Objective　To investigate the clinical results of the medial multiplex flap pedicled with the posterior tibial vessel. Methods　Twelve cases with soft tissue defects and bone defects of limbs were treated with the medial multiplex flap pedicled with the posterior tibial vessel from September 1992 to May 1999. Among them, bone and soft tissue defects following opened fracture in 7 cases, chronic ulcer following chronic osteomyelitis in 2 cases, melanoepithelioma in 2 cases, bone and soft tissue defects following osteoma resection in 1 case. The bone defect area was from 2.5 cm×5.0 cm to 4.5 cm ×11.0 cm. Free graft was performed in 5 cases, bridged transposition in 3 cases and reversal transposition in 4 cases, among them, periosteal myocutaneous flap with autogenous or allogeneic bone grafting in 8 cases, myocutaneous flap in 4 cases. The area of the flaps from 6 cm ×8 cm to 12 cm×25 cm. Results　All flaps were healed by first intention, but in the distal fragments of bigger flaps were partially necrosed in 2 cases. In 10 cases bone healing were obtained after 16 weeks of operation according to the X-ray photos. All cases were followed up from 6 to 18 months. All cases achieved satisfactory result but 1 case died because of lung metastasis of osteoma. Conclusion　The multiplex graft pedicled with the posterior tibial vessel is an ideal graft for repairing the large soft tissue defects and bone defects, because it has such advantages as adequate blood supply, big vascular diameter, long pedicle and big dermatomic area.

Objective To establish an effective method for inducing mouse bone marrow mononuclear cells(MNCs) to differentiate into hepatocytes.Methods MNCs were isolated by gradient density centrifugation,and the cells were cultured in Iscove's Modified Dulbecco's medium supplemented with 10 % new bovine serum(NBS),20 ng/ml hepatocyte growth factor(HGF),10 ng/ml fibroblast growth factor-4(FGF-4) and 10 ng/ml oncostatin m(OSM) for 21 days' induction.The medium was changed every 3 days.Results When MNCs were cultured with HGF,FGF-4 and OSM,cuboidal morphology was observed,and cells also expressed marker genes specific for liver cells in a time-dependent manner.?-fetoprotein(AFP) and cytokeratin 19(CK19) were expressed on the day 7,and CK18 and TAT were detected on the day 14-21,in common with the immunofluoresence results.Differentiated cells further demonstrated these cells also acquired functional characteristics of hepatocytes.Conclusion Mouse MNCs can differentiate into hepatocytes when induced by HGF,FGF-4 and OSM.

OBJECTIVE:To observe the clinical efficacy and safety of Calcitriol soft capsules combined with Telmisartan tab-lets in the treatment of early diabetic nephropathy(DN),and the effect on the levels of inflammatory factors. METHODS:Totally 110 patients with early DN were randomly divided into observation group and control group. The control group was orally given Telmisartan tablets with the initial dose of 40 mg,qd,and the maximum dose was 80 mg,qd;the observation group was orally giv-en Calcitriol soft capsules 0.25μg based on the treatment of control group,qd. The course was 1 month. The clinical data was com-pared,including the clinical efficacy and 24 h urinary protein,serum creatinine(Scr),urinary albumin excretion rate(UAER),se-rum C reactive protein (CRP),tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) before and after treatment. The adverse reactions were observed. RESULTS:After treatment,the total effective rate in observation group was significantly higher than con-trol group,with significant difference (P<0.05);the 24 h urinary protein,Scr,UAER,and levels of CRP,TNF-α and IL-6 in observation group were significantly lower than control group and before treatment,with significant differences(P<0.01 or P<0.05). There were no obvious adverse reactions in 2 groups. CONCLUSIONS:Calcitriol soft capsules combined with Telmisartan tablets has better efficacy than only Telmisartan tablets in the treatment of DN,and can more effectively improve the levels of CR, TNF-αand IL-6,which is helpful to delay progression of patients.

Objective To investigate therapeutic effects of Salvia miltiorrhiza on hepatic fibrosis of rats induced by carbon tewachloridean. Methods Rat models of hepatic fibrosis were founded by intraperitoneal injection of carbon tetrachloride. Salvia miltiorrhiza were given to these rats. Normal group and control group were set for comparison at the same time. Serum levels of ALT, AST, HA, LN, and PC-Ⅲ were detected; HE and Masson staining were conducted in hepatic tissues to observe pathological variations. Results Salvia miltiorrhiza could decrease serum levels of ALT, AST, HA, LN,PC-Ⅲ obviously (P ＜0.01), compared with the control group; Salvia miltiorrhiza could obviously improve pathological variations compared with the control group. Conclusion Salvia miltiorrhiza has therapeutic effect on hepatic fibrosis of rats

AIM: To determine the content of emodin and physcion in rat plasma by nonaqueous RP-HPLC. METHODS: After hydrolysis and extraction, the content of emodin in the plasma was determined by nonaqueous RP-HPLC. The separation was performed on Kromasil C 18 column (250 mm?4.6 mm, 5.0 ?m) with the mobile phase comprised of methanol-acetic acid (99.9∶0.1). The flow rate was 1.0 mL?min -1 and the detection wavelength was at 254 nm. RESULTS: The linear ranges for emodin and physcion were in the range of 0.0425-2.8 ?g?mL -1 and 0.0491-3.14 ?g?mL -1 , respectively. The average recoveries of emodin and physcion were 95.7%-100.1% and 96.2%-99.8%, with corresponding RSD of 1.3% and 1.6% respectively. CONCLUSION: This method is simple, rapid, accurate and reproducible with RP-HPLC to detect rhein in plasma.

Objective To investigate the effects of brain ischemic preconditioning (BIP) on peripheral blood EPCs and neovascularization in ischemic brain tissue in rats with cerebral ischemia reperfusion injury (IRI). Methods One hundred and eight male SD rats were randomly divided into three groups:SO group (n=36), MCAO group (n=36) and BIP group (n=36). Neurological function assessment was conducted at 0 h before MCAO-reperfusion, 3 h, 24 h and 3 d, 5 d as well as 7 d after MCAO-reperfusion (n=6 for each group in each time point). Flow cytometry was used to calculate the number of EPCs. Immunohistochemical staining was used to detect the capillary density. Results ①Although neurologi?cal deficit scores were significantly decreased in both BIP and MCAO groups after 3 h following MCAO-reperfusion, the scores were much lower in BIP group than in MCAO group(5 d:1.00±0.63;7 d:1.00±0.63, P<0.05).②The numbers of EPCs were decreased in MCAO group while was increased in BIP group at 3 h after MCAO-reperfusion. The numbers of EPCs were significantly higher in BIP group than in MCAO group(24 h:0.58±0.07;3 d:0.80±0.10;5 d:0.68±0.05;7 d:0.52 ± 0.03, P<0.01). ③ The new blood vessels could be detected at 3 d in BIP group and 5 d in MCAO group after MCAO-reperfusion. The numbers of new blood vessels were significantly higher in BIP group than MCAO group(5 d:14.53 ± 3.44; 7 d: 41.40 ± 5.62, P<0.01). ④ Pearson analysis showed a positive correlation between EPCs and capillary density (5 d: r=0.855, P<0.01; 7 d: r=0.946, P<0.01). Conclusion BIP can improve EPCs mobilization and function, which may contribute to neovascularization in the ischemic brain tissue.

Objective To examine loss of heterozygosity (LOH) and microsatellite instability (MSI) of locus D8S532 on chromosome 8 and their influence on the expression of sFRP1 in the hepatocellular carcinoma (HCCs), which may provide an experimental evidence for clarifying the mechanism of sFRP1 gene and tumor development. Methods DNA was extracted from formalin-fixed paraffin-embedded tissues. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and ordinary silver stain were used to study LOH and MSI of locus D8S532. Envision immunohistochemistry, Leica-Qwin computerized imaging system and Image-Pro PluS (IPP) version 4.5 professional imaging analysis software were used to assess the expression of sFRP1. Results The detection rates of LOH and MSI of locus D8S532 in the 36 specimens of HCC were 11.11% and 8.33% respectively. The down-regulation of sFRP1 was observed in 31 of 36 HCCs (86.11%) compared with non-carcinoma liver tissues, and the positive rate of sFRP1 protein of the HCCs was 52.78%( 19/36 ). The frequency of LOH was lower in the cases with positive expression of sFRP1 protein than those negative (0 vs 23.53%, P <0.05). Conclusion It was a common phenomenon that expression of sFRP1 protein is negative or low in Chinese with HCCs. The genetic instability of sFRP1 gene was one of causes, which lead to HCCs. LOH may play a major role in negative expression of sFRP1.

Objective To observe therapeutic effects of HuGanJieXian decoction on rats hepatic fibrosis induced by tetrachloride. Methods Rat models of hepatic fibrosis were constructed by intraperitoneal injection of tetrachloride.HuGanJieXian decoction composed of low, middle, and high dose curcumin were given to these rats respectively at the same time. Sho-saiko-to compound treatment group and Fufangbiejiarangan Tablets treatment group were made as positive control groups. After twelve weeks, all rats were executed. Serum samples were kept for measuring serum levels of PC-Ⅲ, LN, and HA. Left livers were extirpated for pathologic examination including H.E and Masson stainings. Grade of hepatic fibrosis were evaluated according to SSS system. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) of supematant centrifugated from hepatic tissue homogenate were detected. Results Serum levels of PC-Ⅲ, LN, and HA were depressed obviously in decoction groups compared with those of fibrotic group (P<0.05) , especially in the low-dose curcumin group.HuGanJieXian Decoction could increase the level of SOD and decrease the level of MDA (P<0.05) , especially in the low-dose curcumin group. Staining of H. E and Masson showed that degrees of hepatic fibrosis in decoction groups were improved obviously compared with that of the fibrotic group. Conclusion HuGanJieXian Decoction can improve rat hepatic fibrosis, the mechanism of this effect may be associated with protecting hepatic cell membrane and anti- peroxidative damage.

ObjectiveTo investigate the effect of NGF on apoptosis of HSC in vitro and explore the possible mechanism.MethodsHSC was incubated with different concentrations of NGF.HSC apoptosis was identified by FCM.The expressions of apoptosis-regulating proteins Caspase-3,p53 and Bcl-2 of HSC after apoptosis induced by NGF were examined by immunohistochemical staining.Expressions of NGF and p75NTR were detected by immunofluorescence.ResultsApoptosis index of HSC was higher than that of control group [(22.36±9.51)％ vs (5.88±1.36)％] after treated with NGF (100 ng/ml) (P＜0.05).After incubating with 100 ng/ml NGF for 24 h,the positive expression rates of p53 and Caspase-3 of HSC increased significantly than those of control group [(78.41±4.00)％ vs (34.96±3.84)％,(39.26±1.57)％ vs (9.27±1.01)％,P ＜0.05].The positive expression rate of Bcl-2 protein of HSC significantly decreased compared with that of control group (18.12±1.38)％ vs (91.53±2.98)％ (P＜0.05).When HSC was stimulated with 100 ng/ml NGF for 24 h,the average optical density of NGF increased significantly than control group (6.53±1.40 vs 1.77±0.17) (P＜0.05),while the expression of p75NTR was not significantly changed (3.52±0.36 vs 4.24±0.38) (P＞0.05).ConclusionsThe mechanism of NGF to induce HSC apoptosis may be associated with the up-regulating expression of Caspase3,P53 and down-regulating expression of Bcl-2 on HSC.NGF could be used as an initiating factor and effect factor to increase the expression of NGF on HSC,but it had no significant effect on p75NTR expression.

Objective To compare the changes in energy metabolism in 2-chloroethyl ethryl sulfide(CEES)-poisoned bronchial epithelial cell 16HBE cultured in media at different glucose concentrations .Methods Bronchial epithelial cell 16HBE was cultured in high (4.5 mg/ml) or low (1.1 mg/ml) glucose medium and exposed to a sulfur mustard simulant CEES of 0.2, 0.5, 1.0 mmol/L.Cell growth and cytotoxicity were tested using MTS .ATP, ADP and AMP were detected by HPLC and the value of ATP/ADP, total adenine nucleotides ( TAN) and energy charge ( EC) was subsequently calculat-ed.Mitochondrial oxidative phosphorylation-related proteins, COX-10 and ISCU, were detected using Western blotting . Rhodamine 123 was applied to detect the mitochondrial membrane potential using flow cytometry .Results Low glucose accelerated the growth and energy metabolism of 16HBE cells in regular culture , and the contens of ADP , TAN, COX-10 and ISCU in low glucose group were significantly higher than those in high glucose group .CEES exposure (≥0.5 mmol/L) significantly affected cell viability in both high and low glucose groups , with significant difference between the two groups exposed to 1.0 mmol/L CEES.In high glucose group, 24 h after 0.5 or 1.0 mmol/L CEES exposure, the contents of ATP, ADP and TAN were significantly increased , while ATP/ADP and EC decreased .In low glucose group , ADP, AMP and TAN significantly decreased, while ATP/ADP and EC increased 24 h after 1.0 mmol/L CEES exposure.The mi-tochondrial membrane potential (MMP) also changed differently after 0.5 mmol/L CEES exposure.MMP in high glucose group marginally increased at 3 h, and significantly increased at 8-12 h (P<0.05), and returned to normal at 24 h. MMP in low glucose group showed a transient decrease at 5 h (P<0.01), and back to normal at 8 h.The protein levels of COX-10 and ISCU were significantly increased in high glucose group 24 h after 0.5-1.0 mmol/L CEES exposure , but sig-nificantly decreased in low one 24 h after 1.0 mmol/L CEES exposure .Conclusion When 16HBE is cultured at a high or low glucose concentration , the cell growth, stress responses and energy metabolism including MMP , COX-10, ISCU and ATP production are in different status before or after CEES exposure .High glucose could protect against CEES exposure .