Objective To study the differentiation and proliferation ability of the spinal neural stem cells (NSCs) at different gestational ages in fetal rats. Methods Sprague-Dawley fetal rats were divided into group A (12 days of pregnancy), group B (14 days of pregnancy) and group C (16 days of pregnancy). NSCs were separated with enzyme-assisted microdissection. The diameter and numbers of NSCs balls were measured at different time. The cell growth curve was drawn with CCK8 colorimeter. NSCs were identified with BrdU/Nestin immunohistochemical staining. They were induced with 10% fetal bovine serum for 10 days, and the expression of β-tubulinⅢ and glial fibrillary acidic protein was detected with immunocytochemistry. Results There were cells expressed BrdU, Nestin, β-tubulinⅢ and GFAP in all the group. The most cells (22.74±0.79%) expressed β-tubulinⅢ in the group B, but no significant difference between group B and group C. The cell vitality on the 5th day of third-generation neural stem cells was the most in group B. Conclusion For enzyme-assisted microdissection, it may obtain more neurons to isolate the neural stem cells from 14 days of pregnancy pregnant rats.

BACKGROUND: Considerable debate exists which factors influence the outcomes of the vitrified frozen-thawed embryo transfer because lack of clinical applications.OBJECTIVE: To explore the factors influencing the outcomes of the vitrified frozen-thawed embryo transfer in assisted reproductive technology.METHODS: A retrospective statistical analysis was performed in Reproductive Medicine Center, Renmin Hospital of Wuhan University of 142 patients, 154 thawing cycles. The patients were grouped according to the age of patients, embryonic development, fertilization methods, endometrial preparation programs, the endometrial thickness, the process of transplantation and the survival cell ratio of embryo recovery, the implantation rate and clinical pregnancy rate were compared between various groups.RESULTS AND CONCLUSION: Among the groups of the different age, fertilization methods, endometrial preparation programs, endometrial thickness and the process of embryo transfer embryo implantation rate and clinical pregnancy rate were no significant difference (P > 0.05); between the two groups of the second day fertilized embryos (D2) and the third day (D3)fertilized embryos, the clinical pregnancy rate was not significant (P> 0.05), but the embryo implantation rate of D3 group was significantly higher than D2 group. The survival cell ratio of embryo recovery has a significant effect on implantation rate and clinical pregnancy rate (P < 0.05). Ln the frozen-thawed embryo transfer cycles, embryo quality plays a major role in pregnancy rate, and preparation for appropriate endometrial thickness can improve the clinical pregnancy rate.

Aim The expression of NF-?B activation and the effect of antioxidant (N- acetylcysteine, NAC) on NF-?B activity in human airway epithelial cell was assessed. Methods Using the TNF-?,the airway epithelial cell strains of normal subject(16HBE) and tumor patient (H292) was activated and using Western-Blot and ELISA the expression of NF-?B and IL-8 were detected. Results It was found that the activity of NF-?B could be stimulated by the TNF-? and increase with the amount of TNF-? with the peak occurring at 2 to 4 hours after stimulation and then decreasing at six hours. At the same time, the level of IL-8 was elevated, but decreased with inhibition of NF-?B activity by NAC, that means the action of NAC has a dose-dependent effect. Conclusion NAC not only blocks the signal transmission activated by NF-?B, but also anticipates the transcription modulation of expression of many cell factors and inflammatory mediums. It suggests that NAC may play a role in the anti-inflammatory treatment of respiratory diseases.

Objective To study the present situation of benign prostatic hyperplasia (BPH) of male patients in geriatrics department. Methods Two thousand one hundred and twenty-five patients aged 60 years and above with BPH in geriatrics department of Chengdu, Lanzhou, Hangzhou, Wuhan, Changsha, Guangzhou, Shenyang, Shanghai, Nanjing, Jinan and Beijing were investigated using stratified cluster random sampling method. Results The constituent ratio of patients with benign prostatic hyperplasia in geriatrics department was 47.0%, and the proportions of the elderly of 60～,70～ ,80～ years old among the patients with benign prostatic hyperplasia were 18.1%,49.4 and 32.5%, respectively (x2 = 83.68, P<0.001). There was a significant difference in the occupation distribution of BPH constituent ratio: 66.4% for staffs of state organs, 18.0% for professional technical personnel and 14.7% for works and peasants (x2=410.27,P<0.001). The international prostate symptom scores (IPSS) were different among different cities (x2=101.00, P<0. 001 ). Logistic regression analysis showed that the IPSS was correlated with age and profession (age: OR= 1.029;worker and peasant as control group, working staff of state organs OR=O.707, professional technical personnel OR=0.339 ). Conclusions The benign prostatic hyperplasia is a common disease in male elderly patients in China, but the consultation rate is low. The IPSS of benign prostatic hyperplasia is correlated with age and profession. Prevention and treatment of benign prostatic hyperplasia in the elderly should be strengthened.

OBJECTIVE: To probe into the pharmacological mechanism of the Chinese medicinal compound Qinggong Zhixue Granule (QGZXG), which has an effect of replenishing qi to activate blood and expelling stasis to stop bleeding, in treating irregular vaginal bleeding after medical abortion. METHODS: Healthy female KM mice and mice with immunodeficiency caused by cytoxan (CTX) were chosen as experimental subjects. The effects of QGZXG on the phagocytization of the mice's monocytes (by using the method of carbon particles expurgation), the delayed type hypersensitivity caused by 2,4-dinitrochlorobenzene (DNCB) and the content of antibody of hemolysin in the mice's serum were observed. RESULTS: (1) QGZXG improved the ability of the liver and spleen of immunodeficient mice to expurgate carbon particles. There was an significant statistical difference between the CTX treated group and the low dose group as well as the high dose group (P < 0.01) and a statistical difference between the CTX treated group and the medium dose group (P < 0.05). (2) QGZXG boosted the cellular immunity of immunodeficient mice. The ear swelling of mice in low, medium and high dose group was more obvious than that in the CTX treated group, and this difference was significant in statistics (P < 0.01). (3) QGZXG raised the content of the antibody of hemolysin in the serum of immunodeficient mice. A statistical difference occurred between the high dose group and the CTX treated group. The low and medium dose also had a tendency of such effect. CONCLUSION: QGZXG can improve both the specific and non-specific immunity of the immunodeficient mice, and therefore accelerate the recovery of the whole body and endometrium after parturition and abortion. In consequence, it leads to shorter duration and less quantity of vaginal bleeding after medical abortion.

Hydrogenases are enzymes that catalyse the oxidation of hydrogen and the reduction of protons. It plays an important role in the process of biohydrogen production. According to the metal atoms within hydrogenase, it can be classified as NiFe-hydrogenase, Fe-hydrogenase and metal-free hydrogenase. The overwhelming majority of hydrogenases are metalloenzymes. The metal atoms are involved in the forming of active site and [Fe-S] clusters. The active site directly catalyzes the reduction of protons and the oxidation of hydrogen. The [Fe-S] clusters are involved in the transport of electrons between the H2-activating site and the redox partners of hydrogenase. Presently, the crystal structures of NiFe-hydrogenase and Fe-hydrogenase from a few kinds of microorganism have been revealed. The metal-free hydrogenase, characterized by the absence of [Fe-S] cluster and the presence of an iron-containing cofactor, shows a great diversity comparing with those of NiFe-hydrogenases and Fe-hydrogenases. Recent progress have also indicated the mechanisms of activation.
Catalysis ; Catalytic Domain ; Hydrogenase ; metabolism ; Iron-Sulfur Proteins ; metabolism ; Oxidation-Reduction

Objective To explore the learning and memory impairment and pathology in hippocampus in rats after spinal cord contu-sion. Methods 36 adult female Sprague-Dawley rats were randomly divided into sham group (n=18) and spinal cord injury group (n=18). Spinal cord injury model at T10 was established with modified Allen's technique (10 g × 25 mm). The hindlimbs behavior of rats was rated with Basso-Beattie-Bresnahan (BBB) scores once a week for 5 weeks. They were tested with motor evoked potentials (MEP) and Morris wa-ter maze 5 weeks after injury. The pathology of hippocampus was detected with HE staining 1 week, 3 weeks and 5 weeks after injury, 4 rats in a group, repectively. Results The BBB scores were significantly lower in the spinal cord injury group than in the sham group at each time point (P<0.05). The latencies of both N1 and P1 wave of MEP were significantly longer in the spinal cord injury group than in the sham group (P<0.001), while the amplitudes were significantly less (P<0.001). For the Morris water maze, the latency of arrival platform were sig-nificantly longer in the spinal cord injury group than in the sham group (P<0.001), and the time in target was significantly less (P<0.001), with more systematic positioning or annular positioning, while the sham group with more space-based positioning. Morphologically abnor-mal cells in hippocampus gradually increased since the first week after injury, with the decrease of cells survival, while it was normal in the sham group. Conclusion Spinal cord contusion can cause learning and memory impairment in rats, which may be related to injury in hippo-campus.

OBJECTIVETo study the expression level and significance of glucose transporter 1 (Glut-1) in normal breast tissue, adenosis, adenoma and breast carcinoma.

METHODSA total of 147 cases of female breast tissue samples, including 92 cases of invasive ductal carcinoma, 26 cases of breast fibroadenoma, 24 cases of breast adenosis and 5 cases of normal breast tissues, were collected for quantitative detection of the expression of Glut-1 protein by immunohistochemistry (EnVision method) and Western blot, and its mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSIn normal breast tissue and benign lesions of the breast, Glut-1 was undetectable or only weakly detectable in cytoplasm of ductal and acinar epithelia. In contrast, the intensity of Glut-1 staining was significantly higher in invasive ductal carcinomas (P = 0.0002) with protein expression predominantly in cellular membrane and lesser in cytoplasm. Western blot and RT-PCR analyses showed that the expression of Glut-1 protein and mRNA were significantly increased in invasive ductal carcinoma than fibroadenoma (P =0.001 for protein; P <0.05 for mRNA) and adenosis (P =0.001 for protein; P < 0.05 for mRNA). There was a significant difference among groups (P = 0.0002 for protein; P = 0.0001 for mRNA).

CONCLUSIONSGlucose transport activity, as indicated by Glut-1 protein and its mRNA expression, significantly increases in breast carcinoma than non-cancerous lesions. The over-expression of Glut-1 in breast carcinoma is tightly coupled with tumor cell proliferation, invasion and metastasis, implying that Glut-1 may serve as a new marker in the early diagnosis and prognostication of breast malignancy as well as a new therapeutic target.

Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Glucose ; physiology ; Glucose Transport Proteins, Facilitative ; genetics ; metabolism ; Glucose Transporter Type 1 ; genetics ; metabolism ; Humans ; Prognosis

BACKGROUNDC-reactive protein (CRP) has been reported to damage the vascular wall by inducing endothelial dysfunction and inflammation, and it is also speculated to have a role in attenuating angiogenic functions of human endothelial progenitor cells (EPCs). Interleukin-8 (IL-8) is an important mediator of the paracrine mitogenic effect of EPCs, which has direct angiogenic effects on mature endothelial cells. We, herein, investigated the direct effect of CRP on IL-8 production and gene expression in cultured human EPCs.

METHODSEPCs were isolated from the peripheral venous blood of healthy male volunteers. Cells were cultured in EndoCult liquid medium in the absence and presence of CRP at clinically relevant concentrations (5 to 25 microg/ml) for different durations (3 to 48 hours). IL-8 protein and mRNA of cultured EPCs were evaluated using ELISA and real-time PCR.

RESULTSThe results showed that CRP at a concentration of 10 microg/ml significantly reduced IL-8 secretion of cultured EPCs with a peak at 25 microg/ml, and also decreased mRNA expression in EPCs with a peak at 12 hours. In addition, preincubation of EPCs with SB203580, an inhibitor of p38 mitogen-activated protein kinase (MAPK) decreased CRP inhibition of IL-8 mRNA expression at 12 hours in EPCs.

CONCLUSIONSOur study, for the first time, demonstrates that CRP directly inhibits EPCs IL-8 secretion, a key cytokine player of angiogenesis induced by EPCs. Inhibition occurred in part via an effect of CRP to active the p38 MAPK signal transduction pathway in EPC. The ability of CRP to inhibit EPCs IL-8 secretion may represent an important mechanism that further links inflammation to cardiovascular disease.

C-Reactive Protein ; pharmacology ; Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Imidazoles ; pharmacology ; Interleukin-8 ; genetics ; Male ; Pyridines ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; drug effects ; metabolism ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

Based on the deletion information of high virulent PRRSV genome, 3 oligonucleotide primer were designed and synthesized. Specific and sensitive reverse transcription-PCR (RT-PCR) assays were de-veloped for the detection of high virulent PRRSV. The sensitivity and specificity of RT-PCR assays were evaluated, the results showing that the detection limit of the assay was found to be 0.265 pg of tissue total RNA, and the protocol have no cross-reaction with classical swine fever virus, porcine circovirus type 2,pseudorabies virus, streptococcus, haemophilus parasuis and Escherichia coli. Then 36 cell cultures, two PRRSV live vaccine strains and 184 clinical specimens from 52 farms were tested. Five PRRSV field iso-lates were the high virulent PRRSV; two PRRSV live vaccine strains from normal PRRSV, and 123 speci-mens from 42 farmer were positive (only 1 specimen was normal PRRSV). This RT-PCR method proved to be accurate differential diagnosis of the high virulent PRRSV and normal PRRSV with the characteristics of rapidity, sensitivity and specificity, and has a strong clinical relevance.