Objective:To obtain monoclonal antibodies(McAb) against Staphylococcal enterotoxin C2(SEC2) and establish method for detecting SEC2.Methods:The secreted SEC2 from staphylococcus aureus was used as antigen to immune BALB/c mice. Monoclonal antibodies against SEC2 were prepared by normal hybridoma technique. By identifying the characters of McAbs, the quantitative detection ELISA test method were established and were preliminarily applied.Results:Four hybridmas producing antibodies against SEC2 were obtained. IgG isotypes of four McAbs were IgG1. Their binding site was different except two McAbs that shared the same binding site.The McAbs were proved to be specific for SEC2.The sandwich ELISA method had good specificity, sensitivity and reproducibility, and it was founded to be able to detect SEC2 at concentration from 0.5 to 20 ng/ml. Its recovery ranged between 97.8% and 101%,and CV value ranged between 2% and 5%.Conclusion:The prepared McAb against SEC2 can be used for SEC2 immunoassay. This work provides a basis for controlling the quality of JINPUSU and researching staphylococcal enterotoxin.

Objective:To develop a real-time PCR assays based on TaqMan chemistry for the identification of recombinant baculovirus and determination of virus physical titers in Bac-to-Bac system.Methods:The recombinant baculovirus containing human IL-18 gene was produced using Bac-to-Bac system.A 10-fold series diluted primary viral stocks were used for plaque assay and DNA extraction.Bacmid(baculovirus plasmid) was 10-fold series diluted and served as standards.Real-time PCR amplification of the IL-18 gene was performed in triplicate for each diluted recombinant virus.At the same time,plaque assays were performed using overlay agarose method.Results:The standard linear(101 to 108 copies) from quantitation was achieved with the standard curve.We also find that the "vg/ml" titer value is generally about 10 times than "pfu/ml" titer of the same recombinant virus stock.Conclusion:A TaqMan real-time PCR method is established to identify the recombinant baculovirus and determine the "vg/ml" titer of virus.The method is rapid and quantitative over a wide range of virus titers.

Objective:To express and purify a new fusion protein harboring human epidermal growth factor receptor(EGFR) binding domain and Interleukin-18(IL-18), as well as preliminary assay biological activity of recombinant proteins.Methods:Fusion protein was expressed in insect cells Spodoptera frugiperda cell (Sf9) by using Bac-to-Bac system, and an abbreviation purification procedure was used to purify fusion protein. IFN-?induction assay and EGF Receptor competitive test was used to determine fusion protein's biological activity.Results:SDS-PAGE and western blot assay showed that purified EGF-IL-18 fusion protein had high purity in 20 kD as expected and had the same antigenic specificity as human IL-18. IFN-?induction assay and EGF Receptor competitive test showed that fusion protein induced production of IFN-?in human PBMC and bound with tumor cells.Conclusion:EGF-IL-18 fusion protein has been successfully expressed and purified in insect cells and shows potential to apply in targeting therapy for tumor.

AIM: To study the effect and mechanism of chlorophyllin (CHL) inhibiting HT29 cells. METHODS: IC 50 value and growth curve of HT29 cells were detected with MTT method. Apoptosis was detected with Wright-Giemsa staining, FCM and DNA electrophoresis. Telomerase was detected by PCR-ELISA, and protein and mRNA expression of COX-2 gene were detected through RT-PCR and Western blot. RESULTS: CHL inhibited the growth of HT29 in a dose-dependent manner. CHL blocked HT29 cells in G 1 phase but did not induce apoptosis. Different concentration of CHL inhibits the expression of telomerase and COX-2 in HT29 cells. CONCLUSION: CHL inhibits the growth of HT29 cells by inhibiting the expression of telomerase and COX-2 and blocking cells in G 1 phase. [

Objective To observe the level of reduced glutathione(GSH) and oxidized glutathione(GSSG)in a mouse bone cell line MC3T3-E1 cells exposed to fluoride.Methods MTT method was used to detect cell viability of M C3T3-E1 cells exposed to varying concentrations and periods of fluoride [F-concentration:0(control),0.5,1.0,2.0,4.0,8.0,12.0,20.0 mg/L; F-periods:1,2,4 and 10 days].The Xevo TQ MS was employed to test the levels of GSH,GSSG and glutamine (Gln).Results The MC3T3-E1 cell viability was significantly higher in the 2 mg/L group(0.57 ± 0.05) 1 day after the exposure compared to the respective control(0.49 ± 0.03,P ＜0.01); conversely,cell viability was markedly lower in the 8 mg/L(0.49 ± 0.07) and 12 mg/L(0.47 ± 0.09)groups 4 days after the exposure in comparison to the control(0.63 ± 0.06,P ＜ 0.05 or P ＜ 0.01).The cell viability in the 8 mg/L group(1.52 ± 0.29) 10 days after the exposure was significantly higher than that in the control group (0.86 ± 0.23,P ＜ 0.01),however,the value in the 20.0 mg/L group (0.54 ± 0.07) was significantly lower(P ＜0.01).The level of cell GSH decreased significantly in the 20 mg/L groups 2 days[(13.92 ± 4.63)μmol/L]and 10 days [(0.53 ± 0.30)μmol/L]after exposure compared to the respective comtrols [(26.42 ± 3.67),(24.85 ± 5.68)μmol/L,all P ＜ 0.01].The level of cell GSSG markedly increased in the 2 mg/L group 2 days [(1.12 ± 0.62)μ mol/L]and the 8 mg/L group 4 days [(2.13 ± 0.62)μ mol/L]after exposure compared to the controls[(0.55 ± 0.22),(1.46 ± 0.46)μmol/L,all P ＜ 0.05].The similar change was observed in the 8 mg/L group[(2.97 ± 1.30)μmol/L] 10 days after exposure compared to the control [(1.35 ± 0.50)μmol/L,P ＜ 0.05].The level of Glndecreased significantly in the 2 mg/L group[ (62.80 ± 17.4l)μ mol/L] 4 days and in the 8 and 20 mg/L groups 10 days[ (122.26 ± 19.51), (19.38 ± 8.11)μmol/L] after exposure compared to the controls [ (83.28 ±14.32), ( 147.15± 16.95) μmol/L , all P ＜ 0.05 or P ＜ 0.01 ]. Conclusions Fluoride exposure can significantly promote the changes of GSH, GSSG and Gln levels in the osteoblast, thus affecting the intracellular redox equilibrium.

Background Nd: YAG laser posterior capsulotomy is an important way for after cataract.Usually the patient will use glucocorticoid eye drops to treat the anterior chamber inflammation after operation,but there is potential risk of elevating intraocular pressure (IOP).Objective This study was to compare the clinical effectiveness and safety of loteprednol etabonate ophthalmic suspension,tobramycin+ dexamethasone eye drops and fluorometholone eye drops following Nd: YAG laser posterior capsulotomy.Methods A randomized-controlled clinical trail was performed.One hundrcd and seventy-onc cycs of 127 paticnts who received Nd: YAG laser posterior capsulotomy for after cataract were randomly divided into four groups.Loteprednol etabonate ophthalmic suspension,fluorometholone eye drops,tobramycin+dexamethasone eye drops and systane eye drops was topically administered respectively in the four groups after laser posterior capsulotomy and 6 times per day for 5 days.IOP was measured with Goldmann tomometer 1 hour before operation and 1 hour,1 day,3 days and 7 days after operation.The ocular anterior segment inflammatory response was examined under the slit lamp and scored based on the Peizeng criteria.Written informed consent was obtained from each patient before any relevant medical procedure.Results The IOP was (18.2 ±4.7),(20.1 ±5.7),(18.7±5.5),(19.0 ±4.1),(19.5 ±3.5) mmHg in various time points in the loteprednol etabonate group; (18.7 ±5.3),(20.9±5.7),(21.3±4.5),(21.0±4.9),(22.5±6.5) mmHg in the fluorometholone eye drops group ; (17.9± 6.3),(20.3 ± 6.1),(23.0 ± 3.7),(24.7 ± 4.9),(24.5 ± 6.5) mmHg in the tobramycin +dexamethasone group and(18.4±6.3),(20.7±3.7),(22.7±6.5),(19.6±4.8),(18.5±3.5) mmHg in the systane group,showing a significant difference among the 4 groups (Fgroup =3.876,P =0.023).With the time lapse,the IOP was gradually reduced in the loteprednol etabonate group and systane group,but that in the fluorometholone group and tobramycin+dexamethasone group was elevated,showing a significant difference among them (Ftime =3.801,P =0.031).No any ocular and systemic adverse effect was found in various groups.The percentage of grade 1 and 2 of aqueous inflammatory cells was lower in the loteprednol etabonate group and tobramycin+dexamethasone group than the fluorometholone group and fluorometholone group and systane group(H =8.276,P =0.012).The percentage of Ⅰgrade of aqueous flare was 8％ in the loteprednol etabonate group,22％ in the fluorometholone group,18％ in the tobramycin+dexamethasone group and 30％ in the systane group,with a significant difference among them (H=9.305,P=0.000).Conclusions The use of corticosteroid eye drops can relieve the inflammatory response of ocular anterior chamber after Nd: YAG laser posterior capsulotomy.Loteprednol etabonate ophthalmic suspension has a better anti-inflammatory effect and less influence on IOP.

Aim To evaluate the bioequivalence of two preparations of gliclazide in healthy volunteers.Methods The concentration of gliclazide was measured by high performance liquid chromatography(HPLC) after a single or multiple dosage of gliclazide sustained released tablet in healthy volunteers.The pharmacokinetic parameters of the two preparations were calculated by 3P97 program.LnAUC0~∞,lnAUC0~72 and lnAUC0~? were used to evaluate the bioequivalence of the two preparations with analysis of variance and two one-side t-test.Results Both the gliclazide extended action tablet were best fitted to one-compartment model.The main parameters of the tested and reference gliclazide after a single dose were as follows:Cmax(2.07?0.61) and(2.26?0.61)mg?L-1;Tmax(5.10?0.55)h and(5.05?0.51)h;T12Ka(1.50?0.26)h and(1.52?0.27)h;T12Ke(8.89?1.56)h and(8.68?1.72)h;MRT(22.63?1.01)h and(22.38?0.93)h;AUC0~72(39.19?8.03)mg?h-1?L-1 and(39.26?8.37)mg?h-1?L-1;AUC0~∞:(45.80?9.51)mg?h-1?L-1 and(45.57?9.76)mg?h-1?L-1;F0~72 and F0~∞(100.19?6.22)% and(100.85?5.88)%,respectively.The main parameters of the tested and reference gliclazide after multiple dose were as follows:Cmax(4.83?0.86)mg?L-1 and(4.69?0.64)mg?L-1;Cmin(0.68?0.14) mg?L-1 and(0.66?0.12)mg?L-1;Tmax:(4.10?0.45) h and(4.10?0.55)h;T12Ka:(2.03?0.53)h and(2.04?0.40)h;T12Ke:(7.24?0.87)h and(7.09?1.14)h;MRT(9.17?0.30)h and(9.19?0.37)h;AUCSS:(41.62?6.48) mg?h-1?L-1 and(42.18?6.03)mg?h-1?L-1;Cav:(1.73?0.27)mg?L-1 and(1.76?0.25)mg?L-1;DF(240.85%?34.07)and(230.23%?24.80%) respectively.The relative bioavailability was(98.60?4.60)%.The AUC0~T,AUC0~∞ or AUCSS,Cmax and Tmax were bioequivalent between the two preparations.Conclusion The tested and reference gliclazide sustained released tablet are bioequivalent.

AIM: To explore the association of single nucleotide polymorphisms of promoter G~ -597 -A, G~ -572 -C, G~ -174 -C and T~ 15 -A, C~ 132 -T in exon 5 of interleukin 6 and lumbar intervebral disc disease. METHODS: The single nucleotide polymorphisms of interleukin 6 gene, including polymorphisms of G~ -597 -A, G~ -572 -C in promoter, G~ -174 -C and T~ 15 -A, C~ 132 -T in exon 5 were analyzed by the polymerase chain reaction and sequencing methods in 81 cases with lumbar intervebral disc disease and 101 healthy controls. The association of single nucleotide polymorphisms of interleukin 6 gene with lumbar intervebral disc disease in two groups was measured. The association of single nucleotide polymorphisms of interleukin 6 gene with lumbar intervebral disc degeneration in those younger than 45-year-old were also measured. RESULTS: The G~ -572 -C polymorphism of interleukin 6 gene was observed, but no single nucleotide polymorphism of G~ -597 -A, G~ -174 -C in promoter and T~ 15 -A, C~ 132 -T in exon 5 in two groups was detected. There was no difference between the distribution of the G~ -572 -C polymorphism of interleukin 6 gene in two groups. In those younger than 45-year-old the association of the single nucleotide polymorphism of interlukin-6 gene and lumbar intervebral disc degeneration was not significant. CONCLUSION: There is G~ -572 -C polymorphism in Chinese. No relation between G~ -572 -C polymorphism of interleukin 6 with lumbar intervebral disc disease and lumbar intervebral disc degeneration was observed.

Objective: To evaluate the effect of "healthy eating plate" based dietary management on diabetic inpatients. Methods: The patients with type 2 diabetes mellitus (T2DM) admitted to Daishan First People's Hospital from November 2019 to November 2020 were selected and randomly divided into two groups. The intervention group was given "healthy eating plate" based dietary management, while the control group was given routine dietary management. Demographic data and physical examination results were collected. Fasting blood glucose (FPG), glycosylated hemoglobin (HbA1c), triglyceride (TG) and total cholesterol (TC) were detected at admission, discharge and 3 months after discharge, and compared between the two groups by covariance and generalized estimating equation. Results: here were 52 patients aged (55.83±9.67) years in the intervention group, with 29 (55.77%) males and 23 (44.23%) females. There were 53 patients aged (57.54±11.09) years in the control group, with 32 (60.38%) males and 21 (39.62%) females. There were no significant differences in FPG, HbA1c, TG and TC levels between two groups at discharge (P>0.05). The level of HbA1c in the intervention group was significantly lower than that in the control group at 3 months after discharge (P<0.05); there were no significant differences in FPG, TG and TC levels (P>0.05). Conclusion The "healthy eating plate" based dietary management can better control the blood glucose of diabetic patients, and can help maintain the dietary treatment. It is worthy of promotion in diabetic patients.

Objective : To investigate the knowledge, attitude and practice on e-cigarette use among college students in Guiyang City, so as to provide insights into tobacco control health education among colleges and universities. Methods College students were sampled from full-time colleges and universities in Guiyang City using a multi-stage stratified cluster sampling method from March to May in 2022. Students' knowledge, attitude and practice on e-cigarette use were collected using a questionnaire designed based on the 2019 China National Youth Tobacco Survey Questionnaire, and factors affecting the attempt to use e-cigarette were identified using a multivariable logistic regression model. Methods: College students were sampled from full-time colleges and universities in Guiyang City using a multi-stage stratified cluster sampling method from March to May in 2022. Students' knowledge, attitude and practice on e-cigarette use were collected using a questionnaire designed based on the 2019 China National Youth Tobacco Survey Questionnaire, and factors affecting the attempt to use e-cigarette were identified using a multivariable logistic regression model. Results : Totally 2 800 questionnaires were recovered, including 2 694 valid questionnaires, with an effective recovery rate of 96.21%. The respondents included 687 males (25.50%) and 2 007 females (74.50%). The total score of knowledge on e-cigarette use was (2.95±1.02) points, and there were low proportions of knowing that e-cigarettes contained hazardous substances, including nicotine and tar (59.06%) and knowing that e-cigarette use may cause diseases (53.27%). The total score of attitudes towards e-cigarette use was (5.09±2.36) points, and 93.50% of respondents did not use e-cigarettes given by companions. There were 132 students with attempts to use e-cigarettes (4.90%), and there were 29 current e-cigarette users (1.08%). Multivariable logistic regression analysis identified age of 20 years and lower (OR=0.438, 95%CI: 0.267-0.719), living in rural areas (OR=0.458, 95%CI: 0.264-0.794), thinking that e-cigarettes are addictive (OR=0.449, 95%CI: 0.217-0.928), and thinking that e-cigarette smoking is harmful (OR=0.263, 95%CI: 0.131-0.527) as factors protecting from e-cigarette use, and monthly living expenses of more than 2 000 Yuan (OR=2.995, 95%CI: 1.135-7.902), cigarette smoking (OR=19.826, 95%CI: 11.385-34.527), and using companions' e-cigarettes (OR=9.141, 95%CI: 5.534-15.101) and thinking that people around me support my use of e-cigarettes (OR=2.673, 95%CI: 1.426-5.013) were risk factors for e-cigarette use among college students. Conclusions There is a low awareness rate of e-cigarette use among college students in Guiyang City, and the majority oppose e-cigarette use, with a low proportion of attempt to use e-cigarettes. Age, source of students, monthly living expenses, e-cigarette smoking, knowledge on e-cigarette use and use of companions' e-cigarettes may affect the attempt to use e-cigarettes.