Objective To investigate the effect of splenectomy on peripheral lymphocyte apoptosis and CD4+ CD25+ Treg cells in rat with heart transplantation. Methods Abdominal heterotopic heart transplantation was performed from Wistar (donors) to SD (recipients) rats. Splenectomy was done at the same time in recipients (heart graft splenectomy group), non-splenectomy recipients (heart graft group), single splenectomy in SD rats (spleneetomy group), and SD rats (no operation) were designed as the control group. The transplanted hearts and peripheral blood in each group were collected on the 1st, 3rd, 5th and 7th day after operation. Histopathological and ultrastructural changes in transplanted hearts were observed by microscope and electronic microscope. Apoptosis rate of peripheral lymphocytes and CD4+ CD25+ Treg cells were measured by flow cytometry. The expression of Foxp3 mRNA in CD4+ CD25+ Treg cells was detected by RT-PCR, and survival time of the transplanted hearts was recorded. Results The survival time of the transplanted hearts in heart graft splenectomy group was 17.63±4.54 days, significantly longer than that in heart graft group (7.47±2.24 days) (P<0.05). The transplanted hearts in heart graft group were swelling, hard, dark, with interstitial edema, hemorrhage, diffuse infiltration of inflammatory cells, necrosis and cytolysis of a lot of myocardial cells, blurred cross striations; The transplanted hearts in heart graft splenectomy group were soft, red, partially gray white, with focal edema of subepicardial cells,infiltration of inflammatory cells, intact myocardial cell structure, and clear cross striations; Compared to heart graft group, the ultrastructural changes of transplanted hearts in heart graft splenectomy group were lighter significantly. On the 5th and 7th day after operation, apoptosis of peripheral lymphocytes in heart graft splenectomy group was (7.62±2.15)% and (9.41±3.82)% respectively, significantly higher than in heart graft group (both P<0.05). On the 3rd, 5 th and 7th day after operation, the number of CD4+ CD25+ Treg cells in heart graft splenectomy group was more (all P<0.01), and the expression level of Foxp3 mRNA higher than in heart graft group. Conclusion Splenectomy can increase apoptosis rate of peripheral blood lymphocytes and the number of regulatory T lymphocytes, and up-regulate the Foxp3 mRNA regulation in rats with heart transplantation, which has a negative correlation with pathological changes of transplanted hearts.

Objective To establish a qualitative and quantitative method for controlling the quality of Radix Puerariae and its preparation. Methods The qualitative fingerprint of effective part (extract part of ethyl acetate) was obtained by using RP-HPLC with methanol-water (42∶58) as mobile phase and UV detection at 250 nm . The quantitative measurement of effective components (puerarin and daidzein) was finished under the above chromatographic conditions. And the 11 kinds of different samples of Radix Puerariae were analyzed by the method. Results Under the qualitative conditions, the 5 common peaks in RP-HPLC fingerprint of the effective part could be used as index peaks for qualitative identification. In RP-HPLC quantitative analysis, the recoveries of the method were 98.0% with RSD of 1.30% for puerarin and 95.4% with RSD of 1.61% for daidzein, respectively. The amount of puerarin and daidzein in Radix Puerariae was 0.213%～3.626% and 0.012%～ 0.049% , respectively. Conclusion The analytical method with qualitative fingerprint of effective part and quantitative measurement of effective components of Radix Puerariae can be accurately used for controlling its quality.

Objective To study the application value of atorvastatin combined with anticoagulation drugs in the treatment of steroid-induced necrosis of the femoral head in the elderly.Methods From February 2013 to January 2016, 112 cases elderly patients accepted treatment of steroid-induced necrosis of the femoral head in our hospital were selected.They were divided into observation group and control group with 56 cases in each group randomly.The two groups were treated with anticoagulant therapy, the observation group received atorvastatin on the basis of control group.The efficacy and indicators between two groups after treatment were compared.Results The excellent and good rate was in observation group was 92.86%, which was higher than 66.07% in control group ( P <0.05 ) .The pain degree, walking ability and total joint mobility score of the observation group were significantly higher than those in the control group(P<0.05).After treatment, plasma viscosity and hematocrit of the observation group were significantly lower than the control group(P<0.05).The time of collapse of the femoral head and the index of necrosis of the observation group were significantly lower than the control group(P <0.05).Conclusion Atorvastatin combined with anticoagulant drugs in the treatment of elderly patients with steroid-induced femoral head necrosis has significant effect, could effectively improve the quality of life of patients with good safety, high application value.

Objective To explore the pathogenesis of the superior mesenteric artery syndrome (SMAS) after surgical correction of scoliosis, and to analyse the inducing factors of the SMAS during scoliosis correction. Methods From July 1997 to January 2001, 430 cases of scoliosis were corrected with the 3 dimensional derotation technique(312 cases of the CD instrumentation and 118 cases of the TSRH or CDH instrumentation), 5 cases of which suffered from the SMAS postoperatively. Three of these 5 cases had the SMAS during the heavy Halo-pelvic traction following the anterior spinal release, the other two cases happened after the posterior correction. Results For the 5 patients, the symptoms were relieved after fasting,drainage with nasogastric tube, keeping electrolytic balance, left lateral position, suspense or reduction of the heavy traction, all of them recovered completely after 5-7 days. Conclusion SMAS is apt to occur in severe scoliosis, especially in kyphoscoliosis with trunk collapse. If more spine elongation and higher correction rate are obtained during operation, or the heavy Halo-pelvic traction is necessary after the anterior spinal release, the SMAS is more likely to happen. Early diagnosis and treatment play an important role in the recovery of patients.

To study the in situ intestinal absorption kinetics and compatibility influence of peimine and peiminine in rats, the absorption of peimine and peiminine in small intestine (duodenum, jejunum and ileum) and colon of rats was investigated using in situ single-pass perfusion method and the drug content was measured by HPLC-ELSD. Perfusion rate, pH, concentration of drug, gender and bile duct ligation can significantly affect the absorption of peimine and peiminine, the Ka, and Papp values in the condition of pH 6.8 and pH 7.4 had significant difference (P<0.01), as drug concentration irlcreased, the absorption parameters of peimine and peiminine decreased, Ka and Papp between low concentrations and middle concentrations was significant difference (P<0.01). Verapamil can not affect Ka and Papp of peimine and peiminine which are in the extract (P> 0.05). Bitter almonds and licorice can significantly reduce the absorption of peimine and peiminine with the usual dose (P<0.01), extracted separately and together had no significant difference on Ka and Papp (P> 0.05). Experimental results show that the absorption features of peimine and peiminine are basically the same, both of them could be absorbed at all segments of the intestine in rats and had no special absorption window, and with significant differences between male and female individuals. The absorption of peimine and peiminine complies with the active transport and facilitated diffusion in the general intestinal segments. Bitter almond and licorice can reduce the intestinal absorption rate ofpeimine and peiminine.

ObjectiveTo explore the clinical value of ultrasonography in the detection of median nerve in diabetic peripheral neuropathy (DPN).MethodsMedian nerve was detected in 70 healthy adults (control group) and 43 patients with type 2 diabetes mellitus (diabetes mellitus group) by color Doppler ultrasonography.The type 2 diabetes mellitus patients were divided into two groups according to the duration of the disease:＜ 10 years group (26 cases) and ≥ 10 years group( 17 cases).And they were divided into two groups according to the random plasma glucose (RPG):RPG ＜ 15 mmol/L group(28 cases) and RPG≥ 15mmol/L group ( 15 cases).The nerve cross section area (CSA) was taken at the level of the pisiform.Results The median nerve in diabetes mellitus group had decreased echogenicity and increased color in it.The CSA of the median nerve in diabetes mellitus group was significantly higher than that in control group [ ( 10.3 ± 2.1 )mm2 vs. (8.3:± 1.7) mm2 ] (P ＜ 0.01 ).The C SA of the median nerve in ≥ 10 years group was significantly higher than that in ＜10 years group [ ( 12.3 ± 2.0) mm2 vs.( 10.2 ± 2.6) mm2] (P ＜ 0.01 ).The CSA of the median nerve in RPG ≥ 15 mmol/L group was significantly higher than that in RPG ＜ 15 mmol/L group [ ( 11.7 ± 1.8) mm2 vs.(9.6 ± 2.7 ) mm2 ] (P ＜ 0.01 ).ConclusionColor Doppler ultrasonography examination may reveal morphology of median nerve and is useful for the diagnosis of DPN.

ObjectiveTo test the antifungal activity of berberine (BBR) against Candida albicans in vitro. MethodsAccording to the National Committee for Clinical Laboratory Standards (NCCLS) M27-A protocol, broth microdilution test was performed to assess the susceptibility of 30 clinical isolates of Candida albicans to BBR and fluconazole alone or in combination. ResultsThe geometric mean minimum inhibitory concentration (MIC) was 248.18 mg/L and 2.64 mg/L for BBR and fluconazole alone, respectively. When combined with BBR of 320, 160 and 80 mg/L, the geometric mean MIC decreased to 0.03, 0.35 and 1.35 mg/L respectively for fluconazole. Fluconazole and BBR showed synergistic effects in 26.7％ of these isolates including 3 fluconazoleresistant strains, additive effects in 56.7％, indifferent effects in 16.7％, antagonistic effects in none. Conclu sionIn vitro, BBR has antifungal activity against Candida albicans and can enhance the activity of fluconazole against Candida albicans.

Objective To assess the efficacy of emergency hand-assisted laparoscopic hepatectomy for spontaneous rupture of primary hepatocellular carcinoma.Methods A total of 15 eases who underwent emergency hand-assisted laparoscopic hepatectomy for spontaneous rupture of primary hepatocellular carcinoma were retrospectively reviewed.Results All operations were successful.The postoperative mortality rate was.Thirteen cases were followed-up from 11 months to 56 months.The overall 1-,3-year survival rate was 84.6%(11/13),53.8%(7/13)respectively.Conclusion Emergency hand-assisted laparoscopic hepatectomy is a safe and feasible approach for pontaneous rupture of primary hepatocellular carcinoma in clinically selected patients.If the patients condition permits it,emergency hand-assisted laparoscopic hepatectomy should be the treatment of choice.

Objective To develop an improved enzymatic method for assaying creatinine in serum and urine. Methods Cratinine iminhydrolase catalyzes the conversion of creatinine to N-methylhydantoin and ammonia,the later combines with 2-oxoglutarate and NADPH in the presence of glutamate dehydrogenase.Before beginning the reaction sequence,endogenous ammonia is removed by an “ammonia scavenger system”that involves the same auxiliary reaction.The NADPH and 2-Oxoglutarate consumed are restored through a reaction catalyzed by isocitrate dehydrogenase.The activity of this strictly magnesium-dependent enzyme is completely blocked by magnesium-complexing reagent that acts promptly whenever the reaction has started with creatinine iminohydrolase as startor.The reaction is monitored kinetically by measuring the NADPH decrease via its absorbance at 340nm.Results The method affords a simple,rapid,and sensitive assay with good precision and extended linearity.Test results compare closely with the HPLC precedure.The proposed method is not subject to interference from several metablites or from the drugs used commonly in clinics.Conclusion The method is easily automated and is suitable for routine work in clinical laboratories.

Objectives　To investigate the relationship between homocysteine (Hcy), folic acid, vitamin B12(VitB12),lipids and the carotid artery atherosclerosis(CAAs) and the significance of their levels in predicting cardiac and cerebral vascular events of older patients. Methods　Both sides of the common carotid artery and internal carotid artery were measured by B-mode ultrasound in 74 examinees in A, B, C, and D groups and a control group. With fasting serum, cholesterol, triglyceriade, apolipoprotein AI(Apo AI), apolipoprotein B (Apo B), lipoprotein (a) ［Lp(a)］, creatinine, glucose, folic acid, Vit B12 and Hcy were detected. Results　When carotid artery atherosclerosis became severer, serum Hcy was higher. The levels of the control and A, B, C, D groups were (10.2±3.6) μmol/L, (11.5±4.4) μmol/L, (17.9±4.5) μmol/L, (24.7±10.3) μmol/L, and (41.4±22.3) μmol/L respectively. SNK test showed a significant difference between D and other groups (P>0.05). Folic acid and Vit B12 were negatively correlated with the severity of carotid artery atherosclerosis, but only A and D groups showed statistical significance. Spearman correlation coefficient showed that serum Hcy had significant negative correlation with folic acid and Apo AI(P=0.035 and P=0.000).Multiple linear regression analysis between CAAs and various risk factors showed Apo AI had negative correlation, but Apo B, Lp(a) and Hcy had positive correlation with CAAs, especially Hcy(P=0.009).Conclusion　Hyperhomocysteinaemia is an important risk factor of artery atherosclerosis and the degree of CAAs is highly correlated with Hcy concentration. Hcy, Apo AⅠ,Apo B, Lp(a) are sensitive and reliable indexes for evaluating and predicting CAAs.