Ohjective To know the proper wraping method for postoperative patients with neurosurgical intervention treatment. Methods Divided 94 patients with neurosurgical intervention treatment into the ban-dage group and the gauze group randomly, there were 47 cases in the each group. Bandage and gauze was used in the two groups for punctured wound respectively, and then compared the condition of complications and puncture location between the two groups. Results The incidence rate of puncture-associated comphcations in the bandage group was sinificant lower than that of in the gauze group. Conclusions Wraping by bandage for postoperative patients with neurosurgical intervention treatment is more effective than gauze method.

Objective To develop a method for the separation and preparation panaxynol. Methods Panax Notoginseng was extracted by SEF-CO_2, and absorbed by silica gel to remove most of the polar impurities. Panaxynol was preparated with RP-HPLC. Result Panaxynol was identified by UV、IR、MS、NMR,and the purity of panaxynol reached 98%. Conclusion The developed method is simple, reproducible and easy to operate. The solvent is cheap,with low boiling point and easy to recovery property.

Objective To investigate the experiences of combined liver-kidney transplantation (CLKTx).Methods Six patients underwent CLKTx in our center. The primary diseases included chronic glomerulonephritis and post-hepatitis B cirrhosis in 4 patients, hepatitis B virus associated nephritis and primary hepatocellular carcinoma in 1 patient, and polycystic kidney and polycystic liver in 1 patient. Cyclosporine A (or tarcrolimus), mycophenolate mofetil and methylprednisolone were applied to prevent rejection. Four cases of post-hepatitis B cirrhosis received lamivudine. And hepatitis B immunogloblin was given to 4 patients for a short term.Results All 6 liver grafts had good primary function. Five renal grafts had good primary function within one week post-transplantation. One patient with delayed kidney graft function needed supportive hemodialysis. The serum creatinine was declined to normal level 52 days post-operation. Pleural effusion occurred in all 6 patients among which 2 patients needed surgical drainage. Two patients had to be treated for bacterial pneumonia and pneumocystis carinii pneumonia respectively. Three patients needed lipid-lowering therapy at early time post-operation. At the last follow-up, all 6 patients were alive with normal renal graft function and liver graft function. The panel reactive antibody (PRA) of one patient was 23 % before transplantation, and remained at about 8 % post-transplantation. The serum HBsAg and HBV DNA of all 4 post- hepatitis B cirrhosis patients became negative post-transplantation.Conclusion CLTx is a safe procedure for combined hepatic and renal end-stage disease with excellent short-term results.

BACKGROUND: Bone marrow mesenchymal stem cells (MSCs) have been widely accepted by medical investigators due to their advantages including easy obtaining, minimal invasion, with infinite proliferation and multi-differential potential, and without immunological rejection in the autologous transplantation. OBJECTIVE: The goal of this study is to isolate and purify rat bone marrow MSCs in vitro, so as to observe the effects of different concentrations of serum containing kidney-tonifying and blood-activating Chinese herbs on the in vitro proliferation of rat bone marrow MSCs.DESIGN: A randomized controlled animal experiment.SETTING: Hip Center, First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine.MATERIALS: Forty healthy male SD rats of SPF grade, weighing 170 to 180 g, were provided by the Laboratory Animal Center, Guangzhou University of Traditional Chinese Medicine. The protocol was performed in accordance with ethics guidelines for the use and care of animals. The involved rats were divided into 4 groups by random digit table with 10 rats in each: normal control group, high-, middle-, and low-concentration groups. METHODS: This study was performed at the Laboratory Animal Center, Guangzhou University of Traditional Chinese Medicine between January and March 2005. Rat bone marrow MSCs were isolated and purified by Percoll density gradient centrifugation, and cultured in vitro to establish rat bone marrow MSCs culture system. Rats in the high-, middle-, and low-concentration groups were intragastrically administrated with 4.4, 2.2 and 1.1 g/kg serum containing kidney-tonifying and blood-activating Chinese herbs, which equaled to 20, 10 and 5 times of adult dosage, respectively. Rats in the normal control group were intragastrically administrated with purified water for 1 week. One hour after the last administration, 6 mL blood was taken from abdominal aorta of each rat under the aseptic condition. Then, it was centrifuged at 2 000 r/min for 15 minutes, and meanwhile serum was collected. 10% rat serum containing kidney-tonifying and blood-activating Chinese herbs was added to the medium in the high-, middle-, and low-concentration groups, while 10% fetal bovine serum was added in the normal control group. MAIN OUTCOME MEASURES: ① MSCs growth status; ② MSCs morphology was observed by HE staining and Giemsa's staining; ③ MSCs antigen expression was detected by an immunocytochemical method; ④ Effects of different concentrations of serum containing kidney-tonifying and blood-activating Chinese herbs on MSCs growth.RESULTS: ①The primarily cultured bone marrow MSCs began to adhere to the wall 24 hours later and 80% of them reached the confluence 7 days later. ② MSCs took appearance in long shuttle shape or polygon. These cells were little. Nuclei were located in the middle part of cells or a little deviation. The ratio of nucleus to cytoplasm was a little high. ③CD44 expression was found in the cytoplasm of mononuclear cells, and colored blue. Partial MSCs expressed c-Kit. Their cytomorphology and phenotypic expression have the characteristics of MSCs. ④Three days after serum containing kidney-tonifying and blood-activating Chinese herbal medicine being added to high-, middle-, and low-concentration groups, the number of bone marrow MSCs was dose-dependently increased as compared with that in the normal control group. CONCLUSION: Serum containing kidney-tonifying and blood-activating Chinese herbs promotes the in vitro proliferation of bone marrow MSCs.

ObjectiveTo optimize Chonglian oral solution extracting craft. MethodsWith the obtaining rate of extract and the total content of the Pariphyllin Ⅰ and Phillyrin presented in the extract as the indexes for the water extraction process,and with the total content of the Pariphyllin Ⅰ and Phillyrin presented in the extract as the indexes for the alcohol deposition process,orthogonal design was used to optimize the conditions for the extraction process respectively. ResultsThe optimal conditions for the water extraction of Chonglian oral solution was as following:to add water 10 times,decocting 3 times,1 hour each time.The optimal conditions for the alcohol deposition of Chonglian oral solution was as following:concentrated for the relative density to 1.10(80C hot test),cold,add ethanol to the solution for the ethanol content of the solution reached 80%,static settlement for 24 hours. ConclusionThe extracting method is reasonable,stable,and suitable to industrialized producting.

Objective To investigates the gene expression of telomerase by real-time quantitative telomeric-repeat amplification protocol assay (RTQ-TRAP) in the peripheral blood mononuclear cells (PBMC) of colorectal carcinoma patients and the relationship between the telomerase gene expression in PBMC and clinicopathological features. Methods Peripheral blood samptes were collected from 71 colorectal carcinoma patients, 20 benign colorectal disease patients and 25 normal controls. The telomerase gene expression in PBMC was measured by RTQ-TRAP, and serum CEA in colorectal carcinoma patients was measured by chemiluminescence immunoassay. Results The gene expression of telomerase in PBMC was positive in 50 out of 71 cancer patients (70. 4% ) , 1 out of benign patients (5. 0% ), respectively. The difference of the telomerase gene expression of PBMC in benign colorectal diseases and cancer patients was significant (χ2 = 24. 521, P < 0. 001 ). There was no significant association between the expression of telomerase and patient's gender, age, Dukes stage, and tumor site. The positive rate of CEA in colorectal carcinoma patients was not significantly higher than positive rate of telomerase gene expression(χ2 = 2. 286,P = 0. 125). Conclusions The RTQ-TRAP method is highly accurate and sensitive in measuring telomerase gene expression. The detection of telomerase gene expression in PBMC of colorectal carcinoma patients is a simple and useful molecular marker for the diagnosis of colorectal carcinoma.

Objective To evaluate the effects of tadalafil on chronic allograft vasculopathy. Method The abdominal aorta transplantation was performed on Male Lewis or Brown-Norwai rats as donors and male Male Lewis rats as recipients. The recipients were divided into 3 groups: Group A (Lewis-Lewis)with no treatment ;Group B (BN-Lewis) with no treatment; Group C (BN-Lewis) received tadalafil treatment (0.5 mg/kg per day). The rats were sacrificed at 8 weeks post treatment. The grafted aortas were used for histology and Western blot assay. Plasma cGMP level was detected by ELISA assay. Results The aortas intimal in group C was significantly thinner than that in group B. PKG-Ⅰprotein expression in group C was significantly higher than that in group B. Expression of RhoA in group C was lower than that in group B. Conclusion Tadalafil has positive effect on chronic allograft vasculopathy.

Objective To explore the feasibility and effect of combined piggyback orthotopic liver and kidney transplantation in patient with renal graft function failure complicated with cirrhosis. Methods One patient with renal graft function failure complicated with cirrhosis was resected of renal graft. Daily dose of 50?mg CTX was given 5 days after operation and continued for 3.5 months. After two courses of plasma exchange, PRA was reduced from 66?% to 23?% . Combined kidney and liver transplantation was performed simultaneously using piggyback orthotopic liver transplantation technique,and the kidney graft was placed retroperitoneally in left iliac fossa. PRA was monitored every 30 min after liver reperfusion. Postoperative immunosuppressive therapy consisted of FK506, MMF and steroids. Results The kidney and liver grafts functioned normally after transplantation. PRA was reduced from 23?% to 5?% and maintained at 8?% . HBsAg and HCV were returned to negative. Kidney and liver grafts functioned well during a follow up of 3 months. Conclusion Combined liver and kidney transplantation is an effective rescue for loss of kidney graft complicated with cirrhosis, and liver graft can provide protection towards the kidney graft from the same donor.

Objective To develop the hypothesis ‘saturated or non-saturated cytotoxicity model' and explain the various phenomena of antibody mediated immunoresponses in recipients,including rejection and accommodation.Methods The imitating complement dependent cytotoxicity.The threshold set to identify as saturated or non-saturated cytotoxicity depends on antigen-antibody complex(R)whether or not above lethal number(D)in effective time.Feasibility of the hypothesis was examined through explaining various phenomena mediated by anti-donor antibodies,especially some contradictory phenomena.Results Hyperacute rejection,accelerated rejection and acute rejection could be well explained by saturated cytotoxicity.Accommodation of ABO imcompatible transplantion,de novo antibody induced injury,change of protein profile,and C4d deposition in graft could be well elucidated by the hypothesis.Conclusion The hypothesis saturated or nonsaturated cytotoxicity model' help to interpret and interconnect various phenomena of antibodies mediated immune response,such as rejection and accommodation.

Objective To establish a universal stem loop primer (USLP) based real-time PCR method to scan mature miR profile and quantify it's expression.Methods The common universal stem-loop primer pairs were re-designed; 8 random nucleotides were introduced at 3 ' end for reverse transcription of the mature miR,establishing a miR scanning and quantifying system based on SYBR Green Ⅰ PCR (improved USLP method).10-fold gradient diluted standard miRNA-155 cDNA ( 1 ～ 109 copies/μ1) were utilized to evaluate the sensitivity of this method.The specificity was verified by melting curve assay; the precision was assessed by intra-assay coefficient of variation (ICV) of threshold cycle (Ct value) through 20 repeated detections of the standard miR-155 cDNA (2 × 105,2 × 106,2 × 107 copies/μl) ; cost of the primers and time were evaluated,compared with that of the conventional USLP method.Peripheral blood samples were cultured with phytohaemagglutinin (PHA) for0 h,16 h,24 h,48 h and 72 h,and 87 candidate miR that may be associated with human immunity from PubMed data were scanned and quantified from the cultured T cells.Results The sensitivity of the improved USLP method was 103 copies/μl of standard miR-155 cDNA.Melting curve assay showed a single melting peak at 80 ℃,suggesting the excellent PCR specificity of miR-155.Precision of our method quantifying miR-155 was acceptable (ICV ＜ 2.5％ ).Compared with the traditional stem loop primers,our method saved 75％ cost of primers ( 1 917 bp vs 7 851 bp) and 60％ test time of reverse transcription (85 min vs 205 min).By our method,85 of the 87 miR expression in T cells had no significant difference after the PHA stimulation; the expression of miR-150 (72 h) decreased by 10 times and that of miR-155 (48 h) increased 8 times after culture with PHA (Z =-2.032,P =0.042;Z =- 2.023,P =0.043,respectively ).Conclusions The improved USLP method is fast,precise,sensitive,and cost-effective.It could be used for miR profile scanning and quantifying in T cells.