Objective An HPLC-MS/MS method was established for the simultaneous determination of 7 components in Qingkailing Oral Liquid.Methods The assay was performed on a Waters ACQUITY UPLC BEH C18 column(2.1 mm×10 mm,1.7 μm)and the sample was eluted with a gradient mobile phase containing 10 mmol·L-1 of ammonium acetate and 0.1%of formic acid in water(A)-methanol(B).The mass spectrometry was carried out by electrospray ionization(ESI)with positive/negative ions in multiple reaction monitoring(MRM)mode for quantitative analysis.Results The linear ranges of adenine,chlorogenic acid,caffeic acid,geniposide,baicalin,hyodeoxycholic acid and cholic acid were 0.100 4-3.213,0.784 5-8.982,0.998-3.194,0.622 5-19.92,25.05-300.6,2.513-30.15 and 7.775-93.30 μg·mL-1(r≥0.999 0).The average recoveries(n=6)were 100.9%,98.74%,101.2%,100.2%,100.8%,99.97%and 98.94%with RSD of 1.58%,0.59%,1.78%,1.25%,0.65%,1.69%and 1.07%.The contents of the above mentioned 7 components in 15 tested samples were in the ranges of 0.12-0.18,0.19-0.24,0.06-0.09,0.34-0.37,4.54-4.85,0.49-0.67 and 1.82-2.19 mg·mL-1.The contents of 7 components in tested sample from different manufacturers were closed.Conclusion The method has shown good sensitivity,accuracy,and repeatability.The study can provide reference and data support for the quality control and subsequent research of Qingkailing Oral Liquid.

Humans ; HIV-1/genetics* ; Anti-Retroviral Agents/therapeutic use* ; Mutation/genetics* ; Treatment Failure ; HIV Infections/genetics* ; Drug Resistance, Viral/genetics* ; Anti-HIV Agents/therapeutic use* ; Genotype

Objective:To explore the coexisting gene mutations of FLT3-ITD mutation and its association with partial clinical parameters in acute myeloid leukemia (AML).Methods:The clinical data of 236 newly diagnosed AML outpatients and hospitalized patients of Changzhou No.2 People's Hospital and the Second People's Hospital of Wuxi between December 2012 and August 2019 were retrospectively analyzed. Genome DNA-polymerase chain reaction (PCR) combined with Sanger sequencing was used to detect FLT3-ITD mutations, and 51 tumor target gene mutations in patients with FLT3-ITD mutations were detected by using high-throughput DNA sequencing combined with Sanger sequencing.Results:Among 236 AML patients, FLT3-ITD mutations were found in 71 cases (30.1%). About 97.2% (69/71) patients with FLT3-ITD mutations were accompanied by additional mutations, of which 19 patients harbored double coexisting genes mutations, 24 patients harbored 3 coexisting genes mutations and 26 patients harbored ≥ 4 coexisting genes mutations. The most common coexisting genes mutations were NPM1 (55 cases, 77.5%), followed by DNMT3A (36 cases, 50.7%), TET2 (9 cases, 12.7%), CEBPA (5 cases, 7.0%), IDH1 (4 cases, 5.6%) and NRAS (4 cases, 5.6%). In FLT3-ITD mutation group, the hemoglobin level of patients with DNMT3A mutation type was lower than that of those with DNMT3A wild type ( t = -2.37, P = 0.020); the hemoglobin level of patients with NPM1 mutation type was higher than that of those with NPM1 wild type ( t = 2.04, P = 0.045). The platelet in patients with 3 mutations and ≥ 4 mutations was higher than that in those with double mutations ( χ2 = 7.72, P = 0.021). After chemotherapy in 71 patients, the curative effect of 66 cases was evaluable, and the white blood count of 18 patients who did not reach complete remission was higher than that of 48 patients who reached complete remission ( Z = -2.74, P = 0.006). Conclusions:Most FLT3-ITD mutated patients with AML commonly show coexisting gene mutations, and the mutation types of coexisting genes are correlated with the clinical features of patients.

Metformin is currently a strong candidate anti-tumor agent in multiple cancers. However, its anti-tumor effectiveness varies among different cancers or subpopulations, potentially due to tumor heterogeneity. It thus remains unclear which hepatocellular carcinoma (HCC) patient subpopulation(s) can benefit from metformin treatment. Here, through a genome-wide CRISPR-Cas9-based knockout screen, we find that DOCK1 levels determine the anti-tumor effects of metformin and that DOCK1 is a synthetic lethal target of metformin in HCC. Mechanistically, metformin promotes DOCK1 phosphorylation, which activates RAC1 to facilitate cell survival, leading to metformin resistance. The DOCK1-selective inhibitor, TBOPP, potentiates anti-tumor activity by metformin in vitro in liver cancer cell lines and patient-derived HCC organoids, and in vivo in xenografted liver cancer cells and immunocompetent mouse liver cancer models. Notably, metformin improves overall survival of HCC patients with low DOCK1 levels but not among patients with high DOCK1 expression. This study shows that metformin effectiveness depends on DOCK1 levels and that combining metformin with DOCK1 inhibition may provide a promising personalized therapeutic strategy for metformin-resistant HCC patients.
Animals ; Antineoplastic Agents/therapeutic use* ; Carcinoma, Hepatocellular/metabolism* ; Cell Line, Tumor ; Clustered Regularly Interspaced Short Palindromic Repeats ; Genome ; Humans ; Liver Neoplasms/metabolism* ; Metformin/therapeutic use* ; Mice ; Phosphorylation ; Synthetic Lethal Mutations ; Transcription Factors/metabolism* ; rac GTP-Binding Proteins/metabolism*

Objective To study the relationship among bile components and different gallstone types through comparing and analyzing gallbladder bile contents in patients with different types of gallstones.Methods A retrospective study of 542 consecutive patients with gallstones or gallbladder polyps was conducted.The stone composition type and 14 kinds of bile components from these patients were analyzed.The bile parameters consisted of potassium (K +),sodium (Na+),chlorine (Cl-),calcium (Ca2+),bicarbonate (HCO3-),magnesium (Mg2+),aspartate aminotransferase (AST),glutamyltranspeptidase (GGT),alkaline phosphatase (ALP),total bilirubin (TBIL),total bile acid (TBA),cholesterol (CHO),lactate dehydrogenase (LDH) and pH.Finally,the content of these bile components among the different types of stones and gallbladder polyps were compared.According to the composition determined by Fourier transform infrared spectroscopy (FTIR),the gallstone patients were divided into five groups.Results Compared with other groups,the content of K +,GGT,ALP,TBIL,TBA and CHO in the calcium carbonate stone group were lower (P ＜ 0.05),while the levels of C1-,HCO3-and value of PH were higher (P ＜0.05).Furthermore,CHO content in the cholesterol stone group was higher than other groups (P ＜ 0.05)except for the gallbladder polyp group (P ＞ 0.05).In addition,there was no difference in bile contents among the pigment gallstone group,mixed stone group and polyp group (P ＞ 0.05).Conclusions In gallstone patients,the bile components of patients with calcium carbonate stones is significantly different.The high cholesterol content in bile is the main feature of cholesterol stone patients,and there is no significant difference in bile composition between patients with pigment stones and mixed stones.

OBJECTIVE： To provide reference for improving emergency capacity of the hospital pharmacy department in response to the novel coronavirus pneumonia （COVID-19） epidemic. METHODS ：According to the related regulations and requirements of Law of the People ’s Republic of China on the Prevention and Control of Infectious Diseases ，combined with the situation of COVID- 19 epidemic prevention and control ，and management experience of relevant hospitals ，on the basis of in-depth analysis of drug supply and quality assurance ，drug dispensing management ，provision of clinical pharmaceutical services and other related material support of hospital pharmacy department，integrated emergency management model was constructed for COVID- 19 epidemic prevention and control ，and the precautions and response measures of each link were sorted out. RESULTS ：Integruted emergency management mode for COVID-19 epidemic prevention and control in hospital pharmacy department included but was not limited to human resource management，drug and disinfection products supply management （mainly including key treatment drugs and disinfection product list formulation，control，inventory increase ，etc.）；drug dispensing management （mainly including prescription ，pharmacy window ， planning quantitative reserve ， drug return ， etc.）；clinical pharmaceutical care management （mainly including providing pharmaceutical information support ，online pharmaceutical service ，monitoring drug safety ，etc.）；personnel protection and disinfection （mainly including personnel protection ，environment and window ，equipment and container ，paper prescription disinfection，etc.）；special management of donated drugs ；prevention and control knowledge training ；pharmaceutical education and scientific research management ，etc. CONCLUSIONS ：The integrated emergency management model for epidemic prevention and control is helpful for hospital pharmacy to manage public health emergencies. During the outbreak of COVID- 19，hospital pharmacy department should start integrated emergency management mode for epidemic prevention and control ，strengthen the risk control of each link ，and play a good role in the key functional departments in the special period.

Objective:Biobank construction plays an irreplaceable role in the research of accurate prevention and treatment of diseases. Shared biobank network based on a large crowd queue is the way of the future. This subject is one of the key contents of national precision medicine "The Breast Cancer Cohort Study in Chinese Women: (BCCS-CW)" , aiming to solve the bottleneck of insufficient standardization and sharing.Methods:The establishment of "entity library-information library-extension library" , the widely Shared network of biobank of breast cancer specific disease cohort, and the establishment of strict standard setting and quality control standard to construct the standardized biobank.Results:This biobank provides a shared biobank resource for breast cancer risk assessment, prediction and early warning, early screening, classification, individualized treatment, efficacy and safety prediction and monitoring and other accurate prevention and treatment programs and clinical decision-making system research.Conclusion:The data of this biological sample bank is refined and complete, and the sample size of cases is sufficient, which can meet the research needs of medical big data, genomics, metabonomics, epigenetics and other fields.

Objective To detect ASXL1 gene variants among patients with myelodysplastic syndrome (MDS) and explore their correlation with variants of other genes and clinical features of patients.Methods For 149 patients with MDS,genomic DNA was amplified by PCR and subject to direct sequencing to identify variants of ASXL1,U2AF1,SF3B1,DNMT3A,TET2,IDH1/2,NPM1,FLT3-ITD and C-KIT genes.Results ASXL1 variants were found among 37 patients (24.8％).Other commonly mutated genes included U2AF1 (22.8％),TET2 (11.4％),DNMT3A (9.4％),NPM1 (8.1％) and SF3B1 (6.0％).The frequency of concurrent U2AF1 and TET2 variants among patients with ASXL1 variants was slightly higher than that of wild-type patients.No significant difference was found in median age,MDS subtype,karyotype,peripheral leukocytes,hemoglobin,platelet levels,and bone marrow blast counts between the ASXL1-variant and the wild-type groups (P＞0.05).Twenty-nine patients harboring ASXL1 variants were followed up,37.9％ progressed to acute myeloid leukemia (AML).The rate of transformation in ASXL1-variant group was significantly higher than the wild-type group (37.9 ％ vs.14.1％,P＜0.01).Conclusion ASXL1showed a high frequency of variant among MDS patients,which was frequently accompanied with U2AF1 and TET2 variants.Compared with the wild type group,patients with ASXL1 variants were more likely to progress to AML.

Objective: To investigate the clinical value of the serum new molecular markers, soluble triggering receptor expressed on myeloid cells-1(sTREM-1)and soluble hemoglobin scavenger receptor(sCD163), in the diagnosis of sepsis in elderly patients with burns. Methods: A total of 58 inpatients with burns from Jun 2017 to June 2018 were enrolled in the study.Patients were divided into three groups: the sepsis group(n=12), the localized infection group(n=21)and the non-infection group(n=29). The levels of sTREM-1 and sCD163 were determined by enzyme-linked immunosorbent assays(ELISAs). The clinical diagnostic value of sTREM-1 and sCD163 was assessed by receiver operating characteristic(ROC)curve analysis. Results: There was a statistically significant difference in the levels of sTREM-1 and sCD163 at day 1 between the three groups(F=20.994 and 38.363, P<0.01). Serum levels of sTREM-1 and sCD163 were higher in the sepsis group than in the localized infection group and the non-infection group.Serum levels of sTREM-1 and sCD163 were higher in the localized infection group than in the non-infection group.Serum levels of sTREM-1 and sCD163 were lower at day 7 than those at day 1 in all groups(F=21.242 and 41.035, P<0.01). Serum sTREM-1 levels were positively correlated with serum sCD163 levels(r=0.609, P=0.000). The AUC of sTREM-1 and sCD163 for the diagnosis of sepsis was 0.880(95%CI: 0.816~0.926). Conclusions Serum levels of sTREM-1 and sCD163 are elevated with increasing degrees of infection.Monitoring serum sTREM-1 and sCD163 levels is helpful for the diagnosis of sepsis in elderly patients with burns.

Objective: To carry out mutation analysis for patients with myelodysplastic syndromes (MDS) and a normal karyotype. Methods: Targeted capture and next-generation sequencing (NGS) was carried out using a customized 49-gene panel. FLT3 internal tandem duplication (FLT3-ITD), CALR, NPM1 and CEBPA mutations were detected by PCR and Sanger sequencing. Results: Sixty two patients (80.5%) were found to harbor at least one mutation. Each patient has carried 2.21 mutations in average. Coexistence of ≥ 3 mutations was common (43.7%). The most commonly mutated genes were RUNX1 (23.4%, 18/77), ASXL1 (18.2%, 14/77), NPM1 (15.6%, 12/77), U2AF1 (15.6%, 12/77), DNMT3A (11.7%, 9/77). Patients with SF3B1 mutations were significantly older than those with ASXL1 mutations (P=0.023). Mutations of the DNMT3A gene were significantly associated with the blood platelet level compared with BCOR mutations (P=0.02). No significant difference was found in the number and rate of mutations between those under or above 60-year-old. Among 67 patients with clinical follow-up, 20 (29.8%) has transformed to acute myeloid leukemia, and the time of transformation has ranged from 1 to 44 months, with a average of 5.3 months. RUNX1, U2AF1 and FLT3 mutations are associated with leukemic transformation. Conclusion Coexistence of ≥ 3 mutations are frequent among patients with normal-karyotype MDS. Certain mutations are associated with age and leukemic transformation.