Objective:To establish HPLC fingerprint of Shaoyao Gancao Keli to provide quality control.Methods:Waters Xselect CSH C18 column was used.The mobile phase was acetonitrile-0.05％phosphoric acid solution with gradient elution.The detection wavelength was 232 nm.The column temperature was 35 ℃.The flow rate was 0.8 mL·min-1.The injection volume was 10 μL.10 samples were tested by the method.The similarity of 10 samples were evaluated,and the common peaks were identified.Results:HPLC fingerprint of 17 common peaks was estab-lished.10 common peaks were identified which were gallic acid,albiflorin,paeoniflorin,liquiritin apioside,liquiritin,β-1,2,3,4,6-pentagalloylglucose,isoliquiritin apioside,isoliquiritin,liquiritigenin,glycyrrhizic acid.The similarity of 10 samples was higher than 0.95.Conclusion:The method has good separation,accuracy,re-peatability and stability,and could be used as a standard for quality control of Shaoyao Gancao Keli.

Objective To analyze the characteristics of HIV-1 pretreatment drug resistance(PDR)and molecular transmission networks in Yubei District,Chongqing,providing evidence for targeted interventions.Methods Using a cross-sectional design,plasma samples were collected from HIV/AIDS patients receiving antiretroviral therapy(ART)in Yubei District from January 2022 to December 2023.Pol gene fragments were extracted and amplified for HIV-1 genotyping and drug resistance analysis.Molecular transmission networks were constructed based on genetic distance calculations.Results Among 478 HIV-1 pol sequences,eight geno-types were identified:with CRF07_BC(60.4%,289/478),CRF08_BC(15.5%,74/478),CRF01_AE(11.7%,56/478),and CRF85_BC(5.9%,28/478).The overall PDR rate was 6.3%(30/478),with resistance to nucleoside reverse transcriptase inhibitors(NRTIs)and non-nucleoside reverse transcriptase inhibitors(NNRTIs)at 1.7%(8/478)and 5.2%(25/478),respectively.No protease inhibitor(PI)resistance was de-tected.The molecular network included 177 cases(37.0%network entry rate),forming 53 clusters with 198 connections.Cluster sizes ranged from 2 to 17 nodes,and 75.3%(149/198)of connections were associated with five subdistricts/towns:Shuanglonghu Street,Huixing Street,Luoqi Town,Gulu Town,and Baoshenghu Street.Conclusion HIV-1 genotypes in Yubei District exhibit diversity and complexity,with moderate PDR prevalence.Regional clustering of transmission networks suggests the need for enhanced molecular surveil-lance and targeted interventions based on analytical findings.

Objective:To establish HPLC fingerprint of Shaoyao Gancao Keli to provide quality control.Methods:Waters Xselect CSH C18 column was used.The mobile phase was acetonitrile-0.05％phosphoric acid solution with gradient elution.The detection wavelength was 232 nm.The column temperature was 35 ℃.The flow rate was 0.8 mL·min-1.The injection volume was 10 μL.10 samples were tested by the method.The similarity of 10 samples were evaluated,and the common peaks were identified.Results:HPLC fingerprint of 17 common peaks was estab-lished.10 common peaks were identified which were gallic acid,albiflorin,paeoniflorin,liquiritin apioside,liquiritin,β-1,2,3,4,6-pentagalloylglucose,isoliquiritin apioside,isoliquiritin,liquiritigenin,glycyrrhizic acid.The similarity of 10 samples was higher than 0.95.Conclusion:The method has good separation,accuracy,re-peatability and stability,and could be used as a standard for quality control of Shaoyao Gancao Keli.

Objective : To investigate the effects of melatonin ( MEL) on the proliferation of hepatic stellate cells (HSCs) induced by platelet - derived growth factor (PDGF⁃BB) and explore its correlation with the regulation of autophagy levels . Methods : The HSC⁃T6 cells were divided into the following groups : control group , model group and MEL (low , medium and high) treatment groups . After 24 hours culture , the cells adhered to the wall and were changed into serum⁃free DMEM medium to synchronize the cells to the G0 stage . After 24 hours culture , all groups were given with PDGF ⁃ BB ( 10 ng/ml) excepted the control group . Besides , melatonin of different concentrations ( 1 nmol/L , 1 μmol/L and 0. 1 mmol/L) were added immediately in three treated groups . After incubated for 48 hours , the effect of MEL on the proliferation of hepatic stellate cells activated by PDGF⁃BB was detected by CCK8 method . The protein expression levels of LC3b and α ⁃SMA in hepatic stellate cells were determined by Western blot . The expression levels of LC3b mRNA and α ⁃SMA mRNA in hepatic stellate cells were determined by qRT⁃PCR . The ultrastructure of HSCs was observed by transmission electron microscopy to understand the autophagy level. Results : Compared with control group , PDGF⁃BB could induce the proliferation of HSCs ( P < 0. 01) . Compared with model group , MEL inhibited the proliferation of HSCs activated by PDGF⁃BB ( P < 0. 01) . Compared with the control group , LC3b and α ⁃SMA protein expressions significantly increased in the model group ( all P < 0. 05) , and LC3b mRNA and α ⁃SMA mRNA expressions significantly increased in the model group (P < 0. 05 , P < 0. 01) . Compared with the model group , MEL could inhibit such effects (LC3b : P < 0. 05 , P < 0. 01 ; α ⁃SMA : P < 0. 01) . Transmission electron microscopy ( TEM) showed that compared with the control group , autopolysosome significantly increased in the model group (P < 0. 05) . Compared with model group , autopolysosome significantly decreased in MEL treatment group (P < 0. 01) . Conclusion The up⁃regulation of autophagy level can promote the proliferation of hepatic stellate cells and the inhibition of hepatic stellate cell proliferation by MEL may be related to the down⁃regulation of autophagy level .

Objective:To analyze the immunotherapy and clinical characteristics of coronavirus disease 2019 (COVID-19) patients, and focus on exploring the effects of immunotherapy and mesenchymal stem cells (MSC) transplantation in the critically ill patients' treatment.Methods:Fity-five COVID-19 patients were admitted to the Fifth People's Hospital of Wuxi from January 23rd to March 31st, 2020 as the research object. The demographic characteristics of the cases and the methods of immunotherapy were analyzed, focusing on the immunized indicators, positivity of pathogens and clinical indicators of critically ill COVID-19 patient, and the effects of immunotherapy and stem cell transplantation were evaluated.Results:Aged, male and people with comorbidities were the main risk factors in the development of severe and critical COVID-19. All of confirmed COVID-19 cases (n = 55) had been treated with interferon-α (IFN-α), of which 81.8% (n = 45, mild and ordinary) of the patients were recovered, 14.6% (n = 8) of the patients were converted to severe, 3.6% (n = 2) of the patients were converted to critical, and some severe patients were treated with gamma globulin and albumin as adjuvant treatment. Critically ill patients were not only treated with IFN-α, gamma globulin and albumin, but also treated with convalescent plasma and MSC transplantation. Due to pulmonary hemorrhage and persistently low blood oxygen saturation, terminal lung transplantation therapy was implemented. The total number of lymphocytes, CD4 +, CD8 + T lymphocytes, natural killer (NK) cells and B cells in peripheral blood of the two critical COVID-19 patients were significantly reduced, and the functions of lung, liver, and kidney were severely damaged on admission, manifested as significant increase of the levels of blood C-reactive protein (CRP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and blood urea nitrogen (BUN), etc. and decrease of blood oxygen saturation, and type Ⅰ respiratory failure, and the noninvasive assisted ventilation was needed to improve. After adjuvant immunotherapy such as gamma globulin, the nucleic acid of 2019 novel coronavirus (2019-nCoV) turned into negative. The CRP of one critically ill patient was significantly lower than the value at admission (minimum of 21 mg/L). But the lung inflammation progressed rapidly, and the pathological results of the lung tissue from the lung transplantation showed hemorrhage and irreversible fibrosis. The ability to secrete immunoglobulin A (IgA) was significantly reduced. Liver function had been significantly improved and stabilized after treatment with convalescent plasma during the recovery period. MSC transplantation treatment reduced the BUN level by > 50% compared with the previous period, and the total number of lymphocytes in the patient increased by more than 2 times (rose from 0.23×10 9/L to 0.57×10 9/L), but the total amount of lymphocytes was still lower than the normal reference value (< 1.1×10 9/L). The lung inflammation lesions were obviously absorbed, and the vital signs were stable. Conclusions:In addition to IFN, gamma globulin, antiserum and MSC transplantation therapy can help clear the virus and reduce inflammation. Although MSC transplantation fail to completely change the immunecompromised state of critically ill patients, it controlled the progression of inflammation in the liver and kidneys.

Objective:To explore the safety and nosocomial infection control measures of COVID-19 patients of end-stage respiratory failure after lung transplantation.Methods:Lung transplantation was performed for a COVID-19 patient with end-stage respiratory failure after a negative conversion of 2019-nCoV nucleic acid. Before operation, all medical staff received simulated training on nosocomial infection. The procedures were performed in an operation room with a negative pressure environment. The three-grade preventive strategy was implemented and wearing positive pressure protective mask of electric air supply required. During operation, the patient was managed according to the in-hospital protection process. The environment and medical instruments were disinfected after operation.Results:The operation was completed successfully. The 2019-nCoV nucleic acid test was negative for many times after operation. Participants were placed in medical isolation for 14 days after surgery. During the period, nucleic acid test was negative twice.Conclusions:As an exploratory treatment, lung transplantation is a safe option for end-stage respiratory failure in COVID-19 in operation room with a negative pressure environment and implementations of three-grade preventive strategy.

Objective To cultivate and improve specialty clinical nursing skills and level of orthopedic nurses by applying standardized patients in their examination, and to explore effective mode of professionalized and standardized training of orthopedic nurses so as to improve nursing quality of orthopedic departments. Methods 3 nurses were recruited as SP and were trained. Examination range was defined. 20 nurses working in department of joint surgery for more than 6 months were examined with SP as objects. Application effect of SP was evaluated.Results Authenticity scores of SP were (3.97±0.79) points. Examination results of nurses with application of SP:nursing diagnosis (2.38±0.66), problem-solving ability (2.37±0.71), communication skills (2.56±0.72) and clinical skills (2.34±0.61), remaining to be improved.Conclusions Application of SP in professional training of clinical nurses in orthopedic departments can improve practice operational ability of nurses, which shows a new direction in in-job training for clinical nurses.