Teaching method is important to carry through the content of the course and guarantee the teaching quality.We have proceeded some reforms and research in the Obstetrics and Gynecology teaching and have obtained the good result by intergroting teaching conteut and teaching resoures,selecting high-level teaching team,case teaching,PBL teaching and multi-media teaching.

Objective To evaluate the value of combined use of laparoscopy and hysteroscopy in the diagnosis of infertility. Methods Clinical data of 168 cases of infertility receiving examinations with laparoscopy and hysteroscopy from June 1999 to October 2003 were retrospectively reviewed.Results Hysteroscopic examinations found intrauterine diseases in 79 cases(79/168,47.0%),including 46 cases of endometrial hyperplasia or polyps(46/79,58.2%).Laparoscopic examinations showed organic pelvic diseases in 99 cases,including 85 cases of chronic pelvic inflammation,endometriosis or polycystic ovarian syndrome(85/99,85.9%).Both laparoscopy and hysteroscopy gave normal findings in 15 cases and abnormal findings in 39 cases.Unilateral or bilateral tubal obstruction was found in 90 cases by tubal patency tests under hysteroscope(90/168,53.6%) and in 78 cases by laparoscopy(78/168,46.4%). Conclusions Combined use of laparoscopy and hysteroscopy offers accurate diagnostic evidences in examinations of infertility.

Objective The purpose of this study was to explore the teaching method PBL in gynecology and obstetric education. Methods We took PBL in teaching Acute Gynecological Abdomen Pain in 102 students between 2008 and 2009. Results Questionnaire results and high scores in tests suggested that the students showed great subjective initiative in studies.PBL can cultivate students'self-learning capability,improve their ability of analyzing and solving prob-lems. Conclusion PBL is a better way in clinical medical education and worthwhile to be further researched and carried out.

Objective To investigate the role of p38 MAPK pathway in the expression of high mobility group box 1 (HMGB1) in lung tissue in a rat model of ventilator-induced lung injury. Method Twenty-fonr healthy Sprague Dawley (SD) rats were randomly divided into 3 groups (n = 8 each) : group A, spontaneous breathing; group B, small tidal volume ventilation (Vt = 8 mL/kg) and group C, high tidal volume ventilation (Vt = 40 mL/kg). 1he animals in group B and C were mechanically ventilated for 4 hours and all animals were sacri-riced. The lungs were removed for: (1) lung lavage and determination of total protein contnt and WBC and neu-trophil counts in broncho-alveolar lavage fluid (BALF) ; (2) determination of W/D lung weight ratio and myelop-erexidnse (MPO) activity; (3) detennination of HMGB1 protein and mRNA expression and p38 MAPK activity in lung tissue. Differences within the groups were analyzed using One way ANOVA. Results The inflammatory re-sponse as evidenced by total protein (1.77 ± 0.68) g/L and WBC (106.55 ± 28.17) × 10~7/L in BALF, W/D lung weight ratio (7.16±1.02) and MPO activity (3.94±1.21) U/g were significantly higher in group C com-pared with group A (P <0.05); HMGB1 protein (0.64±0.17) and mRNA (1.17±0.45) expression and p38 activity (0.51±0.12) also significantly increased in group C (P <0.05). Of the above indexes, there were no statistical differences between group B and group A (P > 0.05). Conclusions High tidal volume ventilation in-daces acute lung injury, which may be related with upregulation of HMGB1 expression through p38 MAPK signal pathway.

AIM: To screen the binding proteins to HMGB1 promoter by phage display technique. METHODS: HMGB1 promoter was incubated with phage display library. Unbound phages were eluted and phages bound to HMGB1 promoter were amplified. Twenty individual clones were randomly selected and identified by enzyme-linked immunosorbent assay (ELISA). Positive clones were characterized by DNA sequencing and the sequences were subjected for computer analysis. RESULTS: Positive phages binding to HMGB1 promoter were enriched after 4 rounds of biopanning. Twenty phage clones were selected and eleven clones of which were identified to bind specifically to HMGB1 promoter. The sequences in full length were obtained and searched for homologous sequences from GenBank. Altogether eight coding sequences were obtained, six of which were known proteins including activator protein-1(AP-1) and two of which were uncharacterized ones. CONCLUSION: Several proteins were obtained that bind specifically with HMGB1 promoter. The results will be useful for further studying the expression and regulation mechanism of HMGB1.

Objective To investigate the role of extracellular regulated protein kinase (ERK) signal pathway in mechanical stretch induced high mobility group box 1 protein (HMGB1) expression on alveolar epithelial cells (A549). MethodsA549 cells were cultured and seeded at 1×10~5 cells/ml in 6-well Bioflex cell culture plates. Subsequently, the cells were exposed to cyclic mechanical stretch at 14% (group B) elongation for 4 hours using Flexercell 4000T cell stretching unit. In group C, cells were pretreated with PD98059 for 2 hours before mechanical stretch. Cells in group A without stretch were served as control. The expression of HMGB1 protein and mRNA in A549 cells were detected by immunocytochemisty staining and RT-PCR, respectively. ERK activity was measured by Western blotting method. Results Immunocytochemisty staining indicated that the expression of HMGB1 protein in A549 cells was increased obviously in group B (P<0.05) and decreased in group C (P<0.05). Polymerase chain reaction (RT-PCR) showed that the expression of HMGB1 mRNA was also significantly increased in group B (P<0.05) and decreased in group C (P<0.05). Western blotting analysis confirmed the activation of ERK in A549 cells by mechanical stretch (P<0.05). PD98059, an inhibitor of ERK, might significantly inhibit mechanical stretch induced HMGB1 protein and mRNA expression in A549 cells (P<0.05). Conclusion Mechanical stretch could regulate the expression of HMGB1 gene and protein in A549 cells through ERK signal pathway.

Aim To examine the effect of different dosages hydroxyethyl starch(HES)130/0.4 on intercellular adhesion molecule 1(ICAM-1)expression in lung tissue of acute lung injury in endotoxemic rats and explore the role of MAPKs pathway in its expression.Methods Thirty six healthy Sprague Dawley(SD)rats weighing 270~320 g were randomly divided into 6 groups with 6 animals in each group.In group H1-H4,1 min after lipopolysaccharide(LPS)5 mg?kg-1 intravenously administration,HES 130/0.4 with 3.75,7.5,15,30 ml?kg-1 were infused intravenously respectively at a rate of 0.2 ml?min-1.In group L,saline instead of HES 130/0.4 was administered.Group N served as control by giving the same volume of saline.The animals were anesthetized with pentobarbital.Right external jugular vein was injected with LPS.The animals were killed 4 hours after LPS injection for determination of total protein,WBC,MPO,W/D,ICAM-1 protein and mRNA and MAPKs activity.Results Compared with control group,total protein,WBC,MPO,W/D,expression of ICAM-1 protein and mRNA and MAPKs activity were increased significantly in group L.Compared with group L,total protein,WBC,MPO,W/D,expression of ICAM-1 protein and mRNA and MAPKs activity were decreased significantly in group H1 and H2,especially at the dosage of 7.5 ml?kg-1.Conclusion HES 130/0.4(7.5 ml?kg-1)can attenuate inflammatory response of acute lung injury induced by LPS,which may be related with inhibiting the expression of ICAM-1 protein and mRNA through MAPKs signal pathway.

Objective To construct the red fluorescent protein reporter gene vector containing high mobility group box 1 protein(HMGB1) promoter sequence and study the regulation mechanism of the expression of HMGB1gene under mechanical stretch.Methods HMGB1 promoter was subcloned into a red fluorescent protein vector,pDsRed1-1.After identified by PCR,enzyme digestion and DNA sequencing,the recombinant vector pDsRed1-1-HMGB1P was then transfected into HEK293 cells.Blank vector or pDsRed-1 was transfected into 293 cells and served as controls.The expression of red fluorescent protein and its reaction to mechanical stretch were observed under a fluorescent microscope.HEK293 cells transfected with pDsRed1-1 vector served as control.Results PCR,double restriction enzyme digestion and DNA sequence analysis showed that the recombinant vector,pDsRed1-1-HMGB1P,was constructed correctly.This vector was lowly expressed in HEK293 cells of resting state.But after stimulated by mechanical stretch,strong red fluorescence was observed.No red fluorescence was observed in the control cells.Conclusion A red fluorescent protein reporter gene vector containing HMGB1 promoter sequence has been constructed successfully and expressed highly in mammalian cells.Since it responds to mechanical stretch effectively,it can thus provide a convenient tool to study the regulation mechanism of the expression of HMGB1 gene by mechanical stress.

Objective To evaluate the relationship between 25-(OH) vitamin D [25-(OH) D] level and peripheral neuropathy in patients with type 2 diabetes mellitus.Methods Eighty patients with type 2 diabetes mellitus were enrolled in this cross-sectional study,including 37 subjects with and 43 without diabetic neuropathy.Anthropometric data was collected and serum levels of 25-(OH) D,HbA1c,blood lipid,and hepatic and renal functions were determined in all patients.Results Serum 25-(OH) D level was significantly lower in patients with diabetic neuropathy compared to those without neuropathy [(12.73 ± 4.68 vs 17.56 ± 5.28) ng/ml,P＜0.01].Logistic regressions demonstrated that vitamin D level was associated with diabetic neuropathy (OR=1.222,95％ CI 1.095-1.364).Conclusions Vitamin D insufficiency is associated with diabetic peripheral neuropathy.25-(OH) D level seems to be an independent risk factor of diabetic neuropathy in patients with type 2 diabetes mellitus.

BACKGROUNDNeurofibromatosis type 1 (NF1) is the most common genetic syndrome predisposing patients to various tumors due to dysregulation of the Ras signaling pathway. Recent research has shown NF1 patients also suffer a spectrum of bone pathologies. The pathogenesis of NF1 bone diseases is largely unknown. There is no current treatment. By Nf1 heterozygote (Nf1+/-) mice and Nf1 conditional knockout mice, we and other groups demonstrated abnormal osteoblast and osteoclast function due to dysregulation of Ras signaling. However, the specific downstream effector pathways linked to NF1 abnormal osteoblastogenesis and osteoclastogenesis have not been defined. In this study, we investigated the Ras downstream effector related with NF1 bone disease.

METHODSWe used Nf1+/+ and Nf1+/- mice as normal and NF1 models. Bone stromal cells extracted from Nf1+/+ and Nf1+/- mice were induced osteoclasts. The osteoclast cell was stained by tartrate resistant acid phosphatase staining. The osteoclast cell number was counted and the surface area of osteoclast cells was calculated under the microscope. The mRNA of mammalian target of rapamycin (mTOR) was determined by quantitative reverse-transcription-polymerase chain reaction. The presence of ribosomal protein S6 kinase was determined by Western blotting.

RESULTSCompared with Nf1+/+ mice, Nf1+/- mice had about 20% more of osteoclast cells. These osteoclast cells were larger in size with more nuclei. Hyperactive mTOR was detected in Nf1+/- osteoclast cells. Inhibition of mTOR signaling by rapamycin in Nf1+/- osteoclasts abrogated abnormalities in cellular size and number.

CONCLUSIONmTOR pathway inhibition may represent a viable therapy for NF1 bone diseases.

Animals ; Male ; Mice ; Neurofibromatosis 1 ; drug therapy ; Osteoclasts ; drug effects ; physiology ; Osteogenesis ; drug effects ; Sirolimus ; pharmacology ; TOR Serine-Threonine Kinases ; antagonists & inhibitors ; physiology