1.Expression of ATP-binding cassette efflux pump gene Rv1217c-Rv1218c and its association with drug resistance of Mycobacterium tuberculosis
Hao PEI ; Shiliang ZHANG ; Jun LIU ; Yaxin DAI ; Biao HUANG ; Xu WANG ; Mintao HU ; Shougang KUAI ; Ke WANG
Chinese Journal of Infectious Diseases 2012;30(8):454-458
Objective To explore the association of ATP-binding cassette (ABC) efflux pump gene Rv1217c-Rv1218c and the drug resistance of Mycobacterium tuberculosis.Methods A total of 34 Mycobacterium tuberculosis clinical isolates including 24 drug-resistance isolates which were resistant to riffampicin,isoniazid,streptomycin or ethambutol and resistant to at least one second-line antituberculosis drug,and 10 drug-sensitive isolates were involved in this study.The RNA of isolated strains was extracted and then reverse transcribed. Gene expression was performed by real-time polymerase chain reaction (PCR) and data was analyzed by t test and Logistic regression analysis.Results The expressions of Rv1217c in rifampicin-resistant group (2.13 ± 1.89,t =3.44,P<0.01),isoniazid-resistant group ( 1.84 ± 1.86,t =3.16,P< 0.05),streptomycin-resistant group ( 1.86 ±1.96,t=2.78,P<0.05) and ethambutol-resistant groups (3.36±2.35,t=3.04,P<0.05) were all higher than sensitive isolates (0.42 ± 0.31).The expressions of Rv1218c in rifampicin-resistant group (2.54±1.84,t=3.82,P<0.01),isoniazid-resistant group (2.34± 1.84,t=3.72,P<0.01),streptomycin-resistant group (2.15±1.86,t=3.01,P<0.01) and ethambutol-resistant groups (3.78± 1.78,t=4.22,P<0.01 ) were all higher than sensitive isolates (0.65 ± 0.42).The expressions of Rv1217c and Rv1218c in multidrug resistant group were 2.74±2.07 and 3.33± 1.77,respectively,which were both higher than polydrug resistant group (0.79 ± 0.47 and 1.03 ± 0.79,respectively; t =2.91,P<0.05 ; t =3.84,P<0.01,respectively).Logistic regression analysis found that high Rv1217c expression was positively correlated with rifampicin resistance and negatively correlated with isoniazid resistance (both P< 0.01 ),while high Rv1218c expression was negatively correlated with rifampicin resistance and positively correlated with isoniazid resistance (both P<0.01 ).High expressions of two genes were both positively correlated with ethambutol resistance and multidrug resistance (both P<0.01 ) and not statistically correlated with streptomycin resistance (P>0.05).Conclusions The expressions of ABC efflux pump gene,Rv1217c-Rv1218c,are correlated with multiple drug resistance.The overexpression may contribute to the multidrug resistance of Mycobacterium tuberculosis.
2.Effect of lncRNA TUG1 on osteogenic/odontogenic differentiation of human dental pulp stem cells
JIANG Yaxin ; ZHANG Hua ; SUN Linghan ; LI Shiting ; FENG Hao
Journal of Prevention and Treatment for Stomatological Diseases 2022;30(12):844-851
Objective:
To explore the effects of long noncoding-RNA (lncRNA) taurine upregulated gene 1 (TUG1) on the proliferation and osteogenic/odontoblast differentiation of human dental pulp stem cells (hDPSCs).
Methods :
hDPSCs were isolated and cultured. The surface antigens CD44, CD45, CD73, CD90, CD133 and STRO-1 were detected by flow cytometry. Alkaline phosphatase (ALP) staining and alizarin red staining were used to identify the ability of cells to differentiate. RNA was collected on Days 0, 7 and 14 of the osteogenic induction of hDPSCs, and qRT-PCR was used to detect the relative expression of TUG1. The hDPSCs were stably transfected with a lentiviral vector containing the TUG1-silenced pSLenti-U6-shRNA(TUG1)-CMV-EGFP-F2A-Puro-WPRE to silence TUG1. The ability of hDPSCs to proliferate was assessed with the CCK-8 method. ALP and alizarin red staining and quantitative detection were used to detect the ALP activity and formation of mineralized nodules of hDPSCs. The expression levels of dentin sialophosphoprotein (DSPP), dentin matrix protein-1 (DMP-1), Runt-associated transcription factor 2 (Runx2), osteocalcin (OCN) and osteopontin (OPN) genes and proteins were measured by qRT-PCR and Western blot.
Results :
The hDPSCs were successfully isolated and cultured, and TUG1 expression was significantly increased during osteogenic differentiation (P<0.05). The hDPSCs proliferation was suppressed after silencing TUG1(P<0.05). After osteogenic induction, ALP and alizarin red staining showed that ALP activity and mineralized nodules were suppressed by silencing TUG1. The expression levels of the odontogenic differentiation gene DSPP and DMP-1 and the osteogenic differentiation gene Runx2, OCN and OPN were also significantly decreased (P<0.05).
Conclusion
Knocking down TUG1 can inhibit the proliferation and osteogenic/odontogenic differentiation of hDPSCs.
3.Diagnostic values of different diagnostic criteria of 18F-DCFPyL PET/CT for primary prostate cancer
Le MA ; Wanchun ZHANG ; Luoping ZHAI ; Yaxin HAO
Chinese Journal of Nuclear Medicine and Molecular Imaging 2022;42(12):734-738
Objective:To compare the diagnostic values of different diagnostic criteria of prostate specific membrane antigen (PSMA) PET/CT for primary prostate cancer (PCa).Methods:From May 2019 to May 2021, 2-(3-(1-carboxy-5-((6- 18F-fluoro-pyridine-3-carbonyl)-amino)-pentyl)-ureido)-pentanedioic acid ( 18F-DCFPyL) PET/CT images of 78 patients (age: (68.5±1.4) years) with clinically suspected PCa in Shanxi Bethune Hospital were retrospectively collected and blind diagnosed by the three criteria of SUV max, PSMA reporting and data system (PSMA-RADS) score and molecular imaging PSMA (miPSMA) score. The diagnostic efficacy for PCa, the correlation between the diagnostic results and disease risk, and the consistency of the diagnostic results of the three criteria were compared. Delong test, Spearman rank correlation analysis, and intra-class correlation coefficient (ICC) were used to analyze data. Results:The sensitivities of SUV max, PSMA-RADS score and miPSMA score for PCa were all 93.75%(60/64) and the specificities were 12/14, 10/14 and 12/14 respectively; AUCs of the three criteria were 0.951, 0.862 and 0.951, with no significant difference between SUV max and miPSMA score ( z=0.00, P=1.000), while there were significant differences between PSMA-RADS score and SUV max or miPSMA score ( z values: 2.71, 2.93, P values: 0.007, 0.030). There were positive correlations between the diagnostic results of the three criteria and the disease risk (International Society of Urological Pathology (ISUP) grading: rs values: 0.66, 0.62, 0.63, all P<0.001; D′Amico grouping: rs values: 0.67, 0.64, 0.67, all P<0.001). The diagnostic results of the three criteria were highly consistent (ICC=0.941, 95% CI: 0.903-0.967). Conclusion:The SUV max and miPSMA score have higher diagnostic efficiency and correlation of disease risk, which are more suitable for clinical application.
4.Performance evaluation of nucleic acid testing system for blood screening in high-altitude areas
Liang ZANG ; Lei ZHOU ; Ji DUO ; Hao LIN ; Rui BAI ; Mei YANG ; Chao DAN ; Yaxin FAN
Chinese Journal of Blood Transfusion 2023;36(10):935-939
【Objective】 To validate the performance of a nucleic acid testing(NAT) system for blood screening in the high-altitude Nagqu region of Tibet, in order to assess the capability of NAT in high-altitude areas and further enhance blood safety. 【Methods】 Various methods were employed to evaluate the analytical sensitivity, reproducibility, ability to prevent cross-contamination, and comparison between different NAT systems. 【Results】 The NAT system in the Nagqu region of Tibet achieved a 100% detection rate for high-concentration HBV DNA and HIV-1 RNA samples, and over 90% for medium-concentration samples. PROBIT analysis revealed the lower limits of detection (LOD) for HBV DNA and HIV-1 RNA to be 8.29 IU/mL (95% CI, 5.88~20.55 IU/mL) and 40.52 IU/mL (95% CI, 30.26~85.92 IU/mL), respectively. For HCV RNA genotype 2a, the LOD was 97.14 IU/mL (95% CI, 71.00~182.67 IU/mL), all of which were lower than the declared minimum detectable concentrations in the instructions. Reproducibility analysis demonstrated a 100% level of consistency within the system. Cross-contamination performance verification showed a strong ability to resist cross-contamination. Comparative analysis of repeated testing of low-concentration HBV DNA samples and multi-system testing in plain areas revealed consistency rates of 77.78%(14/18) and 77.27%(17/22), respectively, indicating certain differences between the NAT system in Nagqu region and other systems. 【Conclusion】 The NAT system exhibited excellent performance in blood screening at high altitudes. The results of performance validation in high-altitude blood screening NAT systems were largely consistent with those in plain areas, providing a reliable basis for enhancing blood safety in high-altitude regions.
5.Safety evaluation of sintilimab in combination with chemotherapy for the treatment of cholangiocarcinoma
Hao ZHONG ; Hang LIN ; Yaxin LU ; Haiyan MAI
China Pharmacy 2025;36(4):482-485
OBJECTIVE To assess the safety profile of sintilimab in combination with chemotherapy for the treatment of cholangiocarcinoma. METHODS The data of patients with cholangiocarcinoma from January 1st, 2021 to December 31st, 2022 were collected and divided into control group (29 cases) and observation group (18 cases) based on different medication regimens. Patients in the control group were treated with Gemcitabine hydrochloride for injection+Cisplatin for injection or Oxaliplatin for injection, the observation group was treated with Sintilimab injection based on the control group. Patients in each group underwent blood routine, liver and kidney function, biochemical and other examinations before and after each treatment cycle to observe the occurrence of adverse drug reactions. The correlation of adverse drug reactions with drugs was evaluated with Naranjo’s scale. RESULTS The correlation between blood toxicity and drug use was deemed “probable” in both groups; however, the observation group exhibited a significantly higher score, indicating a stronger correlation. In the control group, hepatotoxic reactions were classified as “suspicious” whereas in the observation group, they were categorized as “probable”. The correlation of gastrointestinal symptoms between the two groups was considered “possible”. Systemic symptoms, skin toxicity, musculoskeletal toxicity, endocrine toxicity and renal toxicity were all classified as having a “suspicious” correlation with drug use. The total incidence of blood toxicity in the observation group was significantly higher than control group (P=0.014). There was no statistically significant difference in the total incidences of hepatotoxic, gastrointestinal symptoms, systemic symptoms, skin toxicity, musculoskeletal toxicity, endocrine toxicity, renal toxicity, or the incidence of grade 3 or higher blood toxicity, hepatotoxic between the two groups (P>0.05). For the patients experiencing adverse drug reactions, the symptoms were alleviated following drug discontinuation or symptomatic supportive treatment. No fatalities occurred during the treatment period. CONCLUSIONS Sintilimab combined with chemotherapy may significantly increase the risk of blood toxicity in patients with cholangiocarcinoma, especially thrombocytopenia, but the adverse reactions are within a controllable range, and the overall safety is good.