OBJECTIVE: To establish a HPLC method for the determination of content of entecavir and its related substances. METHODS: The HPLC condition was consisted of a Luna C18 column with a column temperature at 25 ℃, using gradient eluate method, the mobile phase was acetonitrile,with a flow rate of 1.0 mL?min-1 and detection at 254 nm. RESULTS: The calibrated linear curve of entecavir was within 0.033 45～0.167 2 mg?mL-1(r=0.999 98). The average recovery was 100.09%,RSD=0.20%, The content of related substance was 0.54%～1.48%. CONCLUSION: This accurate and reliable HPLC method is applicable for the quality control of entecavir and its related substances.

Objective To investigate the effects of changes of miR-126 and spouty related EVH,domain containing proteinl(SPRED1) after transient ischemic attack(TIA)on prognostic value for pathogenesis of secondary cerebral infarction.Methods Retrospective analysis of the clinical data of 106 patients with TIA was performed.The expression levels of miR-126,SPRED1 and vascular endothelial growth factor(VEGF)in peripheral blood were detected at 3 h,6 h and 12 h after TIA onset respectively.The specificity and sensitivity of miR 126 and SPRED1 in the diagnosis of TIA were analyzed.The miR-126 and SPRED1 levels versus ABCD2 score were compared for evaluating their predictive value in the diagnosis of secondary cerebral infarction within 30 days after TIA onset.Results The miR-126 level was declined after TIA onset at 3 h(9.41±1.04),especially at 12 h(6.59 ±2.78),versus in healthy control (9.35±1.76)(t =-7.764,P=0.000).The SPRED1 level after TIA onset was increased at 3 h(58.05 ± 17.53)pg/L,12 h(82.64 ± 18.60)pg/L versus in healthy control(52.38 ± 13.24)pg/L(t=12.374,P =0.000).A closely negative correlation was found between levels of miR 126 and SPRED1 at 12 h point but not at 3 h and 6 h(r=-0.278,P=0.004).Both miR-126 and SPRED1 levels at 12 h after TIA were implied to sensitivity and specificity evaluation.Additionally,VEGF was significantly increased at 3 h (345.61 ± 76.76) pg/L,6 h (461.65 ±103.87)pg/L and 12 h (519.22 ± 103.55)pg/L after TIA onset as compared with healthy control (107.77± 26.04) pg/L(t =26.569,29.756,34.699,all P =0.000).The decrease of miR-126 and increase of SPRED1 at 12h after TIA indicated high incidences of cerebral infarction but their significance was less than ABCD2 score.Combination of miR 126,SPRED1 and ABCD2 score significantly improved the prediction for cerebral infarction(Z=2.105,P =0.035).Conclusions After the onset of TIA,levels of miR-126 and SPRED1 expression in combination of ABCD2 score can improve predictive value for cerebral infarction development.

OBJECTIVE:To establish an RP-HPLC method for content determination of s-omeprazole sodium and its related substances.METHODS:The separation of s-omeprazole sodium and the related substances was carried out on a Phenomenex Luna C18 column,the mobile phase consisted of methanol-0.033 mol?L-1 ammonium dihydrogen phosphate-triethylamine (58∶41.8∶0.2,adjusted to pH 7.0 by phosphate acid).The detection wavelength was 302 nm,the flow rate was 1.0 mL?min-1,the column temperature was 25 ℃,and the sample size was 20 ?L.RESULTS:The linear range of omeprazole sodium was 10～500 mg?L-1 (r=0.999 7).The average recovery rate was 100.27% (RSD=0.74%).The average content of the related substances in samples was 0.42%.CONCLUSION:This method is simple,accurate,specific and applicable for content determination of s-omeprazole sodium and its related substances.

OBJECTIVE:To establishment the method for the determination of content of vitamin A palmitate(VAP) and its related substances by HPLC. METHODS:The HPLC conditions were consisted of Phenomenex Luna C18 column with a mobile phase of a mixture of acetonitrile-isopropanol (90∶10) ,the detection wavelength of 328 nm,the column temperature of 30 ℃ and the flow rate of 1.0 mL?min-1. RESULTS:VAP was completely separated from impurities,the linearity range was 90～400 mg?L-1(r=0.999 2). The average recovery rate was 99.60% (RSD=1.32%). The average content of the related substances were lower than 2.53% . CONCLUSION: This accurate and reliable HPLC method is applicable for the quality control of VAP.

OBJECTIVETo investigate the drug resistance of HIV patients to the HIV-1 CRF01_AE and CRF07_BC strains in Sichuan province during 2010 to 2013.

METHODS1.5 ml of plasma were collected from AIDS patients who had been receiving anti-retroviral treatment for over 6 months but still had a HIV-1 virus load of over 1 000 copies/ml from January 1, 2010 to December 31, 2013 in Sichuan province. Genetic analysis of the HIV-1 pol gene was performed using self-established method, and patients with a positive drug-resistant HIV-1 pol gene mutation were included. HIV-1 poly gene was successfully sequenced for a total of 1 213 patients. Drug resistance of different HIV-1 strains was compared with χ2 test or Fisher exact test.

RESULTS558 cases (46.0%) of the 1 213 successfully sequenced patients were infected by HIV-1-strains with drug-resistant mutations, including 327 cases (58.6%) infected by CRF01_AE strain, 126 (22.6%) by CRF07_BC strain, 46 (8.2%) by CRF08_BC strain, 33 (5.9%) by B strain, 4 (0.7%) by C strain, 1 (0.2%) by CRF02_AG strain, and 21 (3.8%) by unidentified strains. Drug-resistant mutation analysis revealed that L33, F116, L74, Q151, and T69 resistance mutations occurred only in the CRF01_AE strain, while A71, K43, and Q58 resistance mutations occurred only in the CRF07_BC strain; in nuclear nucleoside reverse transcriptase inhibitors (NRTIs) and non nucleoside reverse transcriptase inhibitors (NNRTIs), CRF01_AE subtype strains showed highly resistant rate were higher than CRF07_BC, CRF08_BC and B subtype strains, with the differences were statistically significant (P<0.05).

CONCLUSIONThe drug-resistant HIV-1 strains in Sichuan mainly included the CRF01_AE and CRF07_BC strains, which had different resistance mutations.

Base Sequence ; Drug Resistance, Viral ; Genes, pol ; HIV Infections ; HIV-1 ; Humans ; Mutation ; Reverse Transcriptase Inhibitors ; Viral Load

Objective: To evaluate the efficacy and safety of intravenous thrombolysis with recombinant tissue-plasminogen activator (rt-PA) in elderly patients with early-stage mild ischemic stroke (IS). Methods: This was a prospective, open-label, controlled study.Ninety-four elderly patients with mild IS admitted to our hospital from January 2014 to December 2017 were randomized into a thrombolysis arm (TA, n=46) and a control arm (CA, n=48). The short-term endpoints were the National Institutes of Health stroke scale (NIHSS) scores on 3^rd, 7^th, 14^thday after admission and the secondary endpoints were the modified Rankin Scale (mRS) score and the morbidity of recurrence IS within 90 days.Safety was evaluated by the incidence of intracranial hemorrhage (IH) and early neurological deterioration (END) during hospitalization. Results: The baseline NIHSS scores of patients in the TA and CA groups were similar [(4.1±0.7) vs.(4.1±0.7)]. However, there were significant differences in the NIHSS score on 3^rd [(3.4±1.2) vs.(4.2±1.4)], 7^th[(3.0±1.8) vs.(4.1±1.6)] and 14^thday [(2.5±2.0) vs.(3.4±1.6)], respectively, between the TA group and the CA group.Furthermore, the TA group was associated with a significantly higher proportion of patients with good prognosis (mRS, 0-2), compared with the CA group (71.7% vs.35.4%, P<0.01). Receiver operating characteristic curve analysis showed that patients with baseline NIHSS>3 could benefit from thrombolytic therapy.There were 1 case of symptomatic IH and 1 case of progressive stroke in the TA group, and 1 case of IH and 2 cases of progressive stroke in the control group.There were no significant differences in the rate of either END or IH between the two groups (P>0.05). Two patients in the TA group and three patients in the control group had recurrent IS within 90 days and the recurrence rate of IS was also similar within 90 days (P>0.05). Conclusions Intravenous thrombolytic therapy with rt-PA can improve the prognosis of elderly patients with mild stroke without increased risk of END, IH, or recurrence of IS.

Objective: To construct and screen optimal siRNA interference sequence of CIT gene and to detect its interference efficiency as well as proliferation effect in human hepatoma cell line SK-Hep-1. Methods: Three siRNA target spots were designed and synthesized according to the CIT gene sequence. SK-Hep-1 HCC cells were transfected by liposome transfection. The knockdown efficiency of the target CIT gene was detected by real-time PCR and Western blot. Expressional change of CIT in SK-Hep-1 cells after 48 hours of siRNA interference were observed by immunohistochemistry and confocal microscopy. The proliferation of SK-Hep-1 cells after 48 hours of siRNA interference was detected by EdU cell proliferation assay. A t-test was used to compare the mean of two samples, and one-way ANOVA was used to compare the mean of multiple samples. Results: Western blot results showed that the three interference sequences were targeted at different target spots. The expression level of CIT protein in KD-1,-2, and-3 groups were decreased (P < 0.01) than control, while the protein expression level of KD1 group was the lowest. Real-time PCR results showed that compared with the control group, the expression level of CIT mRNA in KD-1, -2, and -3 groups decreased (P < 0.01), while that in KD1 group was the lowest. Laser confocal microscopy also confirmed that the morphological expression of CIT attenuated significantly after transfection with siRNA. The results of EdU proliferation assay showed that siRNA transfected with CIT significantly attenuated the proliferation of SK-Hep-1 hepatoma cells (P < 0.05). Conclusion The successful construction and screening of siRNA fragments can effectively inhibit the expression and proliferation of CIT gene in hepatoma SK-Hep-1.

Objective: To investigate the impact of transforming growth factor-β1 (TGF-β1) silencing by small interfering RNA (siRNA) on the expression of platelet-derived growth factor-β (PDGF-BB) and its receptor (PDGF-βR) in rats with hepatic fibrosis. Methods: A total of 40 male Sprague-Dawley rats were randomly divided into normal control group, model group, TGF-β1 siRNA treatment group, and negative control group. All rats except those in the normal control group were given subcutaneous injection of 40% carbon tetrachloride to establish a rat model of hepatic fibrosis. The rats in the negative control group and the TGF-β1 siRNA treatment group were given tail vein injection of negative control plasmid or TGF-β1 siRNA plasmid twice a week at a dose of 0.25 mg/kg, and those in the normal control group and the model group were given the injection of sterile isotonic saline twice a week. The rats were sacrificed after 12 weeks and liver tissue samples were collected. Real-time PCR, immunohistochemistry, and Western blot were used to measure the expression of PDGF-BB, PDGF-βR, and phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) in liver tissue. A one-way analysis of variance, the q test, and the Kruskal-Wallis test were used for statistical analysis based on data type. Results: Compared with the model group and the negative control group, the TGF-β1 siRNA treatment group had significantly inhibited mRNA and protein expression of PDGF-BB and PDGF-βR (F = 24.785 and 22.92, P < 0.01), as well as significantly inhibited expression of p-ERK1/2 (P < 0.05). Conclusion Targeted TGF-β1 siRNA can effectively downregulate the expression of PDGF-BB, PDGF-βR, and p-ERK1/2 in liver tissue and thus help to improve hepatic fibrosis.

Humans ; Noninvasive Ventilation ; Cannula ; Airway Extubation ; Retrospective Studies ; Pulmonary Disease, Chronic Obstructive/therapy* ; Hypercapnia/therapy* ; Oxygen Inhalation Therapy ; Respiratory Insufficiency/therapy*