In the present study, A fluorescent imaging-based high-throughput screening method was developed for identifying anti-migratory compounds with 96-well Transwell plates. The correlation, precision and stability of this method were examined and the incubation time of dye Hoechst 33342 in addition to migration time was optimized. In addition, The inhibitory activity of anti-cancer drug paclitaxel on tumor cell migration was assayed and an IC50 value of 0.717 micromol x L(-1) was obtained. Using this method, 24 components from Rhizoma Alismatis were screened and one component with anti-migration activity was found. These results show that the new proposed method with good precision, stability and linear range has the potential to assay the inhibitory activity of anticancer compounds.

Objective To establish a whole-body dynamic inhalation exposure system for toxicological studies on highly toxic chemicals, and to evaluate the safety and applicability of the system.Methods The safety and standardization of the laboratory were ensured after positive and negative pressure protection and airtight protection were finished.By modifying and optimizing the key technological units of the exposure chamber, the relationships between aerosol concentrations in the chamber and the push rate, exposure time and different monitoring points were investigated.Results and Conclusion Multi-protection was achieved, including the independent exposure chamber, negative pressure experiment and positive pressure protection under working conditions.The laboratory meets the demands of safety and specifications.The exposure aerosol concentrations in the chamber are uniform, stable and controllable while the air is dynamically flowing.The whole-body dynamic inhalation exposure system can meet the need for toxicological studies on highly toxic chemicals.

Objective To evaluate the bacteriostasis of sulfamethoxazole(SMZ)combined with trimethoprim(TMP)against My‐cobacterium tuberculosis(MTB) in vitro ,so as to provide basis for clinical application .Methods The minimal inhibitory concentra‐tion(MIC) value of TMP/SMZ against MTB ,including standard sensitive strain(H37Rv) ,clinical sensitive strains(20 strains) and clinical multiple‐drug‐resistance strains(MDR ,30 strains) ,was detected by using MPP observation method .Results The MIC val‐ue of standard strain H37Rv was TNP(0 .5 μg/mL)+SMZ(9 .5 μg/mL) .The growth of 40 strains(accounted for 80% ) of clinical isolates ,including 17 sensitive strains and 23 MDR strains ,could be inhibited by TMP(1 μg/mL)+ SMZ(19 μg/mL) compound . Conclusion TMP combined with SMZ may has good antibacterial activity for strains of MTB in vitro .

Objective To study the mRNA expression of apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G) in the peripheral blood mononuclear cells (PBMC) in patients with chronic hepatitis C (CHC) and its regulation by exogenous interferon-α (IFN-α).Methods Twenty-eight CHC patients were recruited as case group and 14 healthy subjects were recruited as control group.APOBEC3G mRNA level (the ratio of APOBEC3G mRNA to housekeep geue 18s rRNA) in PBMC was determined by TaqMan real-time polymerase chain reaction (RTPCR).APOBEC3G mRNA levels were also dynamically measured in CHC patients treated with pegylated interferon (IFN)-α 2a at week 0,2,4,12,24,36 and 48 of treatment,and the plasma levels of IFN-α were simultaneously detected by enzyme-linked immunosorbent assay (ELISA).The data were analyzed by t test and analysis of variance using SPSS 11.0 software.Results The level of APOBEC3G mRNA in PBMC of CHC patients before treatment was 1.60× 10-4 ± 1.35 × 10-4,which was significantly lower than healthy controls 6.20 × 10-4 ±1.30 × 10-4 (t=3.147,P=0.003).The expressions of APOBEC3G mRNA were upregulated at week 12,24,36 and 48 of IFN treatment,which were 5.69×10-3±1.61×10-2,1.01×10-2±2.15×10-2,2.01×10-2±3.75×10-2 and 2.45× 10-2 ±4.08× 10-2,respectively,and all higher than that of pretreatment (F=3.46,5.67,10.27 and 25.65,respectively; P=0.042,0.030,0.010 and 0,respectively).IFN-α level in plasma were increased with treatment and reached the plateau at week 2 of the treatment until the end of observation.Conclusion Hepatitis C virus infection may be one of the reasons of APOBEC3G downregulation.

Objective To compare the plasma hepatitis C virus(HCV)RNA levels detected by the fully automated viral load detection system(COBAS TaqMan)and the national real-time quantitative polymerase chain reaction(PCR)kit,and to investigate the clinical application value of these two methods in clinical practice.Methods A total of 168 serial plasma samples collected from 26 patients with chronic hepatitis C(CHC)before and at week 2,4,12,24,36 and 48 of antiviral treatment were detected by both COBAS Taqman 48 analyzing system and the national real-time quantitative PCR kit.The results of two methods were compared by chi square test and t test.Resnlts Both COBAS and national kit showed great positive detecting results when HCV RNA≥1×104IU/mL(at week O),and the virus load value detected by national kit was significantly higher than that detected by COBAS(t=2.05,P<0.05).However,when HCV RNA<1×104(at week 2-48),the positive rate of HCV detected by COBAS was significantly higher than that detected by national kit (t=3.66,P<0.01).At week 4 of treatment,the rapid virological response(RVR)rate was 46.2 % (12/26)detected by COBAS,while that was 88.5%(23/26)detected by national kit,and the difference was significant(x2=10.575,P<0.01).At week 12 of treatment,the complete early virological response(cEVR)was 95.7%(22/23)detected by COBAS,while that was 100%(17/17)detected by national kit,and the difference was not significant(x2=0.726,P>0.05).Conclusions The national TaqMan real-time quantitative PCR kits could be used to screen the suspected cases of HCV infecrion and to diagnose CHC cases with high HCV virus load.COBAS detection is more sensitive in cases with low HCV virus load and in on-treatment monitor during anti-HCV therapy.

Objective To study the relationship between APOBEC3G mRNA level in peripheral blood mononuclear cells (PBMC) and serum hepatitis C viral RNA level in patients with chronic hepatitis C infection. Methods TaqMan real-time fluorescence relative quantitative polymerase chain reaction (RT-PCR) was used to quantify APOBEC3G mRNA levels in PBMC from 49 patients with chronic hepatitis C (CHC) and 31 healthy subjects. The relationship between APOBEC3G mRNA level and hepatitis C virus (HCV) viral load was analyzed. SPSS11. 0 statistics software was used for t test and regression analysis. Results APOBEC3G mRNA level in CHC patients [(1.5×10-5±1.9×10-5 ) copy/mL] was significantly lower than that [( 5. 2 × 10-5 ± 5. 5 × 10-5 ) copy/mL] in the healthy control subjects (t=-3.005, P＜0.01). While APOBEC3G mRNA level was not related with HCV viral loads (r=-0.082, P＞0.05). Conclusion HCV has an inhibitive effect on APOBEC3G expression, whereas APOBEC3G doesn't affect HCV replication directly in vivo.

Objective To detect the change of hepatitis C virus (HCV)RNA in the peripheral blood mononuclear cells (PBMC)and serum of patients with chronic hepatitis C (CHC)during treatment with peg-interferon α-2a (Peg IFNα-2a)plus ribavirin (RBV),and to analyze the clinical significance of HCV RNA detection in PBMC.Methods The peripheral blood samples of 20 CHC patients who visited Department of Infectious Diseases in Guangzhou No.8 People′s Hospital from June 2013 to December 2014,were collected during treatment with Peg IFNα-2a+RBV at different time points (week 0,2,4, 12,24,36 and 48).Serum and PBMC were separated.Accurate fluorescence quantification assay (Cobas TaqMan real time polymerase chain reaction[PCR])was used to detect HCV RNA level in serum,while real-time PCR and nest-PCR were applied to detect HCV RNA in PBMC.Categorical data were analyzed byχ2 test.Results Accurate fluorescence quantification of serum HCV RNA showed that HCV RNA level decline rapidly after treatment (F = 148.06,P < 0.01 ),and 18 patients achieved HCV RNA undetectable at week 12 of treatment.The positive rate of nest-PCR was higher than real-time PCR (all P <0.01).Comparison of HCV RNA levels in serum and PBMC from 20 cases found that,the clearance rate of HCV RNA in PBMC was postponed.Two patients whose HCV RNA in PBMC kept detectable relapsed at week 24 after end of treatment.Conclusions HCV RNA can be detected in PBMC of CHC patients and the positive rate of nest-PCR is higher than real-time PCR.Antiviral therapy is effective on HCV both inside and outside PBMC,but the clearance rate of HCV RNA in PBMC is postponed compared with that in serum.Slow clearance of HCV in PBMC may be a risk factor for relapse after end of treatment.

Objective To explore the quality of life ( QoL ) and muscle strength in patients with pseudohypoparathyroidism ( PHP ) under regular treatment. Methods Twenty-three patients with PHP regularly followed at Peking Union Medical College Hospital from June 2017 to June 2018 were included. Age- and gender-matched 23 patients with nonsurgical hypoparathyroidism ( nHPT) and 23 healthy controls were also included. Short Form 36 Health Survey questionnaire version 2 ( SF36v2) were used to evaluate the QoL. Grip strength and repeated chair stand ( RCS) were used to assess muscle strength for upper and lower limbs respectively. Results Except for physical functioning, patients of PHP group had reduced scores in all other subdomains of SF36v2 compared to healthy controls ( P<0.05) . Comparing to nHPT patients, PHP patients had a higher score in social functioning, while no difference was found in other subdomains of SF36v2. Grip strength and RCS tests were similar in PHP patients and healthy controls. Conclusions Comparing to healthy controls, patients with PHP still had impaired QoL despite regular management, no significant difference of upper and lower limb muscle strength was found between PHP group and healthy controls.

Objective Idiopathic pulmonary fibrosis, the fibrosis score (i.e., the combined extent of reticulation and honeycombing) is associated with worse survival. The aim of this study was to identify high-resolution computed tomography (HRCT) patterns and patient characteristics that could predict poor prognosis in rheumatoid arthritis-related ILD (RA-ILD). Methods We retrospectively analyzed 130 patients with newly diagnosed RA-ILD from 2011 to 2017 at Shanxi People's Hospital. The Pearson correlation analysis was used for the correlation between the fibrosis score and the worse survival of RA-ILD, and Using Cox regression analysis was used to identify the associations with mortality. A value of P less than 0.05 was considered statistically significant. Results During a median follow-up of 65 months, 32/130 (24.6％) patients died. Univariate analysis identified 6 significant poor prognostic factors: lower baseline ％ predicted forced vital capacity [HR=0.97, 95％CI(0.94, 0.99);P=0.008], total interstitial disease score [HR=1.06, 95％CI(1.03, 1.08);P<0.01], reticulation score [HR=1.07, 95％CI (1.04, 1.09); P<0.01], traction bronchiectasis score [HR=2.04, 95％CI (1.21, 3.40);P=0.008], fibrosis score [HR=1.07, 95％CI (1.01, 1.13);P<0.01], and definite UIP pattern [HR=4.18, 95％CI (1.40, 12.51); P=0.010]. Fibrosis score remained to be an independent significant poor prognostic factor of survival on bivariate analysis [HR=8.136, 95％CI (2.87, 28.35); P=0.001]. Patients with a fibrosis score>20％ had high mortality. Conclusion This study has shown that fibrosis score is strongly associated with worse survival in RA-ILD, and patients with fibrosis score>20％ have a 8.136-fold increased risk of mortality.

OBJECTIVE: To establish a rapid method for EBV detection with loop-mediated isothermal amplification (LAMP), and to make it as a clue for early diagnosis of nasopharyngeal carcinoma cancer. METHOD: EBV DNA was fast extracted from samples after boiling, while the whole detection will be finished within an hour with specific amplification of EBV gene by LAMP. RESULT: High specificity was shown from EBV detection of 33 clinical samples. Comparing with PCR, LAMP is more simple and convenient to perform under isothermal conditions, and require no special apparatus, thus, it is more economical and practical. CONCLUSION LAMP analysis of EBV may be an efficient and easy way for clinical diagnosis of nasopharyngeal carcinoma.
DNA Primers ; DNA, Viral ; analysis ; Herpesvirus 4, Human ; genetics ; Humans ; Molecular Sequence Data ; Nasopharyngeal Neoplasms ; diagnosis ; Nucleic Acid Amplification Techniques ; Sensitivity and Specificity