Objective To evaluate the effects of tacrolimus on the secretion of IL-6 and sIL-2R as well as the expression of IL-6 and sIL-2R mRNA by lymphocytes.Methods Jurkat human lymphoma cells were cultured and treated with tacrolimus of different concentrations.Enzyme linked immunosorbent assay was performed to determine the levels of IL-6 and sIL-2R in the supernatant of Jurkat cells at 48 hours after treatment with tacrolimus of 0,10,102,103 and 104 nmol/L,and real time reverse transcription PCR to measure the expression of IL-6 mRNA and sIL-2R mRNA of Jurkat cells at 48 hours after treatment with tacrolimus of 102 nmol/L.Results Tacrolimus of 102 - 104 nmol/L could suppress the secretion of IL-6 and sIL-2R from Jurkat cells (all P＜ 0.05),with a more marked suppressing effect achieved by the use of tacrolimus at 103 - 104 nmol/L.The expressions of IL-6 and sIL-2R mRNA from Jurkat cells were downregulated by tacrolimus of 102 nmol/L (both P ＜ 0.05).Conclusion Tacrolimus at certain concentrations could downregulate the secretion of IL-6 and sIL-2R as well as the expression of IL-6 and sIL-2R mRNA by lymphocytes.

BACKGROUND:Studies have found that both bone marrow mesenchymal stem cels and umbilical cord mesenchymal stem cels have therapeutic effects on liver failure, but little is reported on adipose-derived mesenchymal stem cel transplantation for treatment of liver failure.
OBJECTIVE:To discuss the therapeutic effect of adipose-derived mesenchymal stem cel transplantation on rat liver failure.
METHODS: Forty-five Sprague-Dawley rats were equaly randomized into transplantation, model and control group. Except the control group, animal models of liver failure were made by intraperitoneal injection of D-galactosaminein in the transplantation and model groups. One week after modeling, rats in the transplantation group were given 1 mL of adipose-derived mesenchymal stem celsvia the tail vein.
RESULTS AND CONCLUSION:Adipose-derived mesenchymal stem cell transplantation successfully upregulated alanine aminotransferase and aspartate aminotransferase levels, and moreover, neatly arranged liver cells, reduced inflammatory cells and intact hepatic lobule were shown on B ultrasound after cell transplantation. All these findings indicate that adipose-derived mesenchymal stem cell transplantation has therapeutic effects on rat liver failure by improving rat’s liver function and pathological changes.

The research data at home and abroad show that the overall burnout rate of clinical professional graduate students was high. They are vulnerable to burnout because of long duty hours, learn-ing pressure, intense and overloading work and especially the significantly reduced personal accomplish-ment in the situation of the contradiction between doctors and patients in our country. Burnout not only re-sults in psychological distress and physical symptoms, but also has negative effect on the quality of graduate medical education during residency training. Therefore, educators need to develop an active awareness of burnout and ought to perform interventions such as formulating the appropriate learning goals, improving the work efficiency, reducing work hours, positive psychological counseling and stress management training to prevent such occurrences.

Objective To evaluate the role of C-type lectin domain family 2,member B (CLEC2B) gene in the pathogenesis of vitiligo.Methods Real time fluorescence-based PCR was performed to detect the expression of CLEC2B mRNA in the peripheral blood and lesional skin of 37 patients with vitiligo as well as in the peripheral blood and normal skin of 40 healthy controls.Data were statistically analyzed by t test and chisquare test.Results Among the 37 patients,23 had progressive vitiligo,14 stable vitiligo,31 vitiligo vulgaris,6 segmental vitiligo.The expression level of CLEC2B mRNA was significantly higher in vitiligo lesions than in the control skin (1.21 ± 0.03 vs.1.00,t =4.432,P ＜ 0.05),but was of no significant difference in peripheral blood between the patients and healthy controls (1.02 ± 0.05 vs.1.00,t =1.435,P ＞ 0.05).Increased expression of CLEC2B mRNA was noted in lesions of vulgaris vitiligo compared with those of segmental vitiligo (1.21 ± 0.03 vs.1.02 ± 0.01,t =5.330,P ＜ 0.05),as well as in lesions of progressive vitiligo compared with those of stable vitiligo (1.25 ± 0.05 vs.1.08 ± 0.03,t =3.046,P ＜ 0.05).No significant difference was observed in the expression of CLEC2B mRNA among lesions of vitiligo with different courses (P ＞ 0.05).Conclusion The differential expression of CLEC2B mRNA may take part in the pathogenesis of vitiligo.

OBJECTIVE:To establish a method for the decoction of isoflavone content and its acid hydrolysis conversion rate in the flower of Pueraria lobata. METHODS:Using the flowers of Pueraria lobata as raw material,the isoflavone with main com-ponent of tectoridin in the flower of P. lobata was prepared with ethanol,ethyl acetate extracted,ethanol recrystallized and puri-fied,and it was converted to tectorigenin with hydrolysis in hydrochloric acid. By screening the solvent and wavelength,UV spec-trophotometry was established to determine tectovidin and tectorigenin,and calculate the isoflavone content and acid hydrolysis con-version rate of tectoridin(expressed by the relative percentage of tectorigenin). It was compared with HPLC detection results,the accuracy of UV method was evaluated. RESULTS:The solvent was 70％ethanol solution containing 1％triethylamine,and the iso-flavone content was detected at wavelength of 339,274 nm. The linear range of tectoridin was 8.80-29.33 nmol/mL(r=0.9999). RSDs of precision(n=6),stability(n=5)and reproducibility(n=6)tests were lower than 1.94％;average recovery was 99.7％(RSD=1.77％,n=9). There were no statistical significances in the contents of total flavonoids (UV:17.64-25.55 nmol/mL vs. HPLC:17.39-24.40 nmol/mL) and the relative percentage of tectorigenin (UV:57.65％-87.59％ vs. HPLC:55.62％-91.14％). CONCLUSIONS:The established method is accurate,reliable,and can be used for the rapid determination of acid hydrolysis con-version rate of tectoridin.

Objective To explore an effective method of isolating and culturing the vascular endothelial cells of rat thoracic aorta.MethodThe thoracic aorta was harvested under aseptic condition from the thorax of a Wistar rat.The peripheral connective tissue and fat of the thoracic aorta were stripped and disposed,and then the thoracic aorta was turned inside out to expose the intima.The thoracic aorta was ligated with silk and cauterized on both ends with heated forceps.Then the thoracic aorta was cultured in medium DMEM/F12 containing 20% newborn calf serum in a 50 ml culture bottle which was already blanketed with rat tail collagen.The thoracic aorta was discarded and the new culture medium was added into the culture bottle six days later.The migrating cells were differentially digested by 0.125% pancreatic enzyme for serial subcultivation.The cells were identified by immunohistochemical method with anti-Ⅷ factor antibody.ResultA small amount of cells were seen to migrate from the aorta and adhered to the bottom of culture bottle 6 days after cultivation;the migrating cells spread to cover most part of the bottom of culture bottle 12-14 days later.About 70% of the migrated cells were in a confluent monolayer.The confluent cells grew rapidly after being digested with pancreatic enzyme,and they showed a typical cobblestone appearance.The cells were identified as endothelial cells with 100% expression of Ⅷ factor,which was regarded as the marker of endothelial cells.ConclusionThe method established in the present study is simple and easy to handle,it does not need collagen enzyme and endothelial cell growth promoting substrate,and it is economical and applicable.It is especially suitable for isolation and cultivation of vascular endothelial cells of vessels of small caliber.

Objective To explore the effects of Chinese herbal compound Tangbikang on the expressions of p38 MAPK of sciatic nerve and plasma TNF-α in diabetic rats. Methods Ten of the sixty male SD rats were selected randomly as normal group, and the rest were fed with high-fat diet and low-dosage STZ was used to induce type Ⅱdiabetic rat models. Model rats were randomly divided into model group, mecobalamine group and Tangbikang low-, medium-, and high-dosage groups, 10 rats in each group. Each medication group was intervened with relevant medicine. Rat unilaterals sciatic nerves were taken after 16 weeks. The content of TNF-α in plasma was determined by radioimmunoassay. Western blot method was used to detect the expressions of p38 and p-p38 MAPK protein of sciatic nerve. Results Compared with normal group, the expressions of p38 and p-p38 protein and content of TNF-αin model group significantly increase (P<0.05, P<0.01). Compared with the model group, the expressions of p-p38 protein and the content of TNF-α significantly decreased after medicine intervention in different doses Tangbikang groups and mecobalamin group (P<0.05, P<0.01). The expression of p38 protein in Tangbikang high-dose group significantly decreased (P<0.05), with statistical significance (P<0.05). Conclusion Tangbikang can reduce the expression of p38 and p-p38 MAPK protein of the rat sciatic nerve, and reduce the content of TNF-α protein in rat plasma, which may be one of the effective targets of neuroprotection and abirritation of diabetic peripheral neuropathy.

Objective:To establish an HPLC method for the determination of four kinds of isoflavone compounds including daid-zin, tectoridin, daidzein and tectorigenin in the extracts of Pueraria lobata flowers. Methods:The separation was performed on an Agi-lent C18 column(250 mm × 4. 6 mm, 5 μm) using a mobile phase of methanol-acetonitrile-water(2∶1∶2). The flow rate was 1. 0 ml· min-1 ,and the detection wavelength was 264nm. The column temperature was 25℃ and the sample size was 20 μl. Results:The de-tection could be accomplished within 10 minutes with good separation and specificity of four isoflavone compounds with the retention timeof3.4,3.8,5.7and7.2min,respectively. Thelinearrangewas0.784-78.440 μg·ml-1(daidzin),2.000-200.000 μg· ml-1(tectoridin) and 0.800-80.020 μg·ml-1 (daidzein and tectorigenin),and the relative coefficient was 0.999 9, 0.999 8, 0. 999 7 and 0. 999 9, respectively. The average recovery was 100. 54%(RSD=1. 66%,n=6),100. 03%(RSD=1. 00%, n=6), 99. 48%(RSD=1. 76%, n=6) and 100. 92%(RSD=2. 26%, n=6), respectively. Conclusion: The method is simple, rapid and accurate with good repeatability, which can be used in the rapid determination of isoflavone compounds in the flowers of Pueraria loba-ta.

Objective To compare the influence of hemodialysis (HD) and peritoneal dialysis (PD) on early outcome of patients underwent kidney transplantation from donation after cardiac death (DCD).Methods Patients admitted in the First People's Hospital of Foshan with DCD kidney transplant from January 1st,2011 to June 30th,2016 were analyzed retrospectively.Recipients were grouped into HD group (n=61) and PD group (n=28) according to their pre-transplant dialysis modality.Their short-term outcomes after DCD kidney transplant were compared,including recovery of renal function,short-term complications and laboratory data.Results Patients had longer dialysis duration and lower hemoglobin,serum albumin and phosphorus in PD group than those in HD group (all P ＜ 0.05),but no significant difference shown in age,gender,body mass index,primary disease,blood pressure,and hepatitis B infection (all P ＞ 0.05).HD patients with 6.00(4.00,11.00) d recovery time of renal function,18.00(17.00,21.50) d hospital time,had 24.59％ the delayed graft function (DGF),3.28％ acute rejection and 16.39％ infection during hospitalization.While for PD patients the recovery time of renal function was 4.00(3.75,7.00) d;hospital time was 19.00(15.00,21.75) d;the incidence rate of DGF was 14.29％;acute rejection was 3.57％;and infection during hospitalization reached 17.86％.Above indexes were not significantly different between HD and PD groups (all P ＞0.05).Repeated measure ments showed that,compared with those before transplant surgery,after 1 month,3 months and 6 months HD and PD groups had decreased creatinine and phosphorus,and increased hemoglobinserum albumin and calcium;Serum albumin and calcium were different between the two groups (P ＜ 0.001,P=0.040),whereas creatinine,hemoglobin and phosphorus did not show difference (all P ＜ 0.05).After transplantation the trends of creatinine,hemoglobin,calcium and phosphorus were not different between the two groups (P values were 0.295,0.310,0.501 and 0.063,respectively).Conclusions No significant difference of the recovery regarding renal function,anemia,nutrition status and mineral metabolites was found between pre-transplant HD and PD modality in patients who underwent DCD kidney transplantations.

OBJECTIVETo observe the protective effect of tectorigenin on the vascular endothelial cells(VEC) injured by oxidant low density lipoprotein-cholesterol and the expression of MCP-1 and ICAM-1 mRNA, and explore the mechanism of anti-atherosclerosis.

METHODThe VEC of rat was cultured in vitro and the 100 mg x L(-1) ox-LDL inducing oxidant injured model was used in this study. Different dosage tectorigenin was added into VEC and the activity of VEC was observed by MTT colorimetry. The expression of MCP-1 and ICAM-1 mRNA in VEC was detected by RT-PCR.

RESULTTectorigenin had significantly protective effect on the VEC injured by ox-LDL and obviously inhibited the excessive expression of MCP-1 and ICAM-1 mRNA in VEC.

CONCLUSIONIt was the critical mechanism of anti-atherosclerosis that tectorigenin prevented the VEC oxidant injured and inhibited the excessive expression of MCP-1 and ICAM-1.

Animals ; Atherosclerosis ; drug therapy ; genetics ; metabolism ; Cells, Cultured ; Chemokine CCL2 ; genetics ; metabolism ; Disease Models, Animal ; Endothelial Cells ; drug effects ; metabolism ; Gene Expression ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Isoflavones ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar