaffolds could be used for the construction of tissue engineered heart valves.

To investigate the exon mutation of vitamin K-dependent gamma-glutamyl carboxylase (GGCX or VKDC) in patients with calcium oxalate urolithasis, renal cortex and peripheral blood samples were obtained from severe hydronephrosis patients (with or without calculi), and renal tumor patients undergoing nephrectomy. GGCX mutations in all 15 exons were examined in 44 patients with calcium oxalate urolithiasis (COU) by polymerase chain reaction (PCR) and denatured high pressure liquid chromatography (DHPLC), and confirmed by sequencing. Mutation was not found in all COU samples compared to the controls. These data demonstrated that functional GGCX mutations in all 15 exons do not occur in most COU patients. It was suggested that there may be no significant association between the low activity and mutation of GGCX in COU.

The correlation between the anatomic site of spinal cord injury and real-time conditions of bladder and urethral function was assessed in order to provide a reasonable basis for the clinical treatment of neurogenic bladder. A total of 134 patients with spinal cord injuries (105 males, 29 females; averaged 34.1 years old) were involved in this retrospective analysis, including urodynamic evaluation, clinical examination and imaging for anatomical position, and Bors-Comarr classification. The associations between the levels of injury and urodynamic findings were analyzed. The results showed that mean follow-up duration was 16.7 months (range 8–27 months). Complete spinal cord injuries occurred in 21 cases, and incomplete spinal cord injuries in 113 cases. Of the 43 patients with upper motor neuron (UMN) injuries, hyperreflexia and (or) detrusor sphincter dyssynergia were demonstrated in 30 (69.8%), 31 (72.1%) suffered low bladder compliance (less than 12.5 mL/cmH₂O), 28 (65.1%) had high detrusor leak point pressures (greater than 40 cmH₂O), and 34 (79.1%) had residual urine. Of the 91 patients with lower motor neuron (LMN) injuries, areflexia occurred in 78 (85.7%), high compliance in 75 (82.4%), low leak point pressures in 80 (87.9%), and residual urine in 87 (95.6%), respectively. The associations between the anatomical site of spinal cord injury and urodynamic findings were ill defined. In patients with spinal cord injury, this study revealed a significant association between the level of injury and the type of voiding dysfunction. The anatomical site of spinal cord injury can not be predicted in real-time condition of bladder and urethral function. Management of neurogenic bladder in patients with spinal cord injury must be based on urodynamic findings rather than inferences from the neurologic evaluation.

The action mechanism of matrix metalloproteinases-2 (MMP-2) and tissue inhibitor of metalloproteinases-2 (TIMP-2) in the genesis, development and degeneration of haemangioma was investigated by detecting their expression in the tissue of haemangioma in different phases by using the immunohistochemistry. Fifty paraffin-embedded specimens of skin capillary haemangioma were collected, which were documented in the Department of Pathology, Renmin Hospital of Wuhan University from 2000 to 2006. All samples were stained by regular HE method, and proliferative cell nuclear antigen (PCNA) was tested by immunohistochemical S-P method. The samples were classified according to the Mulliken criteria and the expression pattern of PCNA. Immunohistochemical S-P method was applied to detect the expression of MMP-2 and TIMP-2 in proliferative and degenerative phases of cutaneous capillary haemangioma, and in normal skin tissues. In combination with the detection of the expression of factor VIII-related antigen, it was verified that in haemangioma tissues, the cells expressing MMP-2 and TIMP-2 were vascular endothelial cells. The MMP-2 and TIMP-2 expression was quantitatively analyzed by image analysis system (HPIAS-1000), and one-way ANOVA(107) and SNK(q) test were done to analyze average absorbance (A) and positive area rate of immunohistochemically positive particles by using SPSS11.5. The results showed: (1) Among 50 samples of haemangioma, there were 26 proliferative haemangiomas, and 24 degenerative haemangiomas, respectively; (2) The expression of MMP-2 was weak in normal vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression of MMP-2 in proliferative group was significantly higher than in degenerative group and control group (normal skin) (P<0.05), but there was no statistically significant difference between the latter two groups; (3) TIMP-2 was highly expressed in normal tissues, degenerative vascular endothelial cells, cytoplasm of connective tissues and extracellular matrix around blood vessels. The expression level of TIMP-2 in proliferative phase was significantly lower than in degenerative phase (P<0.05), and the expression of TIMP-2 in proliferative phase was significantly different from that in degenerative phase and normal tissues (P<0.05). It was concluded that in proliferative phase of haemangioma, MMP-2 may promote over-proliferation of endothelial cells of haemangioma, and in degenerative phase, TIMP-2 can inhibit the proliferation of endothelial cells of haemangioma. The two substances play important roles in the genesis, development and degeneration of haemangiomas.

In order to investigate the effects of vector-based hairpin small interference RNA (shRNA) on the reversal of multi-drug resistance (mdr) of A2780/Taxol cells, a novel vector pEGFP-H1/mdr1 containing mdr1-shRNA targeting at position 2943–2963 of mdr1 was designed and synthesized. Subsequently, A2780/Taxol cells were transfected with pEGFP-H1/mdr1, and the expression of mdr1 mRNA and P-gp was detected by using RT-PCR and Western blot respectively. MTT was used to measure the 50% inhibition concentration (IC₅₀) of Taxol to A2780/Taxol cells. The results showed that at the 24th and 48th h after transfection, the expression of mdr1 mRNA was decreased to (52.1±1.0)% and (0.01±1.7)%, and that of P-gp decreased to (88.3±2.1)% and 0%, respectively. At the 48th h after transfection, the relative reversal rate of A2780/Taxol cells to Taxol was 69.54%. In vivo, the nude mice xenografts were injected with pEGFP-H1/mdr1, and then administrated Taxol. The tumor volume in pEGFP-H1/mdr1-transfected group was significantly reduced as compared with that in blank control group or pEGFP-H1-transfected group (807.20±103.16 vs 1563.78±210.54 or 1480.78±241.24 mm³, both P<0.01). These results suggested that transfection of pEGFP-H1/mdr1 could efficiently down-regulate the expression of mdr1 mRNA and P-gp in A2780/Taxol cells, and effectively restore the sensitivity of A2780/Taxol cells to Taxol both in vitro and in vivo.

The recombinant plasmids pGenesil-1-BIRC71 and pGenesil-1-BIRC72 were transfected into Hela cells and cisplatin was added with different concentrations in order to study the inhibitory effects of Livin gene, increase the apoptosis induced by cisplatin, and detect the expression of Bcl-2, Bax, caspase-3, and survivin genes. The pGenesil-1-BIRC71 and pGenesil-1-BIRC72 were transfected into Hela cells, and the expression levels of Livin, Bcl-2, Bax, caspase-3, and survivin genes were detected by using fluorescence quantitative real-time PCR. Then cisplatin at different concentrations (3.0, 6.0 and 9.9 µg/mL) was added into the transfected Hela cells, and 24, and 48 h later, the apoptosis rate was measured by flow cytometry. After transfection of pGenesil-1-BIRC71 and pGenesil-1-BIRC72 into Hela cells, the expression level of Livin gene was obviously reduced, and the apoptosis rate was significantly increased in transfection group as compared with control group (P<0.05). Cisplatin could increase the apoptosis rate in a dose- and time-dependent manner. After cisplatin was added, the expression levels of Bcl-2 mRNA were reduced, and those of Bax, caspase-3, and survivin mRNA were increased in transfection group as compared with those in control group (P<0.05). It was concluded that shRNA expression vector targeting Livin gene could inhibit the expression of Livin gene in Hela cells and enhance the apoptosis induced by cisplatin, which was related to the decreased expression of Bcl-2 and activation of Bax and caspase-3. Survivin might play an important role as an antagonist in the process of apoptosis induction.

The effects of leptin on cytotrophoblast proliferation and invasion activity were investigated. Immunohistochemistry was used to determine the placental expression of leptin in first-trimester pregnancy. By using RT-PCR and quantitative real-time PCR, the expression of leptin in cytotrophoblast and the effect of leptin on cytotrophoblast secretion were detected. The potential of cell proliferation, invasiveness and migration was assessed by MTT, Transwell invasion assay and migration assay respectively when the cytotrophoblast was cultured with different concentrations of leptin. The results showed that: (1) Leptin was distributed diffusely around cell membrane, in cytoplasma, and on nuclear membrane of cytotrophoblast; (2) Leptin mRNA was expressed in cytotrophoblast. Ten ng/mL leptin could promote the secretion of cytotrophoblast significantly (P<0.01); (3) After culture with different concentrations of leptin for 24 h or longer, the proliferation of cytotrophoblast was inhibited, while in 24 h leptin could promote cytotrophoblast invasion and migration. Leptin at a concentration of 500 ng/mL could promote cytotrophoblast invasiveness and migration significantly as compared with controls (P<0.05). It was suggested that leptin could inhibit cytotrophoblast proliferation, and promote cytotrophoblast invasion and migration activity.

There has been emergence of evidence suggesting that specific variants of the vascular endothelia growth factor (VEGF) family, based on their ability to regulate angiogenesis, would be pivotal in the pathogenesis of endometriosis. This study was aimed at determining whether high levels of VEGF-A could be found in the serum and peritoneal fluid (PF) of patients with endometriosis. VEGF-A levels were measured by enzyme-linked immunosorbent assay (ELISA) in serum and PF from 46 patients with surgically confirmed endometriosis, and 40 controls with no clinical evidence of the disease or detectable endometriotic lesions at the time of surgical examination. The results showed the mean VEGF-A levels were significantly higher in the serum and PF of patients with endometriosis than in the controls. The VEGF-A levels in the serum and PF of patients with severe endometriosis (stages III–IV) were significantly higher than in those with minimal endometriosis (P<0.001). It was concluded that endometriosis was associated with significant modulation in the levels of circulating VEGF-A.

The relationship between the expression of resistin in polycystic ovary syndrome (PCOS) and insulin resistance was investigated. The plasma resistin concentrations in 35 patients with PCOS and 40 controls were measured by ELISA. Luteinizing hormone (LH), follicle-stimulating hormone (FSH), and fasting insulin (FIN) were tested by radioimmunoassay. Insulin resistance index (HOMA-IR) was calculated. Fasting plasma glucose (FPG) was determined by oxidase test. Western blot and reverse transcriptase PCR (RT-PCR) methods were used to detect the expression of resistin in adipose tissues. The levels of plasma resistin, LH, LH/FSH and FIN and HOMA-IR in patients with PCOS were significantly higher than those in control group (all P<0.05). Plasma resistin was correlated positively with FPG, FIN, HOMA-IR, LH and LH/FSH (r=0.56, 0.60, 0.65, 0.48, and 0.42 respectively). Resistin protein and mRNA expression levels in patients with PCOS were significantly higher than those in normal tissues (all P<0.01). It was concluded that resistin might be involved in the pathogenesis of insulin resistance of PCOS.

The relationship between Ala/Ser polymorphism in 133 codon of exon 3 region of the RASSF1 gene and genetic susceptibility of lung cancer in Hubei province Han population was investigated by a case-control study. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was adopted to analyze the polymorphism of codon 133 of exon 3 in the RASSF1 gene of 100 pathologically diagnosed lung cancer patients, and 100 healthy controls. The relationship between different genotypes and the susceptibility of lung cancer was analyzed. Among 200 blood samples from Han people in Hubei Province, including 100 from lung cancer patients and 100 from healthy controls, the frequencies of Ala/Ala, Ala/Ser, Ser/Ser genotype of the RASSF1 in lung cancer patients were 83%, 16%, 1%, and those in healthy controls was 93%, 7%, 0% respectively, with the difference being statistically significant between two groups (P<0.05). The individuals with Ala/Ser genotype had higher risk of suffering from lung cancer, with an OR of 2.341, and 95% CI of 1.009–6.393 respectively. It was concluded that RASSF1Ala133Ser was a susceptible genetic factor of lung cancer. Ala/Ser genotype increased the risk of lung cancer.

The effects of Wumeiwan (WMW) on TNF-α, IL-6, IL-8, IL-10 and NF-κBp65 in rats with ulcerative colitis (UC) were investigated, the curative effectiveness of WMW vs salicylazosulfapyridine (SASP) was compared, and the action mechanism was analyzed. Fifty-Six Sprague-Dawley (SD) rats were randomly divided into four groups (n=14 in each group, with equal ratio of male and female): normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene (DNCB) immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW respectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-α, IL-10 and the expression of NF-κBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-α were significantly increased (P<0.01), while those of IL-10 significantly reduced (P<0.01) after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-α were obviously lower, but the level of IL-10 was obviously higher in WMW and SASP groups than those in model group (P<0.05). The levels of IL-6, IL-8, and TNF-α were lower, while the level of IL-10 was higher in WMW group than in SASP group. NF-κBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-κBp65 in WMW group and SASP group was obviously lower than in model group (P<0.01), and there was significant difference between WMW group and SASP group (P<0.05). It was concluded that rat UC model was established successfully. WMW could up-regulate the expression of IL-10, down-regulate the expression of TNF-α, IL-6, IL-8, and inhibit the NF-κBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.

To re-identify and further group 25 isolates of Trichosporon spp. identified morphologically previously, sequences of D1/D2 region of large subunit (LSU) of ribosomal DNA (rDNA) of 25 tested strains for identification and those of ribosomal intergenic space 1 (IGS1) region of 11 strains for subgrouping were detected. The identifications of tested strains were changed except 6 strains. According to the alignment of the IGS1 region, 6 T. asahii isolates tested fell into 4 groups and 5 T. faecale isolates into 3 groups. Polymorphism of 2 T. japonicum isolates was found in 10 positions. With the alignments obtained in this research compared with the relative GenBank entries, it was found that T. asahii, T. faecale and T. japonicum species were divided into 7, 3 and 2 subtypes respectively. Morphological and biophysical methods are not sufficient for Trichosporon spp. identification. Sequencing becomes necessary for Trichosporon diagnosis. There is obvious diversity within a species.

This study evaluated the efficacy of rabdosia rubescens against gingivitis and compared the therapeutic efficacy of different dosage forms of rabdosia rubescens. A multi-center, randomized, double-blind, double-simulation, positive-controlled and parallel trial was conducted. A total of 136 patients exhibiting clinical symptoms of gingivitis were enrolled. The subjects were randomly assigned to two groups: test group (n=67), in which rabdosia rubescens drop pill (960 mg) and 4 tablets of simulation agent of rabdosia rubescen were orally given to the subjects three times a day for 5 days; and control group (n=69), in which the subjects were administered the tablets of rabdosia rubescens (1000 mg) and 24 drop pills of simulation agent of rabdosia rubescens thrice daily for 5 days. The experimental protocols and diagnostic criteria were established by expert panel prior to the experiment. The clinical symptoms were graded according to the severity of the disease and quantified. The total scores and scores for each clinical symptom of gingivitis were assessed at baseline and on the 6th day post-treatment. The therapeutic efficacy was compared between the two groups and in each group itself before and after the treatment. The results showed that in the two groups, the subjects who were given rabdosia rubescens, drop pill or tablet, had a decrease in total scores and scores for each clinical symptom when compared with those before treatment (P<0.01). There was significant difference in the therapeutic efficacy between the test group and the control group with the efficacy rate being 92.54% and 79.71% respectively (P<0.05). It was concluded that rabdosia rubescens showed great promise in treating gingivitis. And rabdosia rubescens drop pill was more efficacious than rabdosia rubescens tablet.

The clinically applied value of myocardial perfusion and systolic function in patients with coronary artery disease after coronary artery bypass surgery using real-time myocardial contrast echocardiography (RT-MCE) combined with two-dimensional strain echocardiography was assessed. Twenty patients underwent intravenous RT-MCE by intravenous injections of SonoVue before and after coronary artery bypass surgery. Two-dimensional images were recorded from the left ventricular four-chamber view, two-chamber view and the apical view before, and two weeks and three months after coronary artery bypass surgery, and the peak systolic longitudinal strain was measured. The results showed that myocardial perfusion was significantly increased after coronary artery bypass surgery in about 71.6% segments. In the group that myocardial perfusion was improved, the peak systolic longitudinal strain three months after bypass surgery was significantly higher than that before operation [(−15.78±5.91)% vs (−10.45±8.31)%, P<0.05]. However, the parameters did not change in the group without myocardial perfusion improvement [(−10.33±6.53)% vs (−9.41±6.09)%, P>0.05]. It was concluded that whether or not the improvement of myocardial perfusion can mirror the recovery trend of regional systolic function, two-dimensional strain echocardiography can observe dynamic change of regional systolic function. The combination of myocardial perfusion with two-dimensional strain echocardiography can more accurately assess the curative effectiveness of coronary artery bypass surgery.

The left ventricular radial strain in the inner and outer layers was evaluated by using two-dimensional speckle tracking imaging (2DS). Twenty-five piglets were studied. The short axis views were acquired. Peak systolic radial strain was measured from 6 circumferential points related to 6 standard segments in the inner and outer layers respectively using 2DS methods. The peak positive first derivative (dp/dt) of left ventricular pressure was compared to the radial strain from 2DS. The inner band showed higher peak radial strain values as compared to the outer band at all of the segments (P<0.0001), but the differences had significance just in anteroseptal, posterior, inferior and septal segments (P<0.05). Good correlation could be found between radial strain of inner and outer layers and peak dp/dt (P<0.001). These preliminary results showed that the degree of local deformation or wall thickening of the ventricular wall in its inner layer was more obvious than its outer layer. It is suggested that the 2DS technique is useful and sensitive for better understanding the regional and global myocardial motion and its relationship to the complex architecture of myocardium.

In order to explore a dose distribution verification procedure of intensity modulated radiation therapy (IMRT) for nasopharyngeal carcinoma (NPC) and establish its evaluation criteria, we performed 35 two-dimensional (2D) patient-specific IMRT verifications over the year 2006. The percent of pixels passing γ and the normalized agreement test (NAT) index were mainly used to represent the agreement between the measured and computed dose distributions with three criteria (2%/2 mm, 3%/3 mm and 5%/3 mm) as recommended in the literature. The results were that all cases passed through verifications with three criteria except that the NAT index of one case was beyond the limitation, and the three tolerance levels of 2%/2 mm, 3%/3 mm and 5%/3 mm produced similar clinical verification results but led to different percent of pixels passing γ and NAT index. Our data showed that the percent of pixels passing γ and the NAT index were complementary to evaluate future IMRT verifications as two significant metrics. Due to the influence of the noise and the trait of the software, we considered an IMRT plan as acceptable in case of the percent of pixels passing γ >95% and the NAT index <5 with the 5%/3 mm criteria for IMRT patient-specific quality assurance (QA).

Paget’s disease of the breast is an uncommon disorder that accounts for 1% to 3% of all mammary tumors. The incidence of underlying carcinoma associated with Paget’s disease has been reported in 82% to 100% of cases. The finding of underlying carcinoma reaches almost 100% when a palpable lump is also present. In this rare case, we described a patient presenting with Paget’s disease but no palpable lump. However, we found 11 independent regions which were all invasive ductal carcinoma after the operation. Considering this patient, we should pay more attention to a multifocal and multicentric breast carcinoma associated with Paget’s disease. Furthermore, we believe the mammography examination and a modified radical mastectomy are the most appropriate treatments for this population in clinical practice.