Objective:To stimulate the PBMCs of systemic lupus erythematosus patients with recombinant human high mobility group box1 (HMGB1) protein,observe the effect of HMGB1 on the expression of LC3Ⅱ/Ⅰ protein in active lupus nephritis and inactive lupus nephritis patients.Evaluate the effect of recombinant human high mobility group box 1 ( HMGB1 ) on the proliferation of PBMCs in patients with SLE.Methods:Western blot was used to detect the expression of LC 3II/I protein in PBMCs of active lupus ne-phritis and inactive lupus nephritis patients after stimulated by 1 μg/ml HMGB1 for 0,6,24 and 48 h.CCK-8 assay was used to detect the effects of PBMCs proliferation in patients with SLE after 1 μg/ml HMGB1 stimulation 72 hours.The statistical software SPSS17.0 was used to analyzed the results.Results: Western bolt showed an increasing expression of LC 3Ⅱ/Ⅰ protein in SLE patients after stimulated by HMGB1 ( P<0.05 ) , and this effect was time dependent.Compared with inactive lupus nephritis group , the increasing level of autophagy in active lupus nephritis group was more obviously.1 μg/ml HMGB1 could inhibit the proliferation of PBMCs in patients of SLE significantly ( P<0.001 ).Conclusion: HMGB1 may promote autophagy in SLE especially patients with active lupus nephritis and involved in the pathogenesis of SLE and lupus nephritis.Monoclonal antibodies targeting to HMGB 1 or modulators of mam-malian autophagy may provide new way for the treatment of SLE especially LN.

AIM: To explore the effects of high glucose a t fluctuant concentrations on glucose transport activity and expression of phosp hoinositide 3-kinase (p85) in primary cultured skeletal muscle cells. ME T HODS: Rat skeletal muscle cells were cultured at fluctuant glucose conce ntrations (5, 25mmol/L) for 48h. Then the glucose uptake and the expression of p hosphoinositude 3-kinase were measured. RESULTS: The skeletal mu s cle cells treated with fluctuant high concentration glucose showed the impairmen t of the basal and insulin-induced increase in glucose uptake and significant de crease in p85 protein expression as well as p85 mRNA. Particularly, there was a significant decrease in p85 protein and mRNA expression in the high fluctuate l evel of glucose concentration. CONCLUSION: The exposure to fluct uant high concentration of glucose inhibits glucose uptake and induces insulin r esistance in skeletal muscle cells.

Hepcidin are small cationic peptides with antibacterial activity expressed mainly in the liver of living organisms, and they play important roles in the host's immune response against microbial invasion and regulation of iron metabolism. Thus, they are considered to be good substitutes for traditional antibiotics. It is a good choice that the antimicrobial peptides are prepared by recombinant DNA expression. In the present study, two hepcidin mature peptide cDNAs from channel catfish (Ictalurus punctatus) (mCH) and tilapia (Oreochromis niloticus) (mTH) were connected by SOE-PCR in order to obtain more recombinant hepcidin with broad antimicrobial spectrum, and EcoR I and Not I sites were added to 5'- and 3'- ends of the fragment, respectively. The recombinant eukaryotic expression vector "pPIC9K-mCH-mTH" was successfully constructed, and transformed into Pichia pastoris GS115. The transformants containing multicopy gene insertion were selected by using different concentrations of G418 and other specific mediums, and identified by PCR for yeast genomic DNA. Expression was induced by adding 1% methanol at 30 degrees C for different times. Tricine-SDS-PAGE analysis demonstrated that the most appropriate expression time was 72 h, at which a high expression yield (77 mg/L) for the target protein was exhibited. The highly purified target protein was obtained from the fermentation supernatant by SP-Sepharose cation exchange chromatography. Bacteriostatic activity assay demonstrated that the fermentation supernatant containing the target protein and purified recombinant target protein had bacteriostatic activities against gram-positive and gram-negative bacterium. The present result provides the important initial value for industrial production of hepcidin antimicrobial peptide.
Animals ; Electrophoresis, Polyacrylamide Gel ; Fish Proteins ; biosynthesis ; genetics ; Fishes ; genetics ; Gram-Negative Bacteria ; Gram-Positive Bacteria ; Hepcidins ; biosynthesis ; genetics ; Pichia ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis

Objective To construct TIZ gene RNA interference plasmid,as well as study the biological effects of down-regulating of TIZ gene on ovarian cancer cells.Methods According to the mRNA sequence of TIZ on GenBank,3 pairs of small interference RNA (siRNA) for TIZ gene expression were designed and introduced into SKOV3 cells by liposome transfection reagent; the real-time quantitative PCR (QRT-PCR) was used to detect the interference efficiency and selected the most efficiency siRNA segment;pGPU6/GFP/Neo carrier was used to construct pGPU6/GFP/Neo-siRNA-TIZ-573 restructured interference plasmid,introduced into SKOV3 ccll (sh-TIZ-573/SKOV3).The cell growth curves were made by methyl thiazolyl tetrazolium (MTT) method.The clonogenicity efficiency was observed by clony formation test.The cell cycles were measured by flow cytometry (FCM).The ability of invasion,metastasis and adhesion of ovarian cancer cells were detected by the matrigel invasion assay,transwell migration assay and adhesion assay,respectively.Results QRT-PCR results showed that ovarian cancer SKOV3 cells which transfected by recombinant plasmids pGPU6/GFP/Neo-siRNA-TIZ-573 could down-express TIZ gene.The cell growth curves,FCM and clony formation tests showed that the growth and proliferation of sh-TIZ-573/SKOV3 cells were significantly speeded up compared with the control cells (P ＜ 0.01).There was no significant difference in cell invasion,migration and adhesion between sh-TIZ-573/SKOV3 cells [(48.5 ± 1.7) ％,(53.6 ±3.4)％,(64.9 ±5.0)％,respectively] and the control cells (P ＞0.05).Conclusion The growth of ovarian cancer cells could be speeded up by down-expressing of TIZ gene,which TIZ gene may be play a biological role as a tumor suppressor gene.

Objective To observe the change of early postoperative cognition in the elderly under low central venous pressure (LCVP) after lobe resection to investigate the safety and feasibility of LCVP in Geriatrics.Methods A total of 45 ASA class Ⅰ ～ Ⅱ old patients (60 ～ 75 y)with elective liver resection were divided into L group and C group.In the LCVP group (L group),CVP was maintained below 5 cmH2O during liver resection until the lobe was done.The patients in the control group (C group) received standard care (The CVP was controlled between 6 cmH2O to 12 cmH2O).To compare the anesthesia recovery after surgery,all patients were tested with a battery of neuropsychologic assessment of cognitive function preoperatively and on the 7th day postoperatively.Results During liver resection,the MAP [(75.8 ±7.9)mmHg] and CVP [(3.1 ±0.4)cmH2O] of experimental group were lower than the control group [MAP (92.3 ± 10.6)mmHg,CVP(9.3 ± 1.4)cmH2O].The difference was statistically significant (t' =20.08,P ＜0.05,t =5.89,P ＜0.01) ;There was no statistically significant difference in postoperative recovery of spontaneous breathing,respiratory,eye opening time,extubation time and leave the operating room time between two groups[(18.1 ±6.7)min,(25.4±8.3)min,(31.9±11.7)min,(42.8±17.8)minvs (15.3 ± 7.0)min,(22.6 ±9.4)min,(30.2 ± 10.8) min,(45.4 ± 13.6) min,P ＞ 0.05].The incidence of POCD was 30.0％ in the experimental group and 27.3％ in the control group.The difference was no statistical significance between two groups(P ＞0.05).Conclusious There was no significant influence of low central venous pressure on anesthesia recovery time and early postoperative cognition in the elderly under hepatic resections.

Objective To analyze the association of staphylococcal nuclease domain-containing protein 1(SND1) and T-cell intracellular antigen 1(TIA-1) on stress granules, and the regulation of SND1 on stress granules under stress stimuli. Methods The immunofluorescence assay and laser scanning confocal microscopy were used to observe the co-localization of SND1 protein and TIA-1 protein under stress stimuli, and the over-expression plasmids of pEGFP vector were transfected into HeLa cells and to verify which domain of SND1 co-localized with TIA-1 under stress stimuli. RNA interference-mediated knockdown of the expression of SND1 protein in HeLa cells was measured by Western Blotting assay. Then whether the knockdown of SND1 affected the recruitment of TIA-1 on stress granules was observed. Heat shocks under different times were used to identify whether there were dynamic changes in transportation of SND1 and TIA-1 on stress granules. Results SND1 co-localized with TIA-1 on stress granules under stress stimuli, and the associated domain of SND1 were SN domain. TIA-1 still can be recruited on stress granules but a large amount of stress granules were reduced even though the expression of SND1 protein was decreased. And the transportation of SND1 on stress granules was laged behind TIA-1 under different-times of heat shocks. Conclusion SND1 protein co-localizes with TIA-1 on stress granules, and which co-regulates the cellular stress response under stress stimuli.

Objective Clinical data of patients who received rotational atherectomy (RA) combined with drug-eluting stent implantation (DES) in TEDA International Cardiovascular Hospital were retrospectively analyzed to evaluate it's safety,short-term and long-term prognosis.Methods A total of 60 patients who underwent RA were consecutively enrolled in the study in TEDA International Cardiovascular Hospital from September 2012 to September 2015.Pre and post procedure coronary angiography and clinical information were collected.Long term outcomes were obtained by outpatient clinical follow-up or telephone interview.We analysed angiographic data.Results Among 60 patients with 65 lesions,RA combined with drug-eluting stent implantation was successful performed in 62(95.3%) lesions and postoperative stenosis degree drop from (90.7±6.2)% to (19.5±6.6)%.4 cases(6.7%)developed complications and were treated accordingly during procedure with satisfactory results.Overall incidence of in-hospital MACCE was 1.7% with one case (1.7%) of myocardial infarction.The mean follow-up time was (23.3±10.6) months.In stent restenosis occurred in 1 case(1.7%) 10 months after operation.Stent thrombosis occurred in 1 case(1.7%) 16 months after operation and myocardial infarction (MI) occurred in 1 case(1.7%) 7 months after operation.One patient died 14 months after operation and another patient died 17 months after operation.Both of them were considered as cardiac death.There was no mortality of other causes recorded.Long-term MACCE was 9.5% and TLR is 5.1%.Conclusions Rotational atherectomy combined with DES implantation in the treatment of severe coronary artery calcification lesions has high success rate, good safety profile and good short and long-term prognosis.

Objective: To observe the relationship between 1h, 2h post-load plasma glucose (PG) elevation and carotid intima-media thickness (CIMT) in healthy population.
Methods: The healthy subjects from normal physical examinations in our hospital from 2013-09 to 2015-05 were selected and oral glucose tolerance test (OGTT) was conducted in them, plasma levels of fasting glucose (FPG), 1hPG and 2hPG were monitored. Finally, 482 subjects with normal fasting glucose, without diabetes were enrolled and divided into 4 groups:①Normal glucose tolerance (NGT) group,n=201,②1hHPG group, the subjects with 1h post-load high and 2h post-load normal glucose,n=83,③2hHPG group, the subjects with 1h post-load normal and 2h post-load high glucose,n=101 and④1hHPG/2hHPG group,n=97. The gender, age, BMI, blood pressure and lipid levels in all subjects were recorded, CIMT was evaluated by Doppler ultrasound; the relationship between 1hHPG, 2hHPG and CIMT were analyzed.
Results:①The gender, age, BMI, systemic blood pressure (SBP), DBP and the levels of TC, TG, LDL-C were similar among 4 groups,P>0.05.②For CIMT at both side, both bifurcations and both internal carotid arteries, 1hHPG/2hHPG group was higher than those in 1hHPG group and 2hHPG group; 1hHPG group was higher than 2hHPG group; CIMT in above 3 groups were all higher than NGT group, allP<0.05.③Multivariate logistic regression analysis presented that increased CIMT (CIMT≥1 mm) was related to 1hHPG (OR=2.630, 95% CI 1.497-2.884), 2hHP (OR=1.799, 95% CI 1.350-2.380) and 1hHPG/2hHPG (OR=3.166, 95% CI 2.566-5.231) respectively.
Conclusion: Abnormal 1hHPG and 2hHPG were related to increased CIMT.

Objective To explore the application of LIS management group in hospital software and hardware management.Methods The objective,responsibilities and management procedure of hospital LIS management group were introduced,and its effects were studied in hospital software and hardware management.Results Hospital LIS management group optimized LIS management procedure,decreased fault rate and shortened the time for fault handling.Conclusion LIS management group promotes hospital digitalization and clinical laboratory department management.

Objective · To evaluate the efficacy and prognostic factors of ifosfamide-cisplatin-etoposide (ICE) chemotherapy as salvage regimen for patients with relapsed/refractory diffuse large B-cell lymphoma (DLBCL).Methods · A retrospective analysis was performed on 84 relapsed/refractory DLBCL patients who were treated with ICE salvage regimen at Ruijin Hospital (Shanghai Jiao Tong University School of Medicine,China) from July 2004 to June 2016.Overall survival (OS) was analyzed by Kaplan-Meier method and multivariate Cox proportional hazards models.Results· Of the 84 patients who were treated with ICE regimen,37 (44.0％) patients had responses,including 26 (31.0％) achieving complete remission.The median number of cycles per patient was 3 (range 1-6 cycles).The 1-year and 2-year OS rates were 49.5％ and 30.0％,respectively.The median OS time was 12.2 months.On univariate analysis,patients with early progression/recurrence (P=0.041) and a high-intermediate/high risk according to the international prognostic index (IPI) (P=0.024) and NCCN-IPI (P=0.002) had poorer outcomes.While improved outcome was found in patients in complete remission after chemotherapy (P=0.000).The multivariate analysis revealed that the intermediate-high/high risk according to NCCN-IPI was an independent risk factor,and remission after chemotherapy was an independent prognostic factor for prolonging survival.Conclusion· The ICE regimen can be used as an effective salvage therapy for patients with relapsed/refractory DLBCL.