Objective To analyze the effect of eukaryotic plasmids containing wild (sense) or anti- sense methylenetetrahydrofolate reductase (MTHFR) gene on cell viability and transcription level of tumor related genes in human gastric cancer cell line.Methods Human gastric cancer cell line MKN-45 was cultured.Recombinant plasmids containing wild MTHFR (W) or antisense MTHFR (A) gene, pCMV-W and pCMV-A,were constructed.Then pCMV-W,pCMV-A and pCMV blank plasmid were transfected into MKN45 cells respectively by using lipofect.Cell viability was analyzed by 3-(4,5-bime- thylthiazolyl-2)-2,5-diphenyhetrazolium dromide(MTT).The transcription levels of Dnmt 1,c-myc, p21~(WAF1) and hMLH1 genes were detected by real-time polymerase chain reaction(PCR).Results Cell vi- ability remarkably increased in those transfected with wild MTHFR (P＜0.01),which was contrary to those transfected with antisense MTHFR(P＜0.01).The expression of those tumor related genes mRNAs were all remarkably decreased in the MKN45-W cells in comparison with those in the MKN45-pCMV cells.No significant difference in the expressions of those tumor related genes mRNAs were found between the MKN45 cells transfected with pCMV-A and blank pCMV.Conclusion MTHFR influences cell viability and the expres- sion level of tumor related genes in human gastric cancer cell line MKN45.

Aim To investigate the expression of mGluR1 in hippocampus of spontaneously epileptic rats(SER) by control study.Methods Total RNA was extracted from hippocampus of SER and Wistar rats,and mGluR1 mRNA expression was detected by RT-PCR;the expression of mGluR1 protein was detected by immunohistochemistry and Western blot.Results mGluR1 mRNA of SER was lower than that of Wistar rats in hippocampus(P0.05) were found.In positive cells,mGluR1 proteins were enhanced.By Western blot,total mGluR1 protein was down-regulated in SER hippocampus(P

Objective To study the epidemiology,pathologies and clinical manifestations of renal diseases in elderly patients of different genders in China northwest region.Methods Retrospective analysis was used to evaluate the clinical and pathological informations of patients above 60 years old who underwent renal biopsy and had the integrated data in the past 20 years.Results (1)Among the 559 patients,there were 347 men accounting for 62.1％,and there were 212 women accounting for 37.9％ ； the average age was (66.7±5.3)years,the median course of disease was 5.0months when receiving renal biopsy,and 50％ cases was (1.3 ～ 12.0)months.(2) Primary glomerulopathy accounted for 69.59％(389/559),secondary glomerulopathy,tubulointerstitial disease and the others accounted for 25.4％ (142/559),3.8％ (21/559) and 1.3％ (7/559),respectively.(3)According to clinical manifestation,nephrotic syndrome was the most common disease(accounting for 60.7％),and then chronic glomerulonephritis accounted for 31.6％.In pathology,membranous nephropathy was the most frequently category accounting for 34.5％,then mesangial proliferative glomerulonephritis accounted for 32.4％.(4) Among secondary glomerulopathy,the frequent diseases were primary polyangitis,diabetic nephropathy,amyloidosis of kidney and lupus nephritis,accounting for 24.7％,19.7 ％,13.4 ％ and 10.6 ％,respectively,and the number of women was more than men in lupus nephritis(P＜0.05).(5) The common diseases of primary glomerulopathy that would result in renal insufficiency were nephrotic syndrome and chronic glomerulonephritis,accounting for 76.5 ％ (26/34)and 76.2 ％ (48/63),respectively,and the common pathological type was both mesangial proliferative glomerulonephritis.The common diseases of secondary glomerulopathy that would result in renal insufficiency were primary small vessel vasculitis and diabetic nephropathy,accounting for 46.3％(19/41)and 34.4％(11/32),respectively.Conclusions (1)the number of men was more than women in this group,and was opposite in lupus nephritis.(2) The most common disease was primary glomerulopathy,as for clinical manifestation,nephrotic syndrome and chronic glomerulonephritis were most common； the most common pathological type were membranous nephropathy and mesangial proliferative glomerulonephritis.(3) The common disease resulting in secondary kidney damage were primary small-vessel vasculitis and diabetic nephropathy.(4)The pathological type of disease resulting in renal insufficiency was mesangial proliferative glomerulonephritis,the common diseases resulting in acute renal insufficiency were nephrotic syndrome,primary small vessel vasculitis,and those resulting in chronic renal insufficiency were chronic glomerulonephritis and diabetic nephropathy.

Objective To assess the disorders of glucose metabolism and insulin resistance in patients with rheumatoid arthritis (RA) and its relationship with disease activity.Methods One hundred and twenty-three RA patients along with 98 age and sex matched controls were studied.Seventy-five g oral glucose tolerance test was performed.The homeostasis model assessment of insulin resistance (HOMA-IR) and beta cell function (HOMA-β) were evaluated.Disease activity score (DAS28) was used to assess disease activity.According to their DAS28 values,patients were divided into high disease activity group and low to moderate disease activity group.Glucose tolerance and HOMA-IR were compared between the two groups.Parameters that reflects disease activity,such as CRP and ESR,as well as disease activity scores were compared between patients with T2DM or prediabetes and patients with normal glucose tolerance.The data was analyzed by t test,Pearson correlation analysis and chi-square test.Results The prevalence of T2DM [20.3％(25/123) vs 5.1％ (5/98),x2=10.774,P＜0.01] and prediabetes [39.0％ (48/123) vs 7.1％ (7/98),x2=29.657,P＜0.01] increased in RA patients compared to controls.RA patients had higher HOMA-IR (2.5±1.5 vs 0.8±0.4; t=5.185,P＜0.01) and lower HOMA-β (83±69 vs 192±85; t=3.768,P＜0.01) compared to controls.ESR [(55±30) mm/1 h vs (37±26) mm/1 h; t=3.159,P＜0.01],CRP [(40±23) mg/L vs (19±10) mg/L; t=3.628,P＜0.01] and DAS28 score (5.6±1.3 vs 4.8±1.2; t=2.923,P＜0.01) were higher in RA patients with T2DM or prediabetes than in RA patients with normal glucose tolerance.In RA patients,the HOMA-IR was significantly positively correlated with DAS28 (r=0.39,P＜0.01),ESR (r=0.54,P＜0.01)and CRP (r=0.20,P＜0.05).The HOMA-IR value and fasting insulin levels were higher in high disease activity patients (DAS28＞ 5.5) than in low-to-moderate disease activity patients (DAS28 ≤5.5) although fasting plasma glucose level did not differ significantly in these two groups.Conclusion The prevalence of T2DM and prediabetes increases in RA patients comparing to controls.RA patients have insulin resistance that is associated with disease activity and systemic inflammation.

Objective To study the characteristics of Genetic typing and the antibiotic susceptibility testing of strains from Pseudomonas aeruginosa keratitis patients.Design Experimental study,Participants 23 eyes of 23 patients of Pseudomonas aeruginosa keratitis.Methods The genomic was extracted and amplified with PCR.The PCR products were purified and sequenced.The results were registered in MIST web Antibiotic susceptibility testing were performed in theses strains,Main Outcome Measures Sequence types and antibiotic susceptibility.Results The isolates were resolved into 20 STs.Two lineages were identified.MIC test showed that strains were more susceptible to aminoglycosides,The activity of quinolones and cephalosporin were higher than that of aminoglycosides.Conclusion MIST can determine homology of the strains from Pseudomonas aeruginosa keratitis by clustering results. There is no finding about relationship between Genetic typing and drug resistance.

Objective To investigate the variation of immune index in patients with systemic lupus erythematosus (SLE) treated with autologous purified CD+34 cells transplantation and to clarify the relationship with pathogenesis and prognosis. Methods Flow cytometry (FCM) and enzyme linked immunosorbent assay(ELISA) were used to test lymphocyte subsets, C3, C4, CH50, autoantibodies and immunoglobulin for 18 cases of SLE before and after transplantation. Results The results showed that the ratio of all the T cell subsets reduced obviously in early postgraft and recovered gradually in 1 to 3 months after transplantation except CD45 RO+CD+4 cells. The levels of serum C3, C4, CH50 increased significantly after transplantation. No case relapsed within one year after transplantation, but 2 patients relapsed one year after transplantation. The levels of the indexes in the patients with relapse were significantly lower than those in the patients with persistent remission, including C4 in the entire course, CH50 in the 3rd and 12th month after transplantation and CD45 RA+ CD+8 cells in the 6th month after transplantation. However, the ratio of CD45 RO+ CD+4 cells in the first month after transplantation in the patients with relapse was higher than that in the patients with persistent remission. Conclusion Autologous purified CD+34 cells transplantation is effective for treating SLE. Survey of immune indexes before and after transplantation is important to investigate the pathogenesis of SLE. Moreover, these immune indexes can be used to predict therapeutic efficacy of SLE.

Objective To select the herba anti- human papillomavirus (HPV) active fraction from herba Arnebia. Methods The fractions from herba Arnebia. were separated with systematic solvents including petroleum arieal part of benzin, n- butanol , ethanol and distilled water, and their effects on HPV- DNA were evaluated by fluorescence quantitative polymerase chain reaction (FQ- PCR) technique. Results Only the water- extract of.this drug showed in- vitro inhibitory effect on HPV- DNA and its minimum effective concentration is 0.08g/mL. Conclusion Herba Arnebia. has in- vitro inhibitory effect on HPV- DNA and the active components exists in the water- extract.

Objective To observe and comparatively analysize the effect of intraperitoneal injection of hydrogen and hydrogen-rich saline in neonatal rats with hyperoxia-induced lung injury.Methods Sprague-Dawley(SD) newborn rats were randomly divided into six groups(n=10).air group(A),air+ hydrogen-rich saline group(B),air+hydrogen group(C),hyperoxia group(D),hyperoxia+hydrogen-rich saline group (E) and hyperoxia+ hydrogen group(F).The group A,B and C were exposed to air and group D,E and F were exposed to 95 ％ oxygen.The group B and E were intraperitoneally injected with hydrogen-rich saline (10 mL/kg,twice daily),while the groups C and F with hydrogen (10 mL/kg,twice daily).The group A and D were injected with normal saline(10 mL/kg,twice daily).Lung tissue and serum samples were collected on 15 d of experiment.The pathological changes of lung tissue and radiate alveoli count (RAC) were observed by HE staining.The content of HYP in lung tissue was detected by the alkaline hydrolysis method,serum SOD and MDA levels were measured.The expression of α-SMA in lung tissue was detected by immunohistochemistry SP method.Results Compared with the A group,RAC and SOD activities in the D group were significantly decreased,while the HYP and MDA levels and α-SMA expression were significantly increased.Hydrogen intervention could significantly alleviate these changes caused by hyperoxia.while intraperitoneal injection of hydrogen got better effect than intraperitoneal injection of hydrogen-rich saline.Conclusion Hydrogen can extenuate the indexes of hyperoxia-induced lung oxidative damage,impairment development and fibrosis to a certain extent.Intraperitoneal injection of hydrogen has slightly better effect than hydrogen-rich saline.

Objective To evaluate the effectiveness and safety of gefitinib retreatment beyond progression(GRBP)in non-small cell lung cancer(NSCLC)patients with rare EGFR mutations.Methods We retrospectively analyzed six rare-EGFR-mutation NSCLC patients from Jan 2011 to Dec 2015.Those patients had previous disease control and then disease progression according to Response Evaluation Criteria in Solid Tumors version 1.1(RECIST v1.1)after taking oral gefitinib 250 mg once a day.After that,continuing gefitinib was decided by clinicians′ experience at the same treatment option.The primary endpoints were response rate(RR),overall survival(OS),the first and second progression-free survival(PFS-1 and PFS-2).Safety was assessed according to the NCI-CTCAE version 4.0.Results After initial treatment of gefitinib,4 patients achieved partial response(PR)and 2 patients showed stable disease(SD),with RR being 66.7％.The median PFS-1 and PFS-2 were 10 months(95％CI 6.6-13.4)and 9 months(95％CI 6.9-11.1),respectively.The median OS time was 28 months(95％CI 10.4-45.6).The most common treatment-related adverse events were fatigue,diarrhea,rash,itching and elevated transaminases.Conclusion In our study,gefitinib retreatment beyond disease progression is effective with a manageable tolerability profile.

OBJECTIVE: To detect the serum level of a novel autoantibody, anti-tubulin-α-1C, in patients with systemic sclerosis (SSc) and to investigate its clinical significance. METHODS: Anti-tubulin-α-1C antibody levels were determined by enzyme-linked immunosorbent assay (ELISA) in 62 patients with SSc, 38 systemic lupus erythematosus (SLE), 24 primary Sjögren's syndrome (pSS) patients, and 30 healthy controls (HCs). Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), immunoglobulin A(IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), C3, C4, rheumatoid factor (RF), antinuclear antibody(ANA), anti-centromere antibodies(ACA), anticardiolipin (aCL), anti-dsDNA antibody, anti-Sm antibody, anti-RNP antibody, anti-Scl-70 antibody, anti-Ro52 antibody, anti-SSA antibody, anti-SSB antibody, centromere protein A(CENP-A), centromere protein B (CENP-B) were measured by standard laboratory techniques. Raynaud's phenomenon and modified Rodnan skin score(MRSS) were recorded to evaluate the disease status of SSc. Independent sample t test, Chi square test, Mann-Whitney U test, Spearman rank correlation were used for statistical analyses. RESULTS: The serum anti-tubulin-α-1C antibody concentration in SSc group was 81.24±34.38, the serum anti-tubulin-α-1C antibody concentration in SLE group was 87.84±38.52, the serum anti-tubulin-α-1C antibody concentration in pSS group was 59.79±25.24, and the serum anti-tubulin-α-1C antibody concentration in healthy group was 39.37±18.7. Multivariate analysis revealed that anti-tubulin-α-1C antibody levels were significantly increased in the SSc and SLE patients. The expression level of anti-tubulin-α-1C antibody in SSc was higher compared with the pSS group and the health control group (P < 0.01). Further analysis demonstrated that the elevated anti-tubulin-α-1C antibody were correlated with the SSc inflammation and disease activity markers ESR(r=0.313, P=0.019), The levels of anti-tubulin-α-1C antibody were also significantly correlated with MRSS(r=0.636, P < 0.01). The best cut-off value for the diagnose of SSc was 76.77 as mean+2SD value. The proportion of Raynaud's phenomenon was higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than that in anti-tubulin-α-1C autoantibody negative group(71.4% vs. 37.5%, P=0.039). The proportions of anti-Scl-70 antibody, anti-CENP antibody and anti-cardiolipin antibody were higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than in the anti-tubulin-α-1C autoantibody negative group (37.9% vs. 15.2%, 34.5% vs. 12.1%, 13.8 vs. 0, respectively, all P < 0.05). CONCLUSION Based on this explorative stu-dy, the level of anti-tubulin-α-1C antibody increased in the serum of the patients with SSc. There were correlations between anti-tubulin-α-1C autoantibody and clinical and laboratory indicators of the SSc patients. It may become a novel biomarker indicative of active SSc and could be applied in future clinical practice.
Antibodies, Antinuclear ; Autoantibodies ; Humans ; Lupus Erythematosus, Systemic ; Scleroderma, Systemic ; Sjogren's Syndrome