1.In Vitro and in vivo Component Identification of Danshenyin Based on UPLC-Q-TOF-MS/MS
Sitong ZHANG ; Xianrun HU ; Wenkang LIU ; Jinchun LEI ; Xuemei CHENG ; Xiaojun WU ; Wansheng CHEN ; Manlin LI ; Changhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):175-183
ObjectiveTo elucidate the chemical composition of Danshenyin and its blood components in rats after oral administration. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) coupled with PeakView 1.2 software was used to systematically characterize and identify the components of Danshenyin aqueous extract and its migratory components in rat blood after oral administration based on the retention time, quasi-molecular ion peaks, secondary fragmentation ions, and literature reports, and a preliminary compounds identification of Salviae Miltiorrhizae Radix et Rhizoma aqueous extract, the co-decoction of Santali Albi Lignum and Amomi Fructus was carried out to attribute the chemical constituents of the aqueous extract of Danshenyin. ResultsA total of 73 compounds, including 21 phenolic acids, 23 diterpenes, 6 flavonoids, 7 organic acids, 3 volatile oils and 13 others, were identified from the aqueous extract of Danshenyin. And 36 prototypes and 15 metabolites were identified in rat plasma, the major metabolic pathways included reduction, hydration, hydroxylation, demethylation, methylation, sulfation and others, these metabolites were mainly derived from tanshinones and salvianolic acids. ConclusionThe main blood components of the aqueous extract of Danshenyin are salvianolic acids and tanshinones, which may be the material basis of the efficacy. This study can provide reference for pharmacological research, quality control, and clinical application of Danshenyin.
2.Remodeling characteristics and construction of a survival prediction model based on enhancers and regulome in intestinal type gastric cancer
Xu CHEN ; Zhaole CHU ; Bijun QIN ; Biying LIU ; Xianfeng LI ; Tao WANG ; Wenkang LIU ; Bin WANG
Journal of Army Medical University 2024;46(7):695-704
Objective To explore the genome-wide distribution of histone H3K27ac in intestinal type gastric cancer,analyze remodeling features of enhancers and regulome and construct a prediction model for prognosis.Methods H3K27ac CUT&Tag sequencing and RNA sequencing were performed in intestinal type gastric cancer tissues from 15 patients and normal gastric mucosa tissues from 18 healthy volunteers.Bioinformatics analysis was performed to identify the differences in genome distribution of H3K27ac modifications.Based on the distribution characteristics of H3K27ac,the enhancer elements were identified and the remodeling characteristics of enhancer and related regulome were explored.The prediction model for prognosis based on enhancer related target genes was constructed by univariate Cox and multivariate Cox regression analyses.Results The histone H3K27ac modification was mainly distributed in the enhancer region and displayed no significant differences in the genomic distribution patterns between normal and cancer tissues.Compared with normal gastric mucosa,the level of enhancer H3K27ac modification was higher in intestinal type gastric cancer.A total of 8847 enhancers with increased activity in intestinal type gastric cancer were identified,accounting for 8.3%of all enhancers,which might promote malignant behaviors such as proliferation and adhesion of gastric cancer cells.A prognosis-predicting model established based on a panel of 6 genes that upregulated by the acquired enhancer in cancers,which was able to predict the overall survival of patients.Conclusion Enhancer remodeling is one of the significant epigenetic features of intestinal type gastric cancer.These enhancers may drive malignant growth and adhesion of cancer cells by upregulating the expression of MYC,E2F3 and other genes.A prognosis model based on enhancer target genes is constructed.
3.Clinical efficacy of neoadjuvant immunochemotherapy versus neoadjuvant chemotherapy for local advanced gastric adenocarcinoma
Lei WANG ; Linghong WAN ; Tao WANG ; Zhongyi QIN ; Fan LI ; Wenkang LIU ; Bin WANG
Journal of Army Medical University 2024;46(20):2313-2321
Objective To evaluate the therapeutic efficacies of neoadjuvant immunochemotherapy (NAIC )and neoadjuvant chemotherapy (NAC )for local advanced gastric adenocarcinoma and analyze the clinicopathological characteristics.Methods A retrospective cohort study was conducted on 243 patients with locally advanced gastric cancer admitted in Army Medical Center of PLA (Daping Hospital)and the First Affiliated Hospital of China Medical University from January 2017 to July 2023. After balancing the confounding factors by inclusion and exclusion criteria and propensity score matching (PSM),the tumor pathological regression (TRG)rate and safety of the 2 neoadjuvant treatment regimens were analyzed,and the clinical pathological characteristics were analyzed to find clinical pathological characteristics related to efficacy.Results After using PSM to balance the baseline characteristics of the 2 groups of patients,53 subjects in each group were included in the analysis.In terms of TRG,the pathological complete response (pCR)rate in the NAIC group (13 patients,25%)was significantly higher than that in the NAC group (2 patients,3.8%,P<0.05 ).Similar results were observed in terms of major pathological response (MPR),with 23 patients (43%)in the NAIC group achieving MPR,while 9 patients (17%)in the NAC group achieved MPR (P<0.05 ).In terms of safety,the incidence of treatment-related adverse events (TRAEs) of any grade in the NAIC group and the NAC group was comparable (96.2% and 96.2%,respectively).In an exploratory subgroup analysis of tumor pathological regression,the patients with clinicopathological features such as age<65 years,male,stage Ⅲ~Ⅳ A of American Joint Committee on Cancer (AJCC)staging,histological type of adenocarcinoma,high-moderate differentiated,intestinal-type gastric cancer,stage T3~4 of clinical T-staging,and stage N2~3 of clinical N-staging were more likely to benefit from NAIC.Conclusion NAIC results in a higher rate of pathological regression and a comparable incidence of adverse events when compared with chemotherapy alone for patients with local advanced gastric adenocarcinoma.
4.Role and mechanism of metformin in algesia of rats with type 2 diabetic neuropathic pain
Yong SUN ; Wu LIU ; Wei ZHANG ; Fei YU ; Wenkang LUAN ; Xiaoxue ZHAO ; Fucheng DAI
Chinese Journal of Neuromedicine 2021;20(12):1194-1203
Objective:To investigate the role and mechanism of metformin in algesia of rats with type 2 diabetic neuropathic pain (DNP).Methods:Eighty sprague-dawley rats were randomly divided into normal control group ( n=15) and high-fat and high-glucose group ( n=65); normal diet and high-fat and high-sugar diet were given, respectively; before and 8 weeks after feeding, the body mass of rats and fasting blood glucose level were recorded, fasting insulin level was detected by ELISA, and insulin sensitivity index (ISI) was calculated. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) 8 weeks after feeding (baseline values) were measured in the high-fat and high-glucose group; after 12 h of fasting, intraperitoneal injection of streptozotocin (STZ, 35 mg/kg) was performed; 3 d after fasting, blood glucose was measured; 14 d after STZ injection, body mass was recorded and MWT and TWL were measured again: when MWT and TWL were ≤85% baseline values, it was defined that DNP model was successfully established ( n=45); and the left were into the diabetic painless group ( n=15). The rats with successful DNP were randomly divided into DNP group, DNP+vehicle group and DNP+metformin group ( n=15); 14 d after STZ injection, rats in the DNP+metformin group were given intraperitoneal injection of metformin (200 mg/kg) once daily for 14 consecutive d; DNP group did not accept any treatment, and rats in DNP+vehicle group were intraperitoneally injected with same amount of normal saline. MWT and TWL of all rats were measured 14 d after STZ injection, and 3, 7, 14 and 21 d after metformin injection. The expression levels of interleukin (IL)-6, IL-1β and tumor necrosis factor (TNF)-α were detected by ELISA 7, 14 and 21 d after metformin injection. The fluorescence intensity of ionized calcium binding adaptor molecule-1 (Iba-1) in the spinal cord was detected by immunofluorescence staining, and the expression levels of Toll-like receptor 4 (TLR4), nuclear transcription factor (NF)-κB, phosphorylated (p)-NF-κB, adenylate activated protein kinase (AMPK), p-AMPK, and peroxisome proliferator activated receptor-γ coactivator (PGC)-1α in the spinal cord were detected by Western blotting 21 d after metformin injection. Results:(1) After 8 weeks of feeding, the body mass of rats in the high-fat and high-glucose group was significantly higher than that in the normal control group ( P<0.05); and the body mass of rats in the high-fat and high-glucose group was statistically lower than that in the normal control group 14 d after STZ injection ( P<0.05). Three d after STZ injection, the blood glucose level in high-fat and high-glucose group was significantly higher than that in normal control group ( P<0.05). After 8 weeks of feeding, the insulin level of high-fat and high-glucose group was statistically higher than that of normal control group, and the ISI in the high-fat and high-glucose group was significantly decreased as compared with that in the normal control group ( P<0.05). (2) As compared with those in the normal control group and diabetic painless group, MWT and TWL of DNP group and DNP+vehicle group were significantly decreased at each time point ( P<0.05). Three, 7, 14 and 21 d after metformin injection, MWT and TWL in DNP+metformin group were significantly increased as compared with those in DNP group and DNP+vehicle group ( P<0.05). (3) Seven, 14, and 21 d after metformin injection, the levels of IL-6, IL-1β and TNF-α in the spinal cord of rats in the DNP group and DNP+vehicle group were significantly increased as compared with those in the normal control group and diabetic painless group ( P<0.05); as compared with those in the DNP group and DNP+vehicle group, the levels of IL-6, IL-1β and TNF-α in the spinal cord of DNP+metformin group were significantly decreased ( P<0.05). (4) As compared with normal control group and diabetic painless group, the fluorescence intensity of Iba-1 and number of Iba-1 positive cells in the spinal cord tissues of DNP group and DNP+vehicle group were significantly increased ( P<0.05); while the fluorescence intensity of Iba-1 and number of Iba-1 positive cells in spinal cord tissues of DNP+metformin group were significantly decreased as compared with those in the DNP group and DNP+ vehicle group ( P<0.05). (5) As compared with those in the normal control group and diabetic painless group, the TLR4 and p-NF-κB protein expressions and p-NF-κB/NF-κB values in the spinal cord tissues of DNP group and DNP+vehicle group were significantly increased ( P<0.05); while those in the spinal cord tissues of DNP+metformin group were significantly decreased as compared with those in the DNP group and DNP+vehicle group ( P<0.05). As compared with those in the normal control group and diabetic painless group, the PGC-1α protein expression and p-AMPK/AMPK values in the spinal cord tissues of DNP group and DNP+vehicle group were significantly decreased ( P<0.05); while those in the spinal cord tissues of DNP+metformin group were significantly increased as compared with those in the DNP group and DNP+vehicle group ( P<0.05). Conclusion:Metformin, by activating AMPK/PGC-1α signaling pathway, may inhibit the TLR4/NF-κB expression, reduce the activation of microglia and the expressions of pro-inflammatory factors, and thus alleviate DNP.
5.Study on the influencing factors of HIV testing awareness among Myanmar people in Dehong, Yunnan province
Wenkang ZHANG ; Fei DIAO ; Guowu LIU ; Mengjie HAN ; Yuecheng YANG ; Chengbo WANG ; Ximei XIE ; Xiaobin CAO
Chinese Journal of Experimental and Clinical Virology 2021;35(2):130-134
Objective:To analyze the influencing factors of HIV-testing awareness of Myanmar people in Dehong, and provide references for early discovery of HIV infection among Myanmar people.Methods:A cross-sectional study was conducted among 400 Myanmar people in Dehong through convenient sampling method in a form of face-to-face questionnaire interview. HIV-testing awareness and influencing factors were described and analyzed with χ2 and logistic regression method . Results:A total of 393 participants were included in the study, in whom 241 (61.3%) were males, 256(65.1%) were Myanmar people, 233(59.3%) received 1-7 years of education in Myanmar, 348 do not speak Chinese (88.5%), 226(57.5%) were married, 378(96.2%) were non-solitary, 257(65.4%) were workers, the main purpose of etering into China in 273(69.5%) was working in China, 273(69.5%) had stayed in China for more than 2 years, 573(95.4%) had a clear plan on whether to stay in China for a long time in the future. In these Myanmar people, the rate of AIDS-related knowledge awareness was 75.6%; 165(42.0%) had sexual behavior in the last 6 months. 221(56.2%) knew that they had been tested for HIV. Ninety (22.9%) were more likely to actively seek HIV-testing after they had unprotected behaviors such as homosexual sex, injecting drug use, and sex without using condom. Multivariate logistic regression analysis showed that the influencing factors of HIV-testing awareness included being women (compared with men, OR=2.155, 95% CI: 1.088-4.270), educational level of 8-10 years or more than 10 years (compared with education level 1-4 years, OR=5.207, 95% CI: 2.094-12.950; OR=19.780, 95% CI: 5.800-67.457), having sexual behavior in the last 6 months (compared with those who had not, OR=2.534, 95% CI: 1.343-4.779), having received AIDS-related education in the last 6 months (compared with those who had not, OR=2.462, 95% CI: 1.303-4.654), AIDS-related knowledge awareness (compared with those who had no awareness, OR=8.126, 95% CI: 2.936-22.491). Conclusions:Myanmar people have low awareness of HIV-testing in Dehong. Those who were women and high literacy levels, having received AIDS-related knowledge education in the past 6 months, and having AIDS-related knowledge have an active HIV-testing awareness, after they have high-risk behaviors. AIDS-related knowledge should be publicized to promote early discovery of high-risk behaviors.
6.Analysis on influencing factors on HIV testing behaviors in some foreigners in Guangzhou
Si CHENG ; Zhigang HAN ; Fanghua LIU ; Honghua CAO ; Huifang XU ; Yan LI ; Wenkang ZHANG ; Xiaobin CAO
Chinese Journal of Epidemiology 2020;41(7):1092-1096
Objective:To analyze the influencing factors of HIV testing behavior among foreigners in Guangzhou, and provide a reference for promoting HIV testing and intervention services in foreigners.Methods:A cross-sectional survey was conducted from November 2019 to January 2020 among 230 eligible foreigners selected in Guangzhou through convenient sampling method in the form of one-to-one questionnaire interview, HIV testing behaviors and related influencing factors were described and analyzed with χ2 test and multivariate logistic regression model. Results:A total of 230 foreigners were included in the study, in whom 156 were males (67.8%), 217 were from Africa (94.3%), 138 (60.0%) were with education level of university and above, 169 (73.5%) were in business, 47 (20.4%) were students, 132 (57.4%) had cumulative residence >12 months in Guangzhou and 113 (49.1%) had health examination in China in the past 6 months. In these foreigners, the rate of AIDS-related knowledge awareness was 76.0%, the proportion of condom use in the last sex was 39.6%(89/225) and 47.8%(110/230) received HIV tests in the past 3 months. Multivariate logistic regression analysis showed, the influencing factors of HIV testing behavior in the past 3 months included cumulative residence ≤12 months (compared with cumulative residence >12 months, OR=3.12, 95 %CI: 1.49-6.56), health examination in China in the past 6 months (compared to those who had not, OR=2.53, 95 %CI: 1.17-5.48), awareness of HIV testing and consulting service in Guangzhou (compared with those who had no awareness, OR=3.65, 95 %CI: 1.71-7.78). Conclusions:Some foreigners in Guangzhou had relatively low rates of HIV testing and condom use. Those who pay attention to their own health status and know well about HIV testing and consulting services are more likely to receive HIV testing, foreigners in Guangzhou should be encouraged to receive HIV testing. Their self-testing awareness should be improved.
7.Application of T-SPOT.TB in the Early Diagnosis of Tuberculosis
Ruijie SHI ; Wenkang LIU ; Yun FAN ; Bo LI ; Rong LIU ; Junyan WANG ; Ling LI ; Futang YAN
Journal of Modern Laboratory Medicine 2017;32(2):60-63
Objective This study aims to explore the application value of tuberculosis T lymphocytes enzyme-linked immune SPOT test (T-SPOT.TB) on early diagnosis of tuberculosis.Methods The TB infection in 189 inpatients suspected tuberculosis in pneumology department of Shaanxi Provincial People's Hospital was detected with T-SPOT.TB,fluorescence RQPCR,tuberculosis (TB-Ab)protein chip and PPD methods.Results The sensitivity of four methods was 91.54% (119/130),73.85%(96/130),63.08%(82/130) and 57.69% (75/130) respectively and the specificity of those was 89.83% (53/59),86.44%(51/59),67.79%(40/59) and 66.10%(39/59),respectively.The sensitivity of T-SPOT.TB method was statistically higher than those of other three tests,respectively (P<0.05).The specificity of T-SPOT.TB was significantly higher than those of TB-AB and PPD (P<0.05),but there was no statistical difference between RQ-PCR and T-SPOT.TB (P>0.05).The positive predictive values of T-SPOT.TB,fluorescent quantitative PCR,TB-Ab and PPD assays were 95.2% (119/1250),92.3% (96/104),81.2% (82/101) and 78.9% (75/95) respectively while the negative predictive values of those were 82.8% (53/64),60% (51/85),45.5% (40/88) and 41.5% (39/94),respectively.The false-positive rates (misdiagnosis rate) of four assays were 10.2% (6/59),13.6% (8/59),32.2% (19/59) and 33.9% (20/59) respectively and the false-negative rates (rates of missed diagnosis) of those were 8.5% (11/130),26.2% (34/130),36.9% (48/130)and 42.3 % (55/130),respectively.The negative likelihood ratios of T SPOT.TB,fluorescent quantitative PCR,TB-Ab and PPD assays were 0.11,0.16,0.48 and 0.51 respectively,meanwhile the positive likelihood ratios of T-SPOT.TB,fluorescent quantitative PCR,TB-Ab andPPD assays were 9.0,5.4,2.0 and 1.7,respectively.What' s more,the diagnostic accordance rates of the four assays were 91.0% (172 189),77.8% (147 189).64.6% (122/189) and 60.3% (114/189),respectively.Conclusion T-SPOT.TB test is a more sensitive and specific method and of great significance to the early diagnosis of TB,which has more clinical value in different stages of tuberculosis diagnosis.
8.Drug Resistance of Pathogens in Blood Culture Specimens in Shaanxi Antimicrobial Resistant Investigation Net from 2013 to 2015
Qiaodi GUI ; Wenkang LIU ; Jinrong CANG ; Yanyan GONG ; Jiankang REN
Journal of Modern Laboratory Medicine 2017;32(2):89-91
Objective To explore the Gram-positive bacterial drug resistance in blood culture in 2013 to 2015 from the members of Antimicrobial Resistant Investigation Net of Shaanxi province,and guide the clinicians touse antimicrobial drugs rationally.Methods All the objective bacterial isolates were collected and identified susceptibility date by software WHONET 5.6.Results 8 824 Gran positive bacterial isolates and their antibacterial susceptibilitydata were collected.The top five populations of Gram-positive bacterial isolates were Staphylococcus epidermidis,Staphylococcus hominis,Staphylococcus haemolyticus,Staphylococcus aureus and Enterococcus feacium.The isolating rates of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase negative Staphylococcus(MRCNS) were 33.9 ~ 54.9% and 72.1 ~88.6 % respectively.No vancomycin,Linezolid and teicoplanin resistant Staphylococcus isolates were found.There were 0.9 ~2% E.faecium vancomycin-resistant isolates.Conclusion The composition of blood culture from 2013 to 2015 was not changed,The rate of MRSA and MRSE showed downward trend.But it was severe that the situation of bacterial drug resistance in blood culture in Shaanxi province.Should fully use bacterial drug resistance surveillance results for supervision and administration,and take effective measures for controlling the spread of resistant isolates.
9.TORCH Serological Screening and Infection Characteristic in Neonate in Tianjin Area
Zhenzhong LIU ; Fengqi WU ; Jing YAN ; Yan HUANG ; Wenkang LIU
Journal of Modern Laboratory Medicine 2017;32(4):133-136
Objective The purpose of this study was to explore the infection characteristic of Toxoplasma gondii (TOX),Rubella virus (RV),Cytomegalovirus (CMV) and Herpes simplex virus Ⅱ type (HSV-Ⅱ) (TORCH) infection in neonate in Tianjin area.Methods TOX-IgM/IgG,RV-IgM/IgG,CMV-IgM/IgG and HSV-Ⅱ-IgM/IgG were detected in serum of 2 273 neonate during 2015~2016 with enzymelinked immunosorbent assay (ELISA).Results The positive rates of TOX IgM,RV-IgM,CMV-IgM and HSV-Ⅱ-IgM were 0.00%(0/2 273),0.00%(0/2 273),0.88%(20/2 273) and 0.00%(0/2 273),respectively and those of TOX-IgG,RV-IgG,CMV-IgG and HSV-Ⅱ-IgG were 3.65% (83/2 273),86.45% (1 965/2 273),95.82%(2 178/2 273) and 8.27%(188/2 273),respectively.There was 0.66% percent (15/2 273) of examinees who were infected by none of TORCH pathogens.There existed significant statistical difference for positive rate between TOX-IgG,RV-IgG,CMV-IgG and HSV-Ⅱ-IgG (x2 =6.747,P =0.000) with consequence of the highest positive rate being CMV-IgG.The positive rates of TOX-IgG and CMV-IgM in neonate of 2016 were significantly less than those in 2015 (x2 =5.789~7.505,P=0.006~0.016) but that of HSV-Ⅱ-IgG of 2016 was statistically higher than that in 2015 (x2 =6.073,P =0.014).The positive rate of CMV-IgM in male neonate in 2015 was significantly higher than that in 2016 (x2 =5.054,P =0.025).As a whole the positive rates of TOX-IgG,RV-IgG,CMV-IgG and HSV-Ⅱ IgG had no differences between different years,so did those between gender groups (x2 =2.23~6.963,P=0.073~0.526).The positive rates of TOX-IgG,RV IgG,CMV-IgG and HSV-Ⅱ-IgG in female neonate in 2015 were statistically different from those in 2016 (x2 =8.247,P =0.041).The female neonate in 2015 had higher infection proportion of TOX-IgG compared with that in 2016 (x2 =6.992,P=0.008).TORCH infection detected in 2 273 cases of neonate had one pathogen infection and multi-pathogen infection with overall six patterns of TORCH infection and all infection patterns had no relationship with year and gender,respectively (P>0.05).Conclusion RV and CMV were primary pathogens in TORCH infection for neonate in Tianjin area and there were recent infections by CMV.TORCH infection varied in different years and gender groups,which provided experimental data and basis for epidemiology and prevention of TORCH in neonate.
10.Investigation on the Serum NGAL Reference Range of Healthy Populations in Xi'an
Ruijie SHI ; Futang YAN ; Wenkang LIU ; Hua ZHANG ; Jun YUAN ; Peng MI ; Shuling HU ; Xiao ZHENG
Journal of Modern Laboratory Medicine 2017;32(4):130-132
Objective To establish serum NGAL reference range of healthy populations in Xi'an Area.Methods 2 665 cases (aged 6 to 95 years old,male 1 370,female 1 295) of health-check people were collected from March 2014 to October 2016 in Medical Examination Center of Shaanxi Provincial People's Hospital,and 682 cases (aged 0 to 6 years old,male 356,female 326) were collected from preschool children of prevention.Serum NGAL concentration of them were analysed by immunoturbidimetry method with the Automatic Biochemical Analysis Assembly Line of Beckman-AU5800,and the detection data for statistical analysis.Then established the reference range of serum NGAL population of different age and different sex in Xi'an.Results The serum NGAL levels in healthy subjects showed a skewed distribution,which were preschool children under 6 years of age 37.66±23.12 ng/ml,6~15 years 39.25±25.34 ng/ml,16~49 years 46.68±27.06 ng/ml,and 50~ 69 years 57.82±29.13 ng/ml.Compared the first two with the latter,there was a significant difference (t=0.589,P< 0.05).The serum NGAL levels of over 70 years were 61.87 ± 32.64 ng/ml,and there was a significant difference between the ages of 15 and 49 and over 70 years (t=8.529,P<0.01).At the same time,the serum NGAL was closely correlated with age (r=0.298,P<0.01).But there was no significant difference in serum NGAL level between male and female (t=0.263~0.542,all P>0.05).87ng/ml was the upper limit of the reference value for the age of 50 years.Conclusion The level of serum NGAL was related to age and increased with age,but not with gender.

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