ObjectiveTo investigate the expression characteristics of Caspase-8 in gastric carcinoma tissues and its correlation with clinical pathological features.Methods The mRNA and protein of Caspase-8 in gastric carcinoma and its surrounding tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry,respectively.ResultsThe Caspase-8 mRNA in gastric carcinoma tissues was obviously lower than that in its surrounding ones (0.154±0.065 vs 0.394±0.107,t =14.04,P ＜0.01),which was correlated with lymph node metastasis,tumor stage,differentiation grade (P＜0.01).The Caspase-8 protein in gastric carcinoma tissues was also significantly lower than that in its surrounding ones (31.48％ vs 85.19％,x2 =32.04,P ＜ 0.01),which was correlated with tumor stage,differentiation grade (P＜0.05).ConclusionsCaspase-8 is low-expressed in gastric carcinoma,which may play an important role in the occurrence and progression of gastric carcinoma.

Objective To investigate the effect and safety of primary percutaneous coronary intervention (PCI) of acute ST-segment elevation myocardial infarction (STEMI) in elderly patients.Methods 103 consecutive patients with STEMI treated by primary PCI were divided into two groups according to the age:the elderly group [aged≥65 years,with a mean age of (75.7 ±6.2) years(n =49],the non-elderly group [aged＜65 years,with a mean age of (43.0±8.6) years(n =54].Clinical characteristics,complications related to PCI procedure and success rate were analyzed,and major cardiovascular events (MACE) were followed up for(5.7 ± 1.2) months.Results The proportion of female,patients with Killip ≥ Ⅲ,three vessels disease and higher level of serum brain natriuretic peptide were higher in elderly group than in non-elderly group (all P＜0.05).No significant difference was observed between the two groups in success rate and complications of PCI procedure (both P＞0.05).Patients were followed up for (5.7± 1.2) months.The in-hospital and one-month mortalities were higher in elderly group than in non-elderly group [8.2％ (4 cases)vs.0％ (0 case),10.2％(5 cases) vs.0 ％ (0 case),respectively,all P＜0.05].There was no significant difference in six-month mortality and MACE between the two groups.Multivariate logistic regression analysis showed that Killip ≥ Ⅲ was related with the increase of one-month mortality in patients with STEMI undergoing primary PCI,whereas age was not.Conclusions Primary PCI is effective and safe in elderly patients with STEMI.

Objective To investigate the effect of atorvastatin calcium combined with sitagliptin phosphate on β2-microglobulin (β2-MG ) in peripheral blood and serum uric acid ( UA) levels in patients with type 2 diabetes.Methods A total of 78 patients with type 2 diabetes in endocrinology department from Hangzhou Xiasha Hospital were collected and randomly divided into the control group and the experimental group, with 39 cases in each group.The two groups of patients were treated by conventional treatment, life intervention, the control group were treated by metformin hydrochloride sustained-release tablet and sitagliptin phosphate, the experimental group were treated on the basis of control group with atorvastatin calcium.Both groups were treated for 3 cycles, one cycle for 7 days.The clinical curative effect,β2-MG in peripheral blood and urine, serum UA levels and adverse reactions were compared between two groups after treatment.Results After treatment, compared with control group, the clinical total effective rate in experimental group was higher (P<0.05).The serum IgG,β2-MG, urineβ2-MG and UAlb, serum UA levels were lower in two groups post-treatment compared with pre-treatment(P<0.05).Compared with control group, serum IgG, β2-MG, urine β2-MG and UAlb, serum UA levels were lower (P<0.05).There was no significant difference between two groups.Conclusion The atorvastatin calcium combined with sitagliptin phosphate has a significant efficacy in the treatment of patients with type 2 diabetes, it could down-regulate β2-MG in peripheral blood and urine and serum UA levels, improve immunity and prevent patients from cardiovascular complications.

Gluconobacter oxydans is known to oxidize glucose to gluconic acid (GA), and subsequently, to 2-keto-gluconic acid (2KGA) and 5-keto-gluconic acid (5KGA), while 5KGA can be converted to L-(+)-tartaric acid. In order to increase the production of 5KGA, Gluconobacter oxydans HGI-1 that converts GA to 5KGA exclusively was chosen in this study, and effects of carbon sources (lactose, maltose, sucrose, amylum and glucose) and nitrogen sources (yeast extract, fish meal, corn steep liquor, soybean meal and cotton-seed meal) on 5KGA production were investigated. Results of experiment in 500 mL shake-flask show that the highest yield of 5KGA (98.20 g/L) was obtained using 100 g/L glucose as carbon source. 5KGA reached 100.20 g/L, 109.10 g/L, 99.83 g/L with yeast extract, fish meal and corn steep liquor as nitrogen source respectively, among which the optimal nitrogen source was fish meal. The yield of 5KGA by corn steep liquor is slightly lower than that by yeast extract. For the economic reason, corn steep liquor was selected as nitrogen source and scaled up to 5 L stirred-tank fermentor, and the final concentration of 5KGA reached 93.80 g/L, with its maximum volumetric productivity of 3.48 g/(L x h) and average volumetric productivity of 1.56 g/(L x h). The result obtained in this study showed that carbon and nitrogen sourses for large-scale production of 5KGA by Gluconobacter oxydans HGI-1 were glucose and corn steep liquor, respectively, and the available glucose almost completely (85.93%) into 5KGA.
Bioreactors ; Carbon ; chemistry ; Culture Media ; chemistry ; Fermentation ; Gluconates ; metabolism ; Gluconobacter oxydans ; metabolism ; Industrial Microbiology ; Nitrogen ; chemistry

AIM:To investigate the synergistic anti-proliferation effect of aspirin and 5-fluorouracil on the co-lon cancer cells and its mechanism .METHODS: Colon cancer cells were divided into 4 groups: control group , aspirin group, 5-fluorouracil group and aspirin +5-fluorouracil group .Synergistic anti-proliferation effect of aspirin and 5-fluoroura-cil on the colon cancer cells was observed by MTT assay .Apoptosis-inducing effect and mechanism were detected by Hoechst 33258 staining, caspase activity assay and flow cytometry analysis .The mRNA and protein levels of apoptosis-re-lated proteins were evaluated by real-time PCR and Western blotting .RESULTS:5-Fluorouracil inhibited proliferation of HCT116 and SW620 colon cancer cells effectively , and low concentration of aspirin exerted synergistic inhibitory effect .5-Fluorouracil induced apoptotic morphology and increased caspase activity and sub -G1 phase in HCT116 cells.The synergis-tic effect of aspirin obviously enhanced apoptotic ratio and caspase activity .Moreover , 5-fluorouracil inhibited the mRNA and protein expression of Bcl-2, which was amplified by low concentration of aspirin .CONCLUSION:Aspirin and 5-flu-orouracil had a synergistic anti-proliferation effect on the colon cancer cells through apoptosis pathway .

0bjective To establish a standard curve method with more accuracy employed in fluorescence real-time PCR(RT-PCR)as a alternation of the general method.Methods β-actin and KLK11 plasmid DNA for quantitative standard curve were constructed in our study,and Plasmids of β-actin was employed as a internal control.After serial dilution these plasmid were used as DNA standard to obtained slope.Expression of these two genes in malignant prostate cancer cell line LNCAP were tested by real-time PCR,and we analyzed the RT-PCR results with two different methods and compared their accuracy.Results Thestandards curves made from these linear DNA standards showed good linearity (R2=0.991 and 0.992 for β-actin and KLK11 standards graphs),but also displayed a discrepancy in their PCR efficiency(β-actin 123% and KLK11 99%).There were different results after two different stand curve analytical method:the expression of KLK11 mRNA in LNCAP was downregulated in general standard curve method.In the new analytical method,howerer,KLK11 upregnlated for 4.46-fold.And there was a significant difference between aplification efficiency of targt gene and internal control gene(t=4.829,P<0.05).Conclusions Compared with general standard curve method,the new advanced standard curve method described here avoids an error which considers there is identical amplification efficiency between target gene and internal reference gene.It is considered to be a more correct analytical method in fluorescence real-time PCR.

ObejectiveTo evaluate the efficacyofacupointthread embedding in easing painafterMilligan-Morgan(M-M)for mixed hemorrhoids.MethodSixty patients undergone M-M for mixed hemorrhoids were randomized into a treatment group of 30 cases and a control group of 30 cases. After M-M, patients in the treatment group received thread embedding at Changqiang (GV1) and bilateral Zhibian (BL54), while the control groupdidn’treceive any intervention. The onset time of post-operative pain, average pain index within a week, and pain index after defecation, electromyogram (EMG), change of anal canal pressure, patients’ satisfaction, and adverse-event rate were observed.ResultThe average pain index and pain index after defecation in the treatment group were significantly lower than that in the control group (P<0.05), and the onset of pain in the treatment group was significantly later than that in the control group (P<0.01); after surgery, the anal canal resting pressure in the treatment group was markedly lower than that in the control group (P<0.05); there wasno significant difference in comparing the squeeze pressure of anal canal between the two groups (P>0.05). According to the motor unit potential (MUP) analysis, there were significant differences in comparing the amplitude (Ampl) and Ar/Am of the restingphase between the two groups (P<0.05), while there were no significant differences in comparingthe Ampl, Area, Ar/Am, and Freq of the contraction phase between the two groups (P>0.05). There were significant differences in comparing the patients’satisfaction, adverse-event rate, and use of analgesics between the two groups (P<0.05). ConclusionAcupoint thread embedding can produce a content analgesic effect, and it’s safe and reliable.

Objective To retrospectively assess the prognostic impact of diabetes mellitus (DM) on patients with hepatocellular carcinoma (HCC) after curative resection.Methods A total of 417 HCC patients who had undergone curative hepatic resection were included into two groups.108 patients were classified into DM group and 309 patients into the non-DM group.Overall survival,disease-free survival,postoperative morbidity and mortality were compared between the two groups after reducing confounding bias by propensity score matching.Independent prognostic predictors were determined by Cox proportional hazards model.Results Propensity score matching resulted in 89 patients in each group,and variables were balanced between two groups.In the matched cohort,DM and non-DM groups showed similar morbidity and 30-and 90-day mortality after curative hepatectomy (respectively x2 =0.837,x2 =Fisher,x2 =Fisher,all P ＞ 0.05),the 1-,3-,and 5-year overall survival rates were 82.0％,59.9％,and 33.4％ in DM group and 90.7％,79.1％,and 69.3％ in non-DM group,respectively(P =0.001),however,there was no significant difference in disease-free survival between DM and non-DM groups.Cox multivariate analysis revealed that DM is an independent risk factor for overall survival in patients with HCC after curative resection,but not for disease-free survival.Conclusions DM does not increase the postoperative morbidity or mortality for patients with HCC after curative resection,however,DM may increase the risk of mortality of HCC patients in the long-term.

Objective Our previous study demonstrated that toll-like receptor 2(TLR2) can distinguish extraneous antigen and prevent the immunological response. This study was designed to detect the expression of toll-like receptor 2 (TLR2) mRNA in cornea and investigate the effect of steroid on TLR2 expression in rats cornea following allograft penetrating keratoplasty. Methods The penetrating keratoplasty models were established in SPF rats with the 108 SD rats as receiptors and 36 SPF Wistar rats as donors, and other 6 SPF SD rats worked as normal controls. The receiptor rats were divided randomly into autograft group, allograft group and steriod group. The clarity and neovascularization of corneas of experiment rats were examined under the slit-lamp microscope and the rejection index was calculated based on Holland criteria. Corneal histopathological examination was carried out by hemotoxylin and eosin staining under the light microscope, and real time-PCR was employed for the detect of TLR2 mRNA in the corneas at the fifth, seventh and ninth day after operation. The experimental animals were obtained from the Animal Experimental Center of Southern Medical University and the procedure followed the Statement of Association for Research in Vision and Ophthalmology. Results The rejection occurred in 7 days after operation in allograft group, and only mild edema, opacity and neovascularization of corneas were found at different degrees in 9 days after operation in autograft group and steriod group. Severe corneal edema, a lots of inflammatory cells infiltration and new vessels in stroma were seen in allograft group, and mild inflammatory response was found in autograft group and steriod group. Normal comeal structure was exhibited in normal control group under the light microscope. The fold differences of TLR2 mRNA expression in cornea after amplification was significantly different among three groups and different time points (F_(group) = 39. 46, P = 0. 00; F_(time) =35. 38, P = 0. 00 ; F_(interaction) = 45. 66, P =0. 00), and the evident enhance of TLR2 mRNA expression was revealed in allograft group compared with autograft group (P ＜ 0. 05) and declined in steriod group (P ＜ 0. 05). Conclusion Steriod may restrain the acute allograft rejection by down-regulating the expression of TLR2 in corneas and its signals transaction. This result suggests that steriod offer a protection from rejection of cornea after penetrating keratoplasty.

Objective To investigate the expression of sialic acid-binding immunoglobulin-like lectin-1 (Siglec-1) in the peripheral blood mononuclear cells (PBMCs) in patients with rheumatoid arthritis (RA),osteoarthritis (OA) and healthy controls and to explore the relationship between Siglec-1 expression and disease activity in RA.Methods Siglec-1 protein and mRNA levels were measured by flow cytometry and real-time quantitative reversetranscription-polymerase chain reaction (qRT-PCR) in 42 RA patients,28 OA patients and 26 healthy controls,respectively.The correlation studies between Siglec-1 and disease activity score 28 (DAS28) or C-reactive protein were performed.T-test was used for comparisons between groups and Pearson's correlation test was used for correlation analysis.Results The percentage of Siglec-1 positive cells of PBMCs in RA group [(15.2±7.6)％] was significantly higher than that in the OA group [(2.3 ±2.6)％] or healthy controls [(2.1±1.6)％,t=8.615,8.661; all P＜0.01].And the major cell type in PBMCs that expressed Siglec-1 was monocytes.The relative Siglec-1 mRNA expression in PBMCs in the RA group (3.4±1.5) was also significantly higher than that in the OA group (1.2±0.4) or healthy controls [(1.0± 0.4),t=3.446,3.966; all P＜0.05].But no significant differences of Siglec-1 protein and mRNA between the OA group and healthy controls were found.Furthermore,positive correlations between Siglec-1 protein and DAS28 or hs-CRP were found in RA patients (r=0.89,P＜0.01; r=0.48,P＜0.01).Conclusion PBMCs are activated which are characterized by elevated expression of Siglec-1 in RA patients.Circulating Siglec-1 may be considered as a potential noninvasive biomarker for monitoring disease activity in RA.