Objective To investigate the effects of bisphenol A on SD rat Sertoli cell function and the injury mechanism. Methods SD rat Sertoli cells were treated with bisphenol A at different doses,and the control group,solvent control group were set. Sertoli cell proliferation,cell cycle and PCNA,Vimentin protein expression. were observed. Results The survival rate of Sertoli cells in the treated groups was significantly lower than the control group (P

Objective To study the effects of siltuximab on the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (Stat3) signaling pathway in ovarian epithelial carcinoma.Methods (1) Expressions of IL-6 in ovarian cancer patient specimens were assessed by immunohistochemistry.(2) Expression of phosphorylation Stat3 (pStat3) protein in siltuximab and IL-6 treated SKOV3 cell lines was determined by western blot, and expression levels of Stat3-induced bcl-XL,MCL-1, survivin, in siltuximab treated SKOV3/TR and CAOV3/TR cells lines were also determined by western blot.(3) Real-time image analysis was used to study the nuclear translocation of pEGFP-Stat3 fusion protein in ovarian cancer cell line SKOV3-pEGFP-Stat3 treated with siltuximab and IL-6.(4)Paclitaxel sensitivity in siltuximab treated SKOV3/TR and CAOV3/TR cell lines were assessed using the methyl thiazolyl tetrazolium (MTT).The 50% inhibiting concentration ( IC50 ) was defined as the paclitaxel concentration required to decrease the A490 value to 50%.Results ( 1 ) There were significantly difference in IL-6 staining density and the positive rate of IL-6 protein stained among the metastatic, and drug-resistant recurrent tumors,and matched primary tumors [69%(18/26)] vs.77% (20/26)vs.23% (6/26), P＜0.05].(2)A clear increase in Stat3 phosphorylation levels was observed in the IL-6-treated SKOV3 cell lines as compared to the SKOV3 cell lines.When the IL-6-treated SKOV3 cells were incubated with siltuximab with a range of concentrations of 0.001,0.01,0.1, 1.0 and 10 μg/ml, there were trends toward reduced pStat3 expression in the treated cell lines.Compared without treatment with siltuximab, the expression of the anti-apoptotic proteins MCL-1, bcl-XL and survivin in SKOV3/TR and CAOV3/TR cell lines were significantly decreased after treated with siltuximab.(3) In resting cells, the majority of pEGFPStat3 was cytoplasmic until the addition of human IL-6, which promptly induced the translocation of fluorescent Stat3 molecules to the nucleus.Exposure of cells to siltuximab with a range of concentrations of 0.001, 0.01,0.1, 1.0 and 10 μg/ml, followed by an incubation in IL-6 significantly reduced pEGFP-Stat3 nucleocytoplasmic translocation.(4) MTT cytotoxicity assay demonstrated that siltuximab increased paclitaxel-induced cell death and partially overcame paclitaxel resistance.Treated with siltuximab ( 1 and 10 μg/ml) ,the paclitaxel IC50 value of siltuximab in SKOV3/TR (0.49, 0.19 μg/ml) and CAOV3/TR (0.0010, 0.0008 μg/ml) cells were significantly lower than those in untreated cells (0.71,0.0021 μg/ml;all P ＜ 0.05).Conclusions These results demonstrated that siltuximab effectively block the IL-6 signaling pathways, which .Blockage of IL-6 signaling may provide benefits for the treatment of ovarian cancer.

Objective To investigate the effect of MMP-9 on severe acute pancreatitis(SAP) associated with lung injury,and the preventive function of MMP-9 inhibitor (BB-94) in SAP associated with lung injury in rats. Methods SD rats were randomly divided into three groups: Shamed-operated groups (control groups,n=10);SAP groups (n=14);SAP with MMP-9 inhibitor preconditioning groups (BB-94 group,n=15). The samples for test of the wet weigh index of lung,serum amylase level,white cell count,the number of cells and protein in bronchoalveolus lavage fluid,histologic scoring of lung injury and MMP-9 in lung were detected 24 hours after SAP models were set up.Results The parameters mentioned above in SAP group were significant higher than those in controll group and BB-94 group (P

Objective To investigate the effect of diabetes on short-term prognosis of transient ischemic attack (TIA) in elderly patients.Methods From January 2006 to June 2010,126 patients with TIA aged over 60 years were selected.Patients were divided into diabetic group and non-diabetic group according to past history,blood glucose and glycosylated hemoglobin levels.The cumulative ischemic stroke incidences were analyzed by Kaplan-Meier survival analysis 30 days and 90 days after the first TIA.The risk factors for short-term stroke after TIA were analyzed by Cox regression analysis.Results Among 126 patients with TIA,31 cases (24.6％) had diabetes.The cumulative ischemic stroke incidences were significantly higher in diabetic group than in non-diabetic group 30 days and 90 days after the first TIA (54.8％ vs.22.1％,61.3％ vs.28.4％,both P＜0.01).Cox regression analysis revealed that diabetes and cerebral arterial stenosis were the risk factors for recurrent stroke within 90 days.Conclusions The short-term stroke incidence is significantly higher in elderly diabetic patients than in non-diabetic patients.Diabetes is the independent risk factor for recurrent stroke after TIA.

Objective To explore the effect of pain intervention on limb function exercises in patients with peripheric fractures of knee joint.Methods 60 patients with peripheric fractures of knee joint were randomized in equal number into the observation group and control group.Both groups took functional exercises for affected limbs.Besides,the former and latter groups were administered with celecoxib at a dosage of 200mg twice a day from pre-operation to discharge and after operation,respectively.The two groups were compared in terms of pain degree at different time points as well as the functional recovery of affected limbs.Results The observation group was lower in pain scores than the control group at hours 6,8,12,24,36 and 48.The active and passive motions of the affected limbs in the observation group were significantly better than those in the control at days 1,2,3,4 and 5(all P<0.001).Conclusion Pain intervention with celecoxib before operation may help patients to take functional exercises as early as possible,promoting the rehabilitation of functions of affected limbs.

This study was aimed to establish an identification method for Spatholobus suberectus and its adulterants by near-infrared spectroscopy (NIRS). Near-infrared diffuse reflection spectroscopy (NIRDRS) spectra of different S. suberectus and its adulterants were acquired by using OPUS INDENT analysis software. NIRDRS spectra clustering analysis model and identification model were established and verified. The results showed that S. suberectus from dif-ferent regions and its adulterants were identified successfully by clustering analysis model and identification model. It was concluded that Spatholobus suberectus and its adulterants can be identified rapidly and non-destructively by NIRS.

OBJECTIVETo compare the changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segment.

METHODSTwenty-one New Zealand white rabbits which were randomly divided into organ group with 8 rabbits and segment group with 13 rabbits. Fifty intervertebral discs and 50 spinal motion segments were harvested respectively under aseptic conditions from two groups. These specimens were maintained in organ culture with hyperosmotic media (410 mOsm/kg), then 10 discs of the two groups were observed respectively by HE staining, immunohistochemistry of collagen type III, proteoglycan content and cells viability of nucleus pulposus before culture and at 3, 7, 14, 21 days after culture.

RESULTSHE staining showed the intervertebral disc tissue structure remained intact after culture of 21 days organ group and 14 days segment group,but there was severely degenerated of 21 days segment group. The intensity value of type II collagen immunohistochemical staining in the nucleus pulposus were not changed significantly between 21 days organ group and 14 days segment group (P > 0.05), but the staining of segment group at 21 days became shallower, there was significant difference compared with before each time points and organ group at 21 days (P < 0.05). PAS/AB staining of proteoglycan of nucleus pulposus showed that there were not decrease of tinting strength of two groups within 7 days, but the strength weakened slightly of two groups at 14 days, and the tinting strength became weaker at 21 days segment group, the change is more obvious than the organ group. The intensity value of fluorescence staining of nucleus pulposus cells was not changed significantly within 7 days of two groups (P > 0.05), the intensity value decreased slightly at 21 days organ group and 14 days segment group, but there were no significant difference compared with before time points (P > 0.05) however at 21 days segment group the intensity decreased as cells viability of nucleus pulposus decreased,and there was a significant difference compared with before each time points and organ group at 21 days (P < 0.05).

CONCLUSIONIt is not obviously degenerated of the discs of organ group cultured within 21 days and segment group cultured within 14 days, but there was significant degeneration of the intervertebral disc of segment group after cultured 21 days, so the rabbit spinal motion segment can be used on research about the biomechanics of intervertebral disc as a vitro experimental model within 14 days.

Animals ; Collagen Type II ; analysis ; Female ; Immunohistochemistry ; Intervertebral Disc ; chemistry ; pathology ; Male ; Organ Culture Techniques ; Rabbits

Objective To investigate the expression of microRNA-106b(miR-106b)in the placentas of patients with pre-eclampsia and its relationship with matrix metallopeptidase(MMP)-2,and its effect on the invasion and proliferation of trophoblasts. Methods (1) Placental tissues were collected from patients with mild pre-eclampsia (mPE, n=30) , severe pre-eclampsia (sPE, n=30) and normal pregnant women (n=40). Human choriocarcinoma cell lines JAR and JEG3 were assigned to the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group, respectively. (2) The target gene of miR-106b(such as MMP-2) was predicted by bioinformatics. Dual-luciferase reporting system was used to verify the regulation of miR-106b on the expression of MMP-2. (3) The expressions of miR-106b and MMP-2 were measured by quantitative real-time PCR (qRT-PCR) and western blot. (4) Cell proliferation was determined by MTT assay. (5) Invasive activities in each group were assessed by cell transwell invasion assays. Results (1) Predicting result of bioinformatics indicated that MMP-2 was one of the target genes of miR-106b. Dual-luciferase activity assay demonstrated that MMP-2 was the direct target of miR-106b(P＜0.01).(2) The results of qRT-PCR.①The expression of miR-106b in the placentas of mPE, sPE, normal pregnant women were 2.89±0.04, 1.96±0.03, 1.01±0.03, respectively (P＜0.05). And the expression of MMP-2 mRNA in the placentas of mPE, sPE, normal pregnant women were 1.87±0.05, 0.69±0.03, 2.78±0.03, respectively (P＜0.05).②The expression of miR-106b in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 2.39 ± 0.03, 1.03 ± 0.04, 0.73 ± 0.03, 1.11 ± 0.04, respectively (P＜0.05). And its expression in the JEG3 cell line were 2.17±0.04, 1.18±0.04, 0.61±0.03 and 1.22±0.03, respectively (P＜0.05). ③The expression of MMP-2 mRNA in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 0.45±0.15, 1.02±0.03, 2.28±0.03, 1.11±0.03, respectively (P＜0.05). And its expression in the JEG3 cell line were 0.58±0.03, 1.25±0.15, 2.25±0.03, 1.21±0.03, respectively (P＜0.05). (3) The results of western blot.①The expression of MMP-2 protein in the placentas of mPE, sPE, normal pregnant women were 1.63 ± 0.04, 0.55±0.03, 2.82±0.03, respectively (P＜0.05).②The expression of MMP-2 protein in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 0.41 ± 0.03, 0.97 ± 0.03, 2.25 ± 0.03, 1.01 ± 0.03, respectively (P＜0.05). And its expression in the JEG3 cell line were 0.53±0.03, 1.20±0.03, 2.31±0.04, 1.19±0.03, respectively (P＜0.05). (4) miR-106b could inhibit the proliferation of JAR and JEG3 cells, cell proliferation rates in the miR-106b mimics group were lower than that in the mimics negative control group (P＜0.05). And cell proliferation rate in the miR-106b inhibitor group was higher than the inhibitor negative control group (P＜0.05) . (5) The numbers of JAR cell that passed the membrane in the miR-106b mimics group, the mimics negative control group. The miR-106b inhibitor group and the inhibitor negative control group were 61±15, 79±13, 134±13, 80±12, respectively( P＜0.05). And the numbers of JEG3 cell that passed were 57±12, 71±15, 128±15, 70± 14, respectively (P＜0.05). Conclusion The miR-106b could inhibit the invasion and proliferation of JAR and JEG3 cells through targeting MMP-2, and have a relationship with the pathogenesis of pre-eclampsia.

Objective To evaluate the quality of Radix et Caulis Ilicis Asprellae from Pingyuan planting base and Chinese herbal medicine market. Methods The water- and alcohol-soluble extracts from 19 batches of Radix et Caulis Ilicis Asprellae medicinal materials were detected according to Appendix ⅨH, ⅩA of the Chinese Pharmacopoeia ( 2010 edition). And the quality of the medicinal materials was evaluated by microscopic identification technology according to the method for Radix et Caulis Ilicis Asprellae recorded in Guangdong Provincial Chinese Medicine Standard, and then thin layer chromatography ( TLC) was optimized to establish the high performance liquid chromatography (HPLC) fingerprint. The HPLC was performed on Waters XBridgeTM C18 column (250 mm × 4.6 mm, 5μm) with acetonitrile(A)-0.2% (v/v) phosphorus acid (B) as the mobile phase by gradient elution, flow rate was 1.0 mL/min, and detection wavelength was 220 nm. Results The results of sample characters, TLC and microscopic identification showed that the samples of Radix et Caulis Ilicis Asprellae in Chinese herbal medicine markets were certified products, but stems and roots were blended. Seven common peaks were showed by HPLC and confirmed by similarity analytical software. The similarity of 15 batches of planting base samples was all above 0.9. Of 19 batches of the commercial samples, the similarity of 11 batches was above 0.9. The alcohol-soluble extract contents were in the range of 64.55 mg/g to 186.18 mg/g. Conclusion The medicinal materials of Radix et Caulis Ilicis Asprellae from Chinese herbal medicine market are certified products, but the qualities vary greatly for the blending of stems and roots and inadequate growth years. The quality of materials from planting base is better. The established method is helpful for the quality evaluation and control of Radix et Caulis Ilicis Asprellae.

Objective To establish the fingerprints and formononetin content determination method for Caulis Spatholobi from different habitats by high performance liquid chromatography ( HPLC) , thus to control the quality of Caulis Spatholobi. Methods Reversed phase-high performance liquid chromatography (RP-HPLC) for fingerprint was performed on Feini Gen RedClassical AQ-C18 column ( 4.6 mm × 250 mm, 5 μm) with acetonitrile-0.1%acetic acid solution as the mobile phase by gradient elution, and the detection wavelength was 260 nm. High performance liquid chromatography-diode array detector ( HPLC-DAD) for the determination of formononetin content was performed on AcclaimTM 120-C18 column ( 4.6 mm × 250 mm, 5 μm) with acetonitrile-water solution by isocratic elution, the detection wavelength was 254 nm, the flow rate was 1.0 mL/min and the column temperature was 25℃. Results The standard fingerprint of Caulis Spatholobi was set up through the evaluation of the fingerprints of 24 batches of Caulis Spatholobi samples from different habitats. Thirteen common peaks were identified with reference to formononetin peak, and the content of formononetin was determined by HPLC-DAD method. The similarity of the fingerprints of Caulis Spatholobi from different habitats and their formononetin content had great differences. Conclusion The established method is simple, accurate, highly sensitive, and repeatable, and can be applied for the quality control of Caulis Spatholobi.