Objective To explore the effect of c-Jun N-terminal kinase(JNK)on the expression of interleukin-10(IL-10) in keratinocytes induced by ultraviolet A(UVA).Methods The HaCaT cells in cultured were either sham irradiated(control) or exposured to 2.4 J/cm2 UVA radiation.The cells were collected at 0-48 h after irradiation,and JNK levels in cells were detected with the immunofluorescence.HaCaT cells were treated with SP600125(a JNK inhibitor) before irradiation,then cells and suspended medium were collected at each time-point after irradiation,and the expression of IL-10 mRNA and protein were determined by RT-PCR and ELISA.Results Compared with control cells,irradiated cells had increased levels of phospho-JNK throughout the entire 48 h following irradiation(P

Aim To study the effects of luteolin on the expression of TGF-?1 mRNA in rats with pulmonary fibrosis, and elaborate the molecular mechanism of luteolin in pulmonary fibrosis therapy. Methods The model of pulmonary fibrosis was established through instilling bleomycin intratracheally. After luteolin treatment, the pulmonary index and the content of hydroxyproline (HYP) were observed. The levels of TGF-?1 mRNA in pulmonary tissues were determined with RT-PCR. Results The pulmonary index and the content of HYP decreased markedly in pulmonary fibrosis rats treated with luteolin. Furthermore, luteolin inhibited evidently the expression of TGF-?1 mRNA in lung tissues with pulmonary fibrosis. Conclusion The molecular mechanism of luteolin in pulmonary fibrosis therapy was associated with inhibition of the expression of TGF-?1 mRNA.

Object To study the effects of gypenosides on MAO and Na/K ATPase activities in brain tissues of aging mice Methods Models of aging mice were prepared by continuous post orbitol injection of 120 mg/kg D galactose for a month, then the effects of gypenosides on MAO and Na/K ATPase activities in brain tissues were investigated Results It was observed that MAO activity was increased and Na/K ATPase activity was decreased in brain tissues of aging mice models significantly; and these effects were reversible respectively by gypenosides ig of 75 and 150 mg/kg Conclusion The result that gypenosides could invert the changes of MAO and Na/K ATPase activities in brain tissues of aging mice, may provide a further explaination of the anti aging mechanism of gypenosides in molecule enzymology

The main commercial production of fructooligosaccharides (FOS) comes from enzymatic transformation using sucrose as substrate by microbial enzyme fructosyltransferase. A fructosyltransferase genomic DNA was isolated from Aspergillus niger QU10 by PCR. The nucleotide sequence showed a 1 941 bp size, and has been submitted to GenBank (KF699529). The cDNA of the fructosyltransferase, containing an open reading frame of 1 887 bp, was further cloned by RT-PCR. The fructosyltransferase gene from Aspergillus niger was functionally expressed both in Escherichia coli and Pichia pastoris GS 115. The highest activity value for the construction with the α-factor signal peptide reached 431 U/mL after 3 days of incubation. The recombinant enzyme is extensively glycosylated, and the active form is probably represented by a homodimer with an apparent molecular mass of 200 kDa as judged from mobility in seminative PAGE gels. The extracellular recombinant enzyme converted sucrose mostly to FOS, mainly 1-kestose and nystose, liberating glucose. FOS reached a maximal value and represented about 58% of total sugars present in the reaction mixture after 4 h reaction. The results suggest that the availability of recombinant Pichia pastoris as a new source of a FOS-producing enzyme might result of biotechnology interest for industrial application.
Aspergillus niger ; enzymology ; genetics ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Fungal Proteins ; genetics ; metabolism ; Glycosylation ; Hexosyltransferases ; genetics ; metabolism ; Molecular Sequence Data ; Molecular Weight ; Pichia ; Sucrose ; metabolism ; Trisaccharides ; metabolism

Objective The aim of this study was to establish a rat model of blood hypercoagulable state by intra?venous injection of thrombin and to provide a model for researches on hypercoagulable state. Methods Rats were divided into six groups and were injected with normal saline and 2?5, 5, 10, 20, 40 U/kg thrombin solution through the femoral vein, respectively. Then, blood was drawn to test the activated partial thromboplastin time (APTT), prothrombin time ( PT) and fibrinogen ( FIB) , and to observe the death rate of rats in these groups to verify the optimal dosage. On this ba?sis, rats were injected thrombin of the best dose through the femoral vein, and blood samples were collected at 0, 10, 30, 60, 120, 180, 300 (s) to test APTT and PT and FIB for determining the best time for blood sampling. At last, the rats were divided into control group and thrombin group to inject normal saline or thrombin solution in the best dose via the fem?oral vein, and blood was taken at the best time to test APTT, PT, FIB and whole blood viscosity. Results APTT and PT values of the 10 U/kg thrombin group were the shortest, and FIB value of this group was the highest among these groups. APTT and PT values of blood sample collected at about 60 s after thrombin injection were the shortest, and FIB value was the highest. Compared with the control group, PT and APTT values of the thrombin group were shorter (P<0?05), and blood viscosity and FIB were higher ( P<0?05 ) . Conclusions Injecting thrombin solution into the femoral vein can be used to establish a rat model of hypercoagulable state. The best dose of thrombin solution is 10 U/kg in a concentration of 2 U/mL. The best time to collect blood sample is 60 s.

Objective To evaluate postoperative complications of laparoscopy-assisted D2 radical total gastrectomy for gastric cancer as compared with open procedures. Methods In this study,358 patients of gastric cancer undergoing laparoscopy-assisted D2 total gastrectomy or open D2 total gastrectomy between January 2011 and December 2012 were retrospectively reviewed and analyzed.Patients were non-randomly divided into laparoscopic group ( LAP,n =165 ) and open surgery group ( OPEN,n =193 ).Operative time,intraoperative blood loss,postoperative hospital stay and complications were compared between the two groups. Results Operative time [ 225 ( 195 - 340 ) min vs.230 ( 195 - 300 ) min,P ＞0.05 ] and number of lymph nodes dissected [ (26 ± 4) vs.(27 ± 4 ),P ＞ 0.05] between the LAP group and the OPEN group were not significantly different.Compared with OPEN group,blood loss in the LAP group was less [ 160 ( 80 - 600 ) ml vs.270 ( 150 - 600) ml,P ＜ 0.01 ] and postoperative hospital stay was shorter [ ( 11.4 ± 2.6) d vs.( 13.7 ± 2.4) d,P ＜ 0.01 ].Postoperative complications developed in 18 patients in the LAP group and in 30 patients in the OPEN group,the difference was not statistically significant between the two groups. Conclusions In the hands of experienced surgeons laparoscopy-assisted D2 radical total gastrectomy for gastric cancer is safe,effective and miniinvasive.

Objective To determine the effects of pudendal ncrve stimulation with different frequency(5 Hz,20 Hz) on bladder dysfunction dogs after spinal cord injury.Methods The study was done in 4 dogs which chronic spinal cord transection at T9-T10 level.All the dogs underwent electrical stimulation of pudendal nerve (5 Hz,20 Hz).Then,bladder capacity,compliance,non-voiding contraction number (NVC) and voiding efficiency before and during stimulation were compared.Results The bladder capacity and the compliance was increased ( 58.9 ± 17.4) ％ and (53.1 ± 4.9 ) ％ ( P ＜ 0.05 ) by pudendal nerve stimulation with low frequency (5 Hz).Detrusor overactivity can be inhibited and the NVC decreased from 1.7 ± 1.3 to 0.9 ± 1.1.Pudendal nerve stimulation with medium frequency ( 20 Hz) can induce bladder contraction and increased voiding efficiency from (5.8 ± 1.0) ％ to ( 16.3 ± 2.6 ) ％ ( P ＜0.05).Conclusions Pudendal nerve stimulation with low frequency can inhibit detrusor overactivity and increase the bladder capacity and compliance in spinal cord injury dogs.Pudendal nerve stimulation with medium frequency can induce bladder contraction and increased voiding efficicncy.

ObjectiveTo investigate the clinical significance of admission hemoglobin (Hb) value for posttransplantation early respiratory complications in cirrhotic hepatocellular carcinoma (HCC) patients undergoing liver transplantation (LT).MethodsBetween April 2001 and February 2010,the medical record of 100 consecutive liver recipients at First Affiliated Hospital of Guangxi Medial University were retrospectively reviewed. Pulmonary complications developed in 45 patients after LT. Using bivariate correlation analysis between the admission Hb value and pulmonary complications screened for the threshold value of admission Hb value affecting early-phase pulmonary complications.According to the threshold value of admission Hb,LT recipients could be divided into two groups. Twenty-seven peri-operative clinical parameters were analyzed in the two groups.ResultsAdmission Hb ≤ 100 g/L was the threshold value affecting postoperative pulmonary complications.The duration of time to initial passage of flatus and the ICU length of stay were significantly prolonged in patients with admission Hb values ≤ 100 g/L,in which poorer arterial blood gas analyses were common. ConclusionsThe admission Hb value of patients with cirrhosisassociated hepatocellular carcinoma affects the early-phase prognosis after LT.

Objective To study the immunosuppressive mechanism of alkaloid sinomenine (SIN) by observing the effects of SIN on the proliferation and intracellular protein expression levels of nuclear factor of activated T cells (NF-AT) and interferon-gamma (IFN-γ) in CD4+ T lymphocytes of human periphery blood. Methods CD4+ T lymphocytes were isolated from PBMC suspensions with immunomagnetic beads and divided into five groups to culture. (1) Negative control group: no medicine was added to cell culture medium; (2) Positive control group: CsA solution (final concentration: 50ng/ml) was added to cell culture media; (3) Low-concentration SIN group (L-SIN): low-concentration SIN solution (final concentration: 10 μmol/L) was added to cell culture media; (4) Middle-concentration SIN group (M-SIN): middle-concentration SIN solution (final concentration: 200 μmol/L) was added to cell culture media; (5) High-concentration SIN group (H-SIN): high-concentration SIN solution (final concentration: 1000 tmol/L) was added to cell culture media. The proliferations of CD4+ T lymphocytes were observed. Western blotting was performed to detect the protein expression levels of NF-AT. FCM was used to determine the levels of IFN-γ. Results Compared with negative control group, the cell proliferation was significantly inhibited in H- and M-SIN groups (P＜0. 01 ). SIN concentration-dependently inhibited the protein expression levels of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood (P＜0.01). The protein expression levels of NF-AT and IFN-γ were lowest in positive control group. There was a close negative correlation between intracellular levels of NF-AT and cell proliferation inhibition ratio in CD4+ T lymphocytes of human periphery blood (rs = - 0. 969, P = 0. 000). Conclusion SIN can inhibit the protein expression of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood probably by decreasing protein levels of NF-AT to inhibit the activity and proliferation of CD4+ T lymphocytes.

Objective To investigate the short-term therapeutic effect of warfarin in preventing portal vein thrombosis (PVT) after modified laparoscopic splenectomy combined with pericardial devascularization.Methods The retrospective cohort study was used to analyze the clinical data of 32 patients with cirrhotic portal hypertension who were admitted to the Clinical Medical College of Yangzhou University between January 2014 and August 2014.The characteristics of warfarin and aspirin regimens were introduced to the patients before operation for choosing postoperative therapeutic regimen.Based on the decisions, 17 and 15 patients receiving warfarin regimen and aspirin regimen were divided into the warfarin group and the aspirin group, respectively.All the patients underwent successful modified laparoscopic splenectomy and pericardial devascularization with intraoperative autologous blood salvage.The treatments were as follows : from postoperative day 3, patients in the warfarin group received 2.5 mg of oral warfarin once daily with titration of the dose to maintain a target international normalized ratio (INR) of 2.0-3.0 for 1 year;patients in the aspirin group received 100 mg aspirin enteric coated tablets for 1 year;and both groups received 50 mg of oral dipyridamole three times daily for 3 months and subcutaneous injection of 4 100 U of low-molecular-weight heparin (LMWH) once daily for 5 days.Blood cell analysis, liver function, coagulation function and Doppler ultrasound screening for the occurrence of PVT were performed at the first and third months.Postoperative electronic gastroscopy was performed at 3 months postoperatively for observing the change of the esophageal and gastric-fundus varices.The patients were followed up till February 2015.The incidences of PVT and the level of INR at the first week, the first month and the third month after operation were observed.Measurement data with normal distribution were presented as (x) ± s and analyzed by t test, and measurement data with skewed distribution were presented as M(range) and analyzed by the rank-sum test.Comparison of the mean INR at different time points between the 2 groups was analyzed by the repeated measures ANOVA.Comparison of count data was analyzed by the Fisher's Exact Probility.Results There were no gastrointestinal hemorrhage or perioperative death in the 2 groups.(1) The overall incidences of PVT at postoperative week 1 were 9/17 and 6/15 in the warfarin and the aspirin groups, respectively, with no significant difference (P ＞ 0.05).However, the overall incidences of PVT at postoperative month 1 and 3 were 7/17 and 3/17 in the warfarin group, which was significantly different from 12/15 and 12/15 in the aspirin group (P ＜ 0.05).(2)The incidences of main portal vein thrombosis (MPVT) at postoperative week 1 and postoperative month 1 were 5/17 and 6/17 in the warfarin group, 4/15 and 5/15 in the aspirin group, showing no significant difference (P ＞ 0.05).The incidence of MPVT at postoperative month 3 was 3/17 in the warfarin group, which was significantly different from 9/15 in the aspirin group (P ＜ 0.05).(3) The INR was changed from 1.30 ± 0.17 before operation to 1.55 ± 0.38 at postoperative month 3 in the warfarin group, and from 1.33 ±0.14 before operation to 1.21 ±0.11 at postoperative month 3 in the aspirin group, showing significant difference in the changing trend between the 2 groups (F =713.908, P ＜ 0.05).(4) All the 32 patients were followed up for a median time of 7 months (range, 3-11 months).The results of electronic gastroscopy at postoperative month 3 showed that the esophageal and gastric-fundus varices were obviously improved or disappeared.Conclusion Warfarin in preventing PVT after modified laparoscopic splenectomy combined with pericardial devascularization is safe and feasible, with a good short-term outcome.