AbstractBACKGROUND: Fas and P53 are important regulator and control gene which can promote apoptosis. They belong to the receptor family part of tumor necrotic factor/nerve growth factor. Their expression products have effects on apoptosis signal transmission, and can regulate and control cell apoptosis in cerebral ischemia-reperfusion injury. And puerarin can alleviate the level of cell apoptosis.OBJECTIVE: To observe the effect of puerarin on Fas and P53, the apoptosis-related gene of nerve cell in hippocampns CA1 region of rats af ter cerebral resuscitation.DESIGN: Randomized controlled trial. SETTING: Department of Emergency, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: The experiment was carried out at Emergency Department, Tongji Hospital, Tongji Medical College, Huazhong University ofScience and Technology from September 2001 to Februray 2002. Totally 45 of 3 months old Wistar rats of clean grade were selected, and randomly divided into 3 groups: sham operation group, model control group and puerarin treatment group with 15 rats in each group.METHODS: Acute global brain ischemia-reperfusion models were established in rats of puerarin treatment group and model control group. In rats of sham operation group, stigmata of both flanks of the first cervical vertebrae were isolated, but bilateral vertebral arteries were not electric coagulated, and biolateral common carotid arteries were only isolatedwithout clamping close. Rats in puerarin treatment group were given puerarin injection 100 mg/kg, 1 hour before ischemia, and model control group were given normal saline in equivalence while rats in sham operation group were not given medicine. Death of rats in each group was performed separately in the 3rd, 6th, 12th, 24th and 48th hours after cerebral ischemia-reperfusion with 3 rats per group in each time. Hippocampus tissues of rats were isolated, and tissue slices were preparated. And the changes of- the protein expression levels and the number of apoptosis cells of rats in each group at different time point after cerebral ischemia-reperfusion were detected in immuno-histochemical method and end labelling in situ method. MAIN OUTCOME MEASURES: ① The number of positive cells inprotein expression of Fas and P53 in hippocampus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion was studied. ② Comparison of the number of apoptosis cells in hippocampus CA1 region of rats between groups at different time point after cerebral ischemia-reperfusion were studied, too.RESULTS: All the 45 rats enrolled in research were entered the stage of result analysis: ① The number of positive cells in protein expression of Fas in hippocampus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion: Obvious gene expression of Fas was not found in sham operation group. In contrast with model control group, obvious decrease was found at all time points after cerebral ischemi a-reperfusion in puerarin treatment group, and in the 6th, 12th, 24th and 48th hour the differences were significant [(15.0±4.3), (13.5±4.9); (40.7±3.4), (27.2±3.1); (37.0±4.8), (22.0±2.1); (24.7±4.1), (18.9±5.3)/mm; P < 0.05,P < 0.01]. ② The number of positive cells in protein expression of P53 in hippocanpus CA1 region of rats in each group at different time point after cerebral ischemia-reperfusion: Obvious gene expression of P53 was not found in sham operation group. In contrast with model control group, obvious decrease was found in the 24th and 48th hour after cerebral ischemiareperfusion in puerarin treatment group [(25.3±4.4), (12.8±2.7); (24.3±3.6), (10.9±3.0)/mm; P < 0.01]. ③ Comparison of the number of apoptosis cells in hippocampus CA1 region of rats between groups at different time point after cerebral ischemia-reperfusion :In contrast with model control group,obvious decrease was found in the 12th, 24th and 48th hour after cerebral is chemia-reperfusion in puerarin treatment group [(34.0±3.7), (21.0±3.7); (41.0±4.2), (33.0±4.8); (71.0±5.5), (41.0±3.4)/mm; P < 0.01].CONCLUSION: In rats which were given puerarin treatment, the expression of Fas decrease obviously in 6 to 48 hours after cerebral ischemiareperfusion, and the expression of P53 decreased obviously in the 24th to 48th hour after cerebral ischemia-reperfusion, and a descent tendency could be found in the number of apoptosis cells. These can further prove the cerebral protective effect of puerarin, and indicate that the inhibition of puerarin to cell apotosis after cerebral resuscitation is related to its effect on the decrease in protein expression of apoptosis-promoting gene, Fas and P53.Puerarin has a protective effect on cerebral ischemia-reperfusion injury of rats. In comparison with model control group, the expression of Fas in puerarin treatment group has an obvious decrease inthe 6th to 48th hour after cerebral ischemia-reperfusion, the expression of P53 has an obvious decrease in the 24th to 48th hour after cerebral ischemia-reperfusion, and the number of apoptosis cells decrease obviously, too, which further improves the cerebral protective effect of puerarin and indicates that the inhibition of puerarin to cell apoptosis after cerebral resuscitation is related to its effect on the decrease in protein expression of apoptosis-promoting gene Fas and P53.

Objective To study the model of separation and culture and the growth characteristics and osteogenic capability of bone marrow stromal stem cells (BMSCs). Methods BMSCs were isolated from adult rats using gradient centrifugation and anchoring culture. The passage cells were induced to differentiate into osteoblasts by means of an differentiation induction medium containing dexamethasone,?-glycerophophate and Vitamin C. Alkaline phosphatase (ALP) activity and the ability to form mineralized nodules were examined. Results Colonies of stromal stem cells were formed in the primary culture and contacted one another at the 14th day. In the passage culture, cells became bigger than those in the primary culture and could be subcultured one generation in 5～7 days. After the culture in the differentiation induction medium, the ALP activity was enhanced significantly and the mineralized nodules were formed. Conclusion BMSCs can be obtained easily and have strong proliferation ability and osteogenic capacity. This study suggests that BMSCs can become seeding-cells in bone tissue engineering.

ObjectiveTo investigate the clinical effects of posterior atlantoaxial fusion fixation in treatment of old atlantoaxial injury secondary to atlantoaxial dislocation.MethodsA retrospective analysis was carried out on 16 patients ( 14 males and 2 females) with old atlantoaxial injuries secondary to atlantoaxial dislocations managed with posterior atlantoaxial fusion fixation from March 2008 to March 2012.The time from injury to operation lasted for 3-36 months ( average 10.5 months).Posterior atlantoaxial transpedicular fixation was performed in 13 patients including 10 patients with old odontoid fractures and three with old traumatic transverse ligament disruptions of the atlas combined with atlantoaxial dislocations.Also,posterior atlantal arch transpediclar fixation combined with axial pedicle screw fixation was performed in three patients who had old odontoid fractures combined with atlantoaxial dislocations.All patients had simultaneous autogenous bone grafting between atlas and axis during reduction and fixation.The preoperative and postoperative Japanese Orthopaedic Association (JOA) scores were compared.Follow-up X-ray films and CT was performed to evaluate the atlantoaxial reduction and fusion.ResultsAll the patients were followed up for 9-18 months ( mean 13 months).None of the patients had spinal cord or vertebral artery injuries.Follow-up CT showed that two patients had partial penetration of one side axial pedicle screws into transverse foramen without nerve and blood vessel injuries.Clinical symptoms obtained different degree of improvement.The postoperative JOA scores ranged from 13 to 16 points ( mean 14.8 points) and the improvement rate of JOA was 71％-92％ ( mean 82％ ).The X-ray films and CT showed sound bone fusion,with good location of screws but with no signs of atlantoaxial instability or loss of reduction,or loosening or breakage of the screws.ConclusionPosterior atlantoaxial fusion fixation can effectively reconstruct atlantoaxial stability,improves neurologic function of spinal cord and has reliable curative effects.

The role of serum and glucocorticoid-induced kinase 1 (SGK1) pathway in the connective tissue growth factor (CTGF) expression was investigated in cultured human mesangial cells (HMCs) under high glucose. By using RT-PCR and Western blot, the effect of SGK1 on the CTGF expression in HMCs under high glucose was examined. Overexpression of active SGK1 in HMCs transfected with pIRES2-EGFP-S422D hSGK1 (SD) could increase the expression of phosphorylated SGK1 and CTGF as compared with HMCs groups transfected with pIRES2-EGFP (FP) under high glucose or normal glucose. Overexpression of inactive SGK1 in HMCs transfected with pIRES2-EGFP-K127N hSGK1 (KN) could decrease phosphorylated SGK1 and CTGF expression as compared with HMCs groups transfected with FP under high glucose. In conclusion, these results suggest that high glucose-induced CTGF expression is mediated through the active SGK1 in HMCs.
Cells, Cultured ; Connective Tissue Growth Factor/genetics ; Connective Tissue Growth Factor/*metabolism ; Glucose/*pharmacology ; Immediate-Early Proteins/metabolism ; Immediate-Early Proteins/*physiology ; Mesangial Cells/cytology ; Mesangial Cells/*metabolism ; Protein-Serine-Threonine Kinases/metabolism ; Protein-Serine-Threonine Kinases/*physiology ; Signal Transduction/drug effects

Humans ; Liver Cirrhosis ; Renin-Angiotensin System ; Self-Control

Summary: The role of serum and glucocorticoid-induced kinase 1 (SGK1) pathway in the connective tissue growth factor (CTGF) expression was investigated in cultured human mesangial cells (HMCs) under high glucose. By using RT-PCR and Western blot, the effect of SGK1 on the CTGF expression in HMCs under high glucose was examined. Overexpression of active SGK1 in HMCs transfected with pIRES2-EGFP-S422D hSGK1 (SD) could increase the expression of phosphorylated SGK1 and CTGF as compared with HMCs groups transfected with pIRES2-EGFP (FP) under high glucose or normal glucose. Overexpression of inactive SGK1 in HMCs transfected with pIRES2-EGFP-K127N hSGK1 (KN) could decrease phosphorylated SGK1 and CTGF expression as compared with HMCs groups transfected with FP under high glucose. In conclusion, these results suggest that high glucose-induced CTGF expression is mediated through the active SGK1 in HMCs.

OBJECTIVETo investigate the effect of dracorhodin perchlorate (DP) on inhibiting high glucose-induced serum and glucocorticoid induced protein kinase 1 (SGK1) and fibronectin (FN) expression in human mesangial cells (HMC), and its mechanism of prevention and treatment on renal fibrosis in diabetic nephropathy (DN) .

METHODThe HMC were divided into normal glucose group (NG group, 5.5 mmol x L(-1) D-glucose), normal glucose +low DP group (NG + LDP group, 5.5 mmol x L(-1) D-glucose +7.5 micromol x L(-1) DP), normal glucose +high DP group (NG + HDP group, 5.5 mmol x L(-1) D-glucose + 15 micromol x L(-1) DP), high glucose group (HG group,25 mmol x L(-1) D-glucose), high glucose +low DP group (HG + LDP group, 25 mmol x L(-1) D-glucose + 7.5 micromol x L(-1) DP)and high glucose +high DP group (HG +HDP group, 25 mmol x L(-1) D-glucose + 15 micromol x L(-1) DP). Each group was examined at 24 hours. The levels of SGK1 and FN mRNA was detected by real-time fluorescence quantitative PCR,and the expression of SGK1 and FN protein was detected by Western blot or indirect immunofluorescence.

RESULTA basal level of SGK1 and FN in HMC were detected in NG group, and the level of SGK1 and FN mRNA and protein were not evidently different compared to that of NG group adding 7.5 micromol x L(-1) DP for 24 hours. On the other hand, the levels of SGK1 and FN mRNA and protein were obviously decreased by adding 15 micromol x L(-1) DP for 24 hours. Compared to NG group, the levels of SGK1 and FN mRNA and protein were increased in HG group after stimulating for 24 hours (P < 0.01). Compared to HG group, the level of SGK1 and FN mRNA and protein were evidently reduced in HG + LDP and HG + HDP groups by adding 7.5 micromol x L(-1) DP and 15 micromol x L(-1) DP for 24 hours (P < 0.01).

CONCLUSIONDracorhodin perchlorate can inhibit high glucose-induced serum and glucocorticoid induced protein kinase 1 (SGK1) and fibronectin(FN) expression in human mesangial cells, and this may be part of the mechanism of preventing and treating renal fibrosis of DN.

Benzopyrans ; pharmacology ; Cell Line ; Diabetic Nephropathies ; drug therapy ; enzymology ; genetics ; metabolism ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibronectins ; biosynthesis ; genetics ; Gene Expression ; drug effects ; Glucose ; metabolism ; Humans ; Immediate-Early Proteins ; genetics ; metabolism ; Mesangial Cells ; drug effects ; enzymology ; metabolism ; Perchlorates ; pharmacology ; Protein-Serine-Threonine Kinases ; genetics ; metabolism

Since the outbreak of corona virus disease 2019(COVID 19), the epidemic has spread rapidly, which brings great challenge to the surgical diagnosis, treatment and management of lung neoplasm Sichuan International Medical Exchange &Promotion Association organized thoracic surgery experts to sum up experiences from experts in major hospital, and formulated the Guidance suggestion on surgical diagnosis, treatment and management of lung neoplasm during the outbreak of COVID-19 to provide references for thoracic surgeons.