Objective To study curative efficacy of euthyrox in treatment of hypothyroidism during pregnancy and its effect on levels of TSH , FT3 and FT4.Methods 100 patients of hypothyroidism during pregnancy who received therapy from June 2014 to June 2015 in our hospital were selected as research objects, and divited into control group and observation group,50 cases in each group.The control group was treated with Fugui Dihuang wan, while the observation group was treated with euthyrox on the basis of control group.Then the levels of TSH,FT3 and FT4, pregnancy outcome, neonatal adverse outcome, treatment outcome of two groups after treatment were compared.Results After treatment, the level of TSH in observation group was lower than the control group, the levels of FT3 and FT4 was higher than the control group ( P<0.05 ); Incidence of pregnancy complications in observation group was less than the control group [12.00%(6/50)vs 40.00%(20/40)](P<0.05);Total neonatal adverse outcomes [8.00%(4/50) vs 26.00%(13/50)](P<0.05); The total effective rate of observation group was statistically higher than that in the control group [96.00%(48/50) vs 80.00%(40/50)](P<0.05).Conclusion Euthyrox is well for hypothyroidism during pregnancy, which can improve TSH, FT3 and FT4 levels, relieve clinical symptoms.

Objective To investigate the effects of recombinant trichosanthin(rTCS) on methylation status and expression level of p27 gene in HeLa cells.Methods HeLa cells was treated by different concentration(20 ?g/mL,40 ?g/mL,and 80 ?g/mL) of rTCS for 48 h and then methylation-specific polymerase chain reaction(MSP) was used to detect the promoter methylation status of the p27 gene,real-time PCR was used to detect levels of p27 and DNMT1 mRNA,and Western blotting assay was used to detect expression level of p27 protein before and after treatment with rTCS.Results Low expression level and promoter methylation status of the p27 gene were detected in HeLa cells.Treatment with 40 ?g/mL rTCS totally demethylated p27 promoter.Treatment with 20 ?g/mL,40 ?g/mL or 80 ?g/mL rTCS resulted in a 2.22-,4.00-or 6.03-folds increase in p27 mRNA level,respectively,and also a great increase in p27 protein level.A high DNMT1 expression level was observed in HeLa cells and treatment with 40 ?g/mL rTCS resulted in a 78% decrease at the DNMT1 mRNA expression.Conclusion rTCS could reverse promoter hypermethylation and re-activate the expression of p27 gene by inhibiting DNMT1 expression in HeLa cells,which indicates its potential use in cancer therapy.

Hepatitis C virus (HCV), one of the major pathogens of viral hepatitis, causes significant hazards in humans. Interferon treatment in combination with ribavirin is used as the first line clinical treatment for HCV infection. However, good response to this treatment has only been observed in few patients and repeated recurrence has also been reported frequently. Therefore, new antiviral agents and therapies are in urgent demand. Here, we report a newly constructed Escherichia coli RNase P based M1GS ribozyme that can specifically and efficiently target the core gene of HCV. The guide sequence (GS) of this M1IGS was designed according to the sequence of the core coding region of HCV genome. The GS was then covalently linked to the 3' terminus of M1 RNA, the catalytic subunit of RNase P from Escherichia coli. The specification of this sequence-specific ribozyme, M1GS, was then examined using an in vitro cleavage assay. The cytotoxicity and its activity in inhibition of HCV gene expression and viral proliferation were further studied in vivo. Our results show that the reconstructed M1GS ribozyme displayed obvious catalytic activity in cleaving target mRNAs fragment in vitro. Notable reduction in the expression of HCV core protein and a 1 000-fold reduction in viral growth were also observed in cultured HCV infected Huh7.5.1 cells expressing the functional M1GS ribozyme. This study demonstrated a direct evidence for the antiviral activity of the customized M1GS-HCV/C141 ribozyme, and thus provided a promising new strategy for clinical treatment of HCV infection.
Antiviral Agents ; pharmacology ; Escherichia coli ; genetics ; Genetic Engineering ; Hepacivirus ; genetics ; physiology ; RNA, Catalytic ; genetics ; pharmacology ; RNA, Guide ; genetics ; Ribonuclease P ; genetics ; Viral Core Proteins ; genetics

BACKGROUND:Our previous studies have shown that strontium-doped calcium polyphosphate containing low-dose strontium appears to have a significant effect on angiogenesis-related behaviors of monocultured umbilical vein endothelial cells and osteoblasts.
OBJECTIVE:To investigate the effect of strontium-doped calcium polyphosphate on angiogenesis-related behaviors of umbilical vein endothelial cells and osteoblasts co-cultured, including celladhesion, spreading, proliferation, as wel as the protein secretion of vascular endothelial growth factor and basic fibroblast growth factor from co-culture system in vitro.
METHODS:Human umbilical vein endothelial cells and osteoblastic cells (MG63) were utilized in this study. cells from passage 3 were used for preparation of the cel-scaffold constructs. After placed in 24-wel plate at a ratio of 2:1, human umbilical vein endothelial cells and MG63 cells were seeded onto strontium-doped calcium
polyphosphate, calcium polyphosphate and hydroxyapatite scaffolds and co-cultured for 7 days. The vascular endothelial growth factor and basic fibroblast growth factor protein levels were determined through a double ligand enzyme-linked immunosorbent assay. The colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay was performed to quantify the effect of scaffolds on cellproliferation.
RESULTS AND CONCLUSION:Compared with those on calcium polyphosphate and hydroxyapatite scaffolds, cells on strontium-doped calcium polyphosphate scaffolds attached and spread better with a significantly improved cellproliferation. More importantly, the vascular endothelial growth factor and basic fibroblast growth factor expressions were significantly higher in the strontium-doped calcium polyphosphate group than the other two groups (P<0.05), indicating strontium-doped calcium polyphosphate can up-regulate levels of vascular endothelial growth factor and basic fibroblast growth factor proteins.

OBJECTIVE To assess the ultrasonographic features and clinical significance of the complications of the thyroglossal cysts. METHODS We retrospectively reviewed the ultrasonographic findings in 41 cases with complications of the thyroglossal cyst, which was confirmed surgically and pathologically. 43 cases with simple thyroglossal cyst consisted of the control group. The location, size, internal echo, internal septa, wall thickness, posterior acoustic feature, boundary, vascularity, and fistulas of the lesion were analyzed and compared with the control group. RESULTS Most of the inflammatory thyroglossal cyst showed thickening walls, indistinct boundaries, posterior echo enhancement, and peripheral vascularity on ultrasound images. There were significant differences of the thickness of the cyst wall, indistinct boundary, and peripheral vascularity between the two groups. CONCLUSION The characteristic findings of thyroglossal cyst with inflammation are thickened wall, indistinct boundary, and peripheral vascularities. Ultrasonography can be useful tool in determination of the appropriate time of the operation to reduce the recurrence rate.

Objective To investigate the wake-promoting effect of vagus nerve stimulation (VNS) on coma rats after traumatic brain in-jury (TBI), and the related mechanism. Methods A total of 168 healthy Sprague-Dawley rats were randomly divided into blank group, TBI group, antagonist group and VNS group, 42 rats in each group. The latter three groups were established TBI model with impact, and the rats in coma at least 30 minutes were included. VNS group accepted VNS, the antagonist group were injected intralateroventricularly Orexin A receptor 1 (OXR1) antagonist SB334867, and TBI group accepted sham VNS. Their behaviors were observed to determine the level of con-sciousness six, twelve and 24 hours after intervention, while the expression ofγ-aminobutyric acid b1 receptor (GABAb1R) in prefrontal cortex was detected with immunohistochemistry and Western blotting. Results There were 42 rats in the blank group, 11 rats in TBI group, 13 rats in the antagonist group, and 28 rats in VNS group awakened finally. The expression of GABAb1R in prefrontal cortex ranged as TBI group, antagonist group, blank group and VNS group from more to less twelve and 24 hours after intervention under Western blotting (F>60.412, P<0.001), and it ranged as TBI group, antagonist group, VNS group and blank group under immunohistochemistry (H=15.121, P=0.002), with no significant difference among time points (H=3.028, P=0.220). Conclusion VNS can promote waking from coma in rats after TBI, which may relate with the decrease of GABAb1R in prefrontal cortex that induced by Orexin A.

Objective:To investigate the correlation between plexinC1 (PLXNC1) rs2272335 polymorphism and the family clus-tering genetic susceptibility to primary liver cancer (PLC) in Guangxi and the expression of PLXNC1. Methods:Genotype and alleles of rs2272335 were determined in 20 liver cancer family groups (79 cases) and 10 healthy normal control groups (40 cases) in Fusui County through Time of Flight Mass Spectrometer. Immunohistochemistry detected the PLEXNC1 protein expression. Results:For the alleles of PLXNC1 (rs2272335) site, the risk of hepatocellular carcinoma (HCC) for individuals with [C] allele was 4.16-fold (95%CI=0.37-47.3, P=0.032) compared with that for individuals with [T] allele among the members of the healthy normal control group. The fre-quencies of the [C] and [T] alleles were similar in the HCC patients and the core individuals of liver cancer families (P>0.05). For the genotype of the PLXNC1 (rs2272335) site, the differences in frequencies of TT, TC, and CC genotypes were not statistically significant among the PLC patients and the core individuals of the liver cancer families and normal controls. The PLXNC1 protein expression in HCC (3.12±1.12) was higher than in hepatocellular paracancerous tissues (1.54±0.67) and in benign hepatocellular lesions (1.23±0.87) (P<0.05). Conclusion:The [C] allele of PLXNC1 (rs2272335) site might be the risk gene for the occurrence of PLC family clustering in Guangxi. PLXNC1 protein overexpression was closely correlated with PLC oncogenesis.

Objective To construct a prokaryotic expression vector pET-22b+with Middle East respiratory syndrome ( MERS) coronavirus nuclocapsid protein( NP) gene and to express and purify N protein.Methods N gene amplified by PCR was inserted into the prokaryotic expression vector pET-22b+.Recombinant plasmid was confirmed using DNA elec-trophoresis and sequencing.NP was expressed in E.coli BL21(DE3) by IPTG induced and purified by cation exchange chromatography using the AKTA purification system.Results The NP gene sequence was proved to be correct by sequen-cing and the protein was expressed in both soluble and insoluble forms in E.coli BL21 ( DE3 ) after IPTG induction.The purity and concentration of recombinant protein was improved obviously by cation exchange chromatography and enrich-ment.Conclusion Recombiant NP of high purity and concentration is purified and will facilitate NP functional research.

Objective To evaluate the clinical application value of Xpert detection system of Clostridium difficile (C.difficile).Methods A total of 43 stool specimens from the patients with diarrhea were collected,and C.difficile in stool specimens were detected by the Xpert detection system,the toxigenic culture method,and the toxin detection method which detected the toxin of C.difficile by VⅥDAS automatic analyzer after anaerobic culture,respectively.The analytic performance of Xpert detection system was evaluted based on the toxigenic culture method as the gold standard.Meanwhile,the consistency of the results from different detection methods was compared.The ribotype 027 strain (ATCC BAA-1870) simulating the stool specimen was further used to verify the Xpert detection system.Results Based on the gold standard of the toxigenic culture method,the sensitivity,specificity,positive predictive value and negative predictive value of the Xpert detection system were 90.9％,93.8％,83.3％ and 96.8％,respectively.The Kappa values for the consistency between the Xpert detection system and the toxigenic culture method or the toxin detection method were 0.822 (P ＜ 0.05) and 0.419 (P ＜ 0.05),respectively.Moreover,the ribotype 027 strain simulating the stool specimen was verified by the Xpert detection system successfully.Conclusion The Xpert detection system may rapidly and accurately detect the C.difficile in stool specimens,especially the ribotype 027 strain with high toxicity.

Broth microdilution method was hired to measure the minimum inhibitory concentrations (MICs) of luteolin against Trueperella pyogenes,Escherichia coli,Salmonella and Streptococcus in order to evaluate the antimicrobial activity of luteolin in vitro.Meanwhile,the bacteria growth curves in medium containing sub-inhibitory concentration of luteolin were measured in this test.The results indicated that Tureperella pyogenes was the most sensitive to luteolin with a MIC of 0.078 g/L than that of these strains;and Salmonella was also sensitive to luteolin (MIC:1.25 g/L).However,The inhibitory effect of luteolin on Escherichia coli and Streptococcus is relatively weak,and shared the same MIC with 2.5 g/L.Luteolin showed inhibitory effects on the growth curves of all the strains in this test at sub-inhibitory concentration,and the inhibitory effects on the growth curves increased with the concentration of luteolin(P＜0.05).