OBJECTIVE: To optimize the formula and preparation technology of gel-matrix sustained release tablet of nicotinic acid(GSTNA).METHODS: The formula of GSTNA was optimized by orthogonal experiment with the amount of hydrophilic gel-matrix material HPMC(K15M,E15-LV) and that of adjuvant calcium hydrogen phosphate(CHP) as factors and with the in vitro release rates as index.Meanwhile,the verification test on the intra-and inter-batch release rates of the samples was performed.RESULTS: The optimum formula could be seen as follows: the ratios of HPMC(K15M,E15-LV) and CHP were 4%,40% and 25% respectively.The GSTNA prepared in this formula achieved a sustained drug release of up to 12 h,and both the intra-batch homogenicity and the inter-batch reproducibility were satisfactory.CONCLUSION: The GSTNA is reasonable in formula and simple in preparation technology.

Objective To establish and evaluate the CaV1?1?R528H gene knock?in mouse model of thyrotoxic hy?pokalemic periodic paralysis. Methods Thirty?six 8?week?old male CaV1?1?R528H gene knock?in mice and thirty?six 8?week?old wild?type male C57BL/6J mice were used in this study. Using three?factor two?level 2 × 2 × 2 factorial design ( the three factors including mutation, thyroxine and insulin, and two levels were with or without) , the mice were divided into 8 groups. The thyroxine groups were intraperitoneally injected with levothyroxine in a dose of 350 μg/kg once per day for 12 consecutive days to produce thyrotoxicosis. The insulin groups were intraperitoneally injected with short?acting insulin in a dose of 0?8 U/kg after the last administration of levothyroxine, and the potassium levels of different groups were meas?ured and recorded before (0 min) and after insulin injection (30 min, 60 min). Results (1) Compared with the control group, the following phenomena including irritability, dull coat, increased diet and water intake, and slow body weight gain, were observed in the thyrotoxic mice. Thyroid function tests showed that the levels of T3 and T4 in the thyrotoxic mice were significantly higher than those in the corresponding control mice (P<0?05), and the TSH level was significantly low?er than that of the corresponding control mice (P<0?05 ). (2) After administration of insulin or thyroxine alone, the po?tassium levels in the mutant and wild?type mice were not significantly different. However, after combined administration of thyroxine and insulin, the potassium levels in the mutant group were significantly lower than those in the wild?type mice at 30 min and 60 min ( P<0?05 for both). (3) The main effects and interactions:Mutation factor or thyroxine factor alone did not influence on the potassium level, only insulin showed hypokalemic effect (P<0?05). There were interactions be?tween thyroxine and mutation, and between insulin and mutation (P<0?05), but no interaction between thyroxine and in?sulin. Conclusions (1) A thyrotoxicosis state in mice is successfully developed in this study. (2) An CaV1?1?R528H gene knock?in mouse model of thyrotoxic hypokalemic periodic paralysis is successfully established.

Objective To investigate the effect of recipient's pre-transplant triglyceride (TG) abnormalities on early graft function (EGF) after kidney transplantation.Methods According to the inclusion and exclusion criteria,154 identified living-kidney transplant recipients in the 309 Hospital of Chinese PLA from Jan.2011 to Dec.2014 were enrolled in present study,including 124 males and 30 females,and aged of 31.9 ± 8.4 years.The cohort was divided into two groups:TG normal group (0.40＜TG≤1.70mmol/L,n=107) and TG abnormalities group (TG＞l.70mmol/L or require lipid lowering therapy,n=47).The incidences of poor early graft renal function (PEGF),slow graft function (SGF) and delayed graft function (DGF) were compared between the two groups,and then the serum creatinine (Scr) levels were compared among the patients showing immediate graft function (IGF) at 3rd,7th and 30th day after transplantation.The ROC curve was drawn up taking TG as diagnosis index to explore the optimal cut-offvalue for predicting PEGF,SGF and DGF after transplantation.Results Compared with the TG normal group,the TG abnormalities group showed significantly higher incidence of PEGF and DGF (P＜0.05).Among the IGF patients,the TG abnormalities group showed higher Scr level at the 7th and 30th day after transplantation (P＜0.05).The area under ROC curve (AUC) reflected TG levels for PEGF,SGF and DGF were 0.774,0.704 and 0.818,respectively (P＜0.05).The optimal cut-offvalues were all 1.37mmol/L.Conclusions Recipients with abnormal pre-transplant TG level may have worse EGF after renal transplantation.The risk of developing PEGF,S GF and D GF tends to emerge when pre-transplant TG level is higher than 1.37mmol/L.

Objective To investigate the single-nucleotide polymorphisms of PTPN22 gene rs2476601 ,rs3811021 and rs2488457 in patients with primary immune thrombocytopenia(ITP) .Methods Totally 100 patients with ITP and 100 cases as con-trol from Department of Hematology ,the Affiliated Baiyun Hospital of Guiyang Medical College and the Affiliated Hospital of Guiyang Medical College were collected .PTPN22 gene + 1858 loci (rs2476601) and 3′UTR region rs3811021 loci were detected by PCR-RFLP ,the promoter-1123 loci (rs2488457) were detected by PCR-SSP ,and the results were statistically analyzed .Results PTPN22 gene + 1858 locus in ITP patients and control group were all C allele ,T allele was detected ,and there was no single nucle-otide polymorphisms (R620W) exist .The frequency of PTPN22 gene rs3811021 locus TT ,CT ,CC three genotypes in ITP patients and control group had no significant difference(χ2 = 3 .686 ,P= 0 .158) .The frequency of T allele ,C allele in ITP patients and con-trol group had no significant difference(χ2 = 2 .828 ,P = 0 .093) .The frequency of PTPN22-1123 gene (rs2488457)GG ,GC ,CC three genotypes in ITP patients and control group had no significant difference(χ2 = 1 .802 ,P = 0 .406) .The frequency of C allele and G allele in ITP patients and control group had no significant difference(χ2 = 0 .003 ,P = 0 .954) .According to the gender fac-tors ,in females ,the genotype and allele frequency of SNP loci rs3811021 and rs2488457 in ITP patients and control group had no significant difference(P< 0 .05) ,so as in males(P < 0 .05) .Conclusion PTPN22 gene rs2476601 this SNP site does not exist in Guizhou Han population ,The addition of two SNP loci of PTPN22 gene (rs3811021 ,rs2488457) exists polymorphism ,but the two SNP loci has no sex difference ,the onset and ITP in Guizhou Han population had no significant correlation .

Objective:To explore the trans-membrane signaling mechanism of interleukin-6 (IL-6)-induced osteogenic differentiation and calcification of human umbilical artery smooth muscle cells (HUASMCs).Methods:HUASMCs were primarily cultured in vitro and were stimulated with IL-6, IL-6+solutable IL-6 receptor (sIL-6R), IL-6+sIL-6R+solutable gp130 (sgp130), or vehicle (blank control). Alizarin red and Von Kossa staining were used for detecting cell calcification, Western blot was used to test the protein expression of tissue-nonspecific alkaline phosphatase (TNAP), osteopontin (OPN), bone morphogenetic protein-2 (BMP-2) and Runt related transcription factor 2 (Runx2), and immunofluorescence was used to examine the mIL-6R expression of HUASMCs. The comparison of measurement date between the two groups was conducted by t-test. The comparison of measurement date between multiple groups was conducted by one-way analysis of variance (ANOVA). Results:The intensity severity of calcification stain was IL-6+sIL-6R group >IL-6+sIL-6R+sgp130 group>IL-6 group=blank control. After stimulated for 12 hours, the TNAP expression in blank control, IL-6 group, IL-6+sIL-6R group, IL-6+sIL-6R+sgp130 group were (0.44±0.08), (0.52±0.14), (0.84±0.16) and (0.55±0.10) respectively ( F=290.96, P<0.001). After stimulated for 3 days, the OPN expression in blank control, IL-6 group, IL-6+sIL-6R group, IL-6+sIL-6R+sgp130 group were (0.61±0.84), (0.95±0.16), (1.65±0.24) and (0.99±0.10) respectively ( F=507.72, P<0.001). After stimulated for 12 hours, the BMP-2 expression in blank control, IL-6 group, IL-6+sIL-6R group, IL-6+sIL-6R+sgp130 group were (0.77±0.05), (1.69±0.16), (2.81±0.26) and (0.57±0.12) respectively ( F=959.09, P<0.001). After stimulated for 3 days, the Runx2 expression in blank control, IL-6 group, IL-6+sIL-6R group,IL-6+sIL-6R+sgp130 group were (0.57±0.03) , (0.92±0.10), (1.31±0.13) and (0.66±0.06) respectively ( F=1141.27, P<0.001). Comparing with Jurkat cells (positive control) and CEM cells (negative control), HUASMCs limited expressed mIL-6R. Conclusion:IL-6 may induce HUASMCs osteogenic differentiation and calcification mainly via the sIL-6R-mediated trans-signaling pathway.

Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies inducedin vivo through hybridoma.Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.ResultsAfter 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×10⁴ U and 33.22×10⁴ U) was higher than that of BALB/c mice (5.33×10⁴ U and 19.31×10⁴ U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×10⁴ U/g) was higher than that of BALB/c mice (0.23×10⁴ U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (bothP＜0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation.