Objective To characterize the temporal dynamics of endophytic community and key metabolites during the growth developmental stages of Artemisia indica and to explore the effects of endophytes on the medicinal material quality of A.indica from the perspective of microorganism combined with the correlation analysis.Methods In this study,high-throughput sequencing was performed to obtain the temporal dynamics of diversity and composition of endophytic community during the growth stages of A.indica.Key metabolites quantitative analysis were performed using high-performance liquid chromatography and ultraviolet spectrophotometry.Spearman's correlation analysis was further conducted to identify the endophytic communities that were significantly correlated with metabolites.Results There were significant differences in the diversity and composition of endophytic communities during the growth stages of A.indica.Linear discriminant analysis efect size(LEfSe)analyses identified some potential microbial biomarkers,such as Firmicutes,Massilia,Pantoea,Alternaria,Didymella,and Trichomerium.The abundances of some genera were significantly and positively correlated with the content of total polyphenols and total flavonoids(P<0.05),such as Curtobacterium,Pantoea,Tilletiopsis,and Dissoconium.The abundances of Pseudozyma,Ralstonia,and Pleospora were significantly and positively correlated with Chlorogenic acid,Isochlorogenic acid B,Isochlorogenic acid A,and Isochlorogenic acid C(P<0.05).Conclusion This work described the distribution of endophytes and key metabolites during the development stages of A.indica.Some endophytic communities with significant correlation of key metabolites were identified,which provided valuable information to guide the isolation of endophytic strains related to key metabolites and improve the quality of A.indica.

Objective To establish a HPLC method and fingerprints study for the simultaneous determination of loganic acid,gentiopicroside,paeoniflorin,vitexin,liquiritin,and ammonium glycyrrhizinatein,and other components in the alcohol extract of Mongolian medicine Qinggan Manu-4.Methods The Agilent Eclipse XDB-C18 column(250 mm×4.6 mm,5 μm)was used,with the detection wavelength of 230 nm for paeoniflorin,vitexin and liquiritin,237 nm for loganic acid,250 nm for glycyrrhizic acid,and 270 nm for gentiopicroside.The mobile phase was 0.1%phosphoric acid aqueous solution-acetonitrile with gradient elution.The flow rate was 1.0 mL/min,the column temperature was 30℃,and the injection volume was 10 μL.Results The mass concentrations of loganic acid,gentiopicroside,paeoniflorin,vitexin,liquiritin and ammonium gentiopicroside in the alcohol extract of Mongolian medicine Qinggan Manu-4 showed good linear relationships with the chromatographic peak area in the range of 0.020 3-0.121 0,0.053 3-0.317 7,0.021 3-0.127 0,0.011 5-0.069 0,0.014 1-0.083 8 and 0.035 1-0.209 1 mg/mL,respectively(r≥0.999 8).The average recovery rates of the six components were 94.75%,99.49%,92.32%,95.82%,101.29%and 98.04%,with the RSDs of 1.11%,0.76%,0.99%,2.75%,1.09%and 2.43%,respectively(n=6).The HPLC fingerprint of the alcoholic extract of Mongolian medicine Qinggan Manu-4 was established,using the chromatogram of sample S1 as the reference chromatogram,the control fingerprints were generated through multi-point correction and full-spectrum peak matching.A total of ten common peaks were identified,and after comparison with the reference substance,eight components were identified.Conclusion The established method and fingerprints are accurate,reliable,reproducible and specific,which provide a basis for the quality control and subsequent development of Mongolian medicine Qinggan Manu-4.

Objective:To retrospectively analyze the treatment status of tyrosine kinase inhibitors (TKI) in newly diagnosed patients with chronic myeloid leukemia (CML) in China.Methods:Data of chronic phase (CP) and accelerated phase (AP) CML patients diagnosed from January 2006 to December 2022 from 77 centers, ≥18 years old, and receiving initial imatinib, nilotinib, dasatinib or flumatinib-therapy within 6 months after diagnosis in China with complete data were retrospectively interrogated. The choice of initial TKI, current TKI medications, treatment switch and reasons, treatment responses and outcomes as well as the variables associated with them were analyzed.Results:6 893 patients in CP ( n=6 453, 93.6%) or AP ( n=440, 6.4%) receiving initial imatinib ( n=4 906, 71.2%), nilotinib ( n=1 157, 16.8%), dasatinib ( n=298, 4.3%) or flumatinib ( n=532, 7.2%) -therapy. With the median follow-up of 43 ( IQR 22-75) months, 1 581 (22.9%) patients switched TKI due to resistance ( n=1 055, 15.3%), intolerance ( n=248, 3.6%), pursuit of better efficacy ( n=168, 2.4%), economic or other reasons ( n=110, 1.6%). The frequency of switching TKI in AP patients was significantly-higher than that in CP patients (44.1% vs 21.5%, P<0.001), and more AP patients switched TKI due to resistance than CP patients (75.3% vs 66.1%, P=0.011). Multi-variable analyses showed that male, lower HGB concentration and ELTS intermediate/high-risk cohort were associated with lower cytogenetic and molecular responses rate and poor outcomes in CP patients; higher WBC count and initial the second-generation TKI treatment, the higher response rates; Ph + ACA at diagnosis, poor PFS. However, Sokal intermediate/high-risk cohort was only significantly-associated with lower CCyR and MMR rates and the poor PFS. Lower HGB concentration and larger spleen size were significantly-associated with the lower cytogenetic and molecular response rates in AP patients; initial the second-generation TKI treatment, the higher treatment response rates; lower PLT count, higher blasts and Ph + ACA, poorer TFS; Ph + ACA, poorer OS. Conclusion:At present, the vast majority of newly-diagnosed CML-CP or AP patients could benefit from TKI treatment in the long term with the good treatment responses and survival outcomes.

Objective:To investigate the influence of hepatitis B virus (HBV) combined with human immunodeficiency virus (HIV) infection on the efficacy of anti-retroviral therapy (ART).Methods:The data of 269 HIV-infected patients treated in Chongqing Public Health Medical Center from September 2016 to October 2019 were collected. The patients were divided into HIV monoinfection group and HIV/HBV coinfection group. The changes in liver function, CD4 + T lymphocyte count, and HIV RNA level between the two groups were compared when ART started and at different time points (2, 4, 8, 12, 24, 36, 48, and 96 weeks) after ART started. Statistical analysis were performed by independent sample t test, rank sum test and chi-square test. Results:A total of 145 patients with HIV monoinfection and 124 patients with HIV/HBV coinfection were collected. There were no statistically significant differences in liver function indexes (aspartate aminotransferase ( t=9.566), alanine aminotransferase ( t=-4.652) and total bilirubin ( t=-25.476)) between the two groups of patients when ART started (all P>0.05). At 24, 48 and 96 weeks after ART, the CD4 + T lymphocyte counts in the HIV monoinfection group and the HIV/HBV coinfection group were (305.9±156.9)/μL vs (266.2±172.5)/μL, (388.5±226.1)/μL vs (380.8±287.4)/μL and (369.5±191.4)/μL vs (453.6±179.6)/μL, respectively. At 48, 72 and 96 weeks after ART, the CD4 + T lymphocyte count increasing values were 121.0(-52.5, 144.5)/μL vs 156.0(-35.8, 185.8)/μL, 139.0(-116.0, 176.8)/μL vs 114.5(-59.5, 229.0)/μL and -91.0(-110.0, 153.3)/μL vs -94.0(-130.8, 114.3)/μL, respectively. The differences were all not statistically significant ( t=-0.516, -0.066 and -1.414, Z=-1.715、-0.802 and -1.602, respectively, all P>0.05). At 24, 48, and 96 weeks after ART, the HIV RNA inhibition rates in the HIV monoinfection group were 89.7%(130/145), 96.6%(140/145), and 96.6%(140/145), respectively, and those in the HIV/HBV coinfection group were 87.1%(108/124), 92.7%(115/124) and 94.4%(117/124), respectively. The differences were all not statistically significant ( χ2=0.026, 0.053 and 0.017, respectively, all P>0.05). In the second and fourth weeks after ART, the abnormal liver function rates of the HIV monoinfection group were 3.4%(5/145) and 6.2%(9/145), respectively, which were lower than those in the HIV/HBV coinfection group (21.0%(26/124) and 13.7%(17/124), respectively). The differences were both statistically significant ( χ2=20.121 and 4.309, respectively, both P<0.05). However, the abnormal liver function rates in the two group in the 8th week after ART were 10.3%(15/145) and 9.7%(12/124), respectively, and those in the 12th week were 9.0%(13/145) and 9.7%(12/124), respectively, and those in the 24th week were 9.7%(14/145) and 8.9%(11/124), respectively, and those in the 36th week were 9.7%(14/145) and 10.5%(13/124), respectively, and those in the 48th week were 8.3%(12/145) and 8.1%(10/124), respectively, and those in the 96th week were 2.8%(4/145) and 0(0/124), respectively. The differences were all not statistically significant ( χ2=0.330, 0.040, 0.049, 0.051, 0.004 and 3.472, respectively, all P>0.05). Conclusion:HBV coinfection has no adverse effect on the ART effect of HIV-infected patients.

Objective:To investigate the relationship between calf circumference and appendicular skeletal muscle mass (ASM) index, handgrip strength and gait speed in the elderly in Shanghai, and to explore the optimal cut-off point of calf circumference as a simple proxy marker of skeletal muscle mass, strength and function.Methods:A total of 2 294 participants were recruited from physical examination centers in Huadong Hospital Affiliated to Fudan University and 4 communities (954 males and 1 340 females) with an average age of (71.6±8.7) years. Appendicular skeletal muscle mass (ASM), Upper (UMM) and lower (LMM) limbs skeletal muscle mass were measured by bioelectrical impedance analysis (BIA). The ASM index (ASM/height 2) were calculated.Gait speed was measured.Muscle strength was measured by handgrip strength using an electronic hand dynamometer.Height, weight, right calf circumference and waist circumference were measured. Results:The age, body mass index (BMI), waist circumference and gait speed were not statistically different (all P>0.05) between both genders. Height, weight, ASM, ASM/height 2, UMM, LMM, handgrip strength, calf circumference were significantly higher in men than in women (all P<0.05). The calf circumference of the elderly with age ≥ 80 was significantly lower than that of the 70-79 age group and 60-69 age group ( P<0.05). The calf circumference of older women (age≥80) was significantly lower than that of the elderly women in the 70-79 age group and 60-69 age group ( P<0.05). Calf circumference was correlated negatively with age( P<0.01) and positively with ASM/height 2, height, weight, BMI, handgrip strength, UMM and LMM( P<0.01). There was no correlation between calf circumference and gait speed ( P>0.05). In the receiver operating characteristic curve (ROC), the cut-off value of calf circumference for low muscle mass was 35.1 cm for male(specificity, 0.721; sensitivity, 0.803, ROC, 0.809) and 33.6 cm for female(specificity, 0.774; sensitivity, 0.778; ROC, 0.827). Conclusions:Calf circumference decreases with ageing in the elderly. The optimal cut-off value of calf circumference for low muscle mass is 35.1 cm for males and 33.6 cm for females. We should pay attention to the importance of calf circumference in evaluation and diagnosis of sarcopenia.

Objective:To evaluate the role of connexin43 (Cx43) in sevoflurane anesthesia-induced cognitive dysfunction in aged rats.Methods:Fifty-two healthy male Sprague-Dawley rats, aged 18 months, weighing 400-500 g, were divided into 4 groups ( n=13 each) using a random number table method: control group (C group), sevoflurane group (SEV group), sevoflurane plus sh-NC group (SEV+ sh-NC group) and sevoflurane plus sh-Cx43 group (SEV+ sh-Cx43 group). Sevoflurane anesthesia model was established by inhaling 3% sevoflurane for 6 h. In SEV+ sh-NC group and SEV+ sh-CX43 group, sh-NC 5 nmol and sh-CX43 5 nmol were transfected into the lateral ventricles, respectively, at 1 day before sevoflurane anesthesia.Morris water maze test was performed at 30 min before anesthesia and 1, 2 and 3 days after the end of anesthesia, and the rats were sacrificed at each time point after Morris water maze test, the brains were removed, and the hippocampi were isolated for microscopic examination of the pathological changes and for determination of the expression of Cx43, cleaved caspase-3 and cleaved caspase-9 (by using Western blot), and contents of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and IL-6 (by enzyme-linked immunosorbent assay). Results:Compared with group C, the escape latency was significantly prolonged, and the number of crossing the original platform was reduced, and the expression of CX43, cleaved caspase-3 and cleaved caspase-9 was up-regulated, and the contents of IL-1β, TNF-α and IL-6 were increased in group SEV ( P<0.05). Compared with group SEV, the escape latency was significantly shortened, the number of crossing the original platform was increased, and the expression of CX43, cleaved caspase-3 and cleaved caspase-9 was down-regulated, the contents of IL-1β, TNF-α and IL-6 were decreased ( P<0.05), and the hippocampal pathological injury was reduced in group SEV+ sh-CX43, and no significant change was found in the indicators mentioned above in group SEV+ sh-NC ( P>0.05). Conclusion:Cx43 is involved in the pathophysiological mechanism of sevoflurane-induced cognitive dysfunction probably by inducing neuroinflammatory responses and cell apoptosis in aged rats.

Objective To investigate the relationship between calf circumference and skeletal muscle mass, strength and function in the elderly in Shanghai. Methods A total of 401 old people were recruited from physical examination centers in our hospital with the mean age of (71. 8±7. 4) years, including 202 males and 199 females. Adipose tissue, skeletal muscle mass (SMM) and appendicular skeletal muscle mass (ASM) were measured by bioelectrical impedance analysis. The ASM index ( ASM/height2) and SMM index ( SMM/height2) were calculated. Muscle function was measured by testing gait speed (m/s). Muscle strength was measured by handgrip strength (HGS) using an electronic hand dynamometer, with the dominant hand gripping for three times and taking the maximum value. Height, weight, right calf circumference and waist circumference were measured. Results Height, weight, ASM, ASM/height2, SMM, SMM/height2and HGS were significantly higher in men than in women ( P<0. 01); adipose tissue was significantly higher in women than in men (P<0. 05). The age, body mass index, calf circumference, waist circumference and gait speed were not statistically different (P=0. 426, P=0. 143, P=0. 425, P=0. 093, P=0. 143). There were linear correlations between calf circumference and height, weight, BMI, waist circumference, ASM, ASM/height2, adipose tissue, grip strength, SMM and SMM/height2( P=0. 001, P<0. 001, P<0. 001, P<0. 001, P<0. 001, P<0. 001, P<0. 001, P=0. 003, P<0. 001, P<0. 001). There was no linear correlation between calf circumference and pace and age (P=0. 818, P=0. 078). Multiple stepwise regression analysis showed that height, BMI, ASM/height2and SMM were correlated with the calf circumference. Conclusions Skeletal muscle mass and strength are higher in men than in women, while adipose tissue is higher in women than in men. Skeletal musle mass is an important factor affecting the calf circumference. There is a linear correlation be-tween skeletal muscle strength and calf circumference.

Objective To explore the features of coronary artery morphological changes and left ventricular function indicators of resident population in plateau .Methods 50 plateau healthy adults and 50 plain‐and Dalian‐healthy volunteers completed coronary CTA examination .We measured the pipe diameter of left main coronary atery and right coronary artery and their branches ,respec‐tively .Furthermore ,SV and EF value of left ventricular cardiac function indexes and the wall thickening of free ventricular wall at left ventricular end systolic were measured .Results The average diameter of proximal right coronary of plain‐and Dalian‐healthy volun‐teers thicker than plateau group ,with statistical difference ;Other coronary artery branch did not exhibit any significant between‐group differences ;The two groups did not show significant between‐group differences with heart function indexes .Conclusion The body circulation system have adaptability and compensatory changes of resident population in plateau ,and their coronary artery mor‐phology and left ventricular function are as similar as plain people .

Objective To investigate the effect of curcumin (Cur)on AngⅡ-induced proliferation and oxidative stress of vascular smooth muscle cells (VSMCs).Methods Primary rat VSMCs were cultured and divided into control group,AngⅡ group,AngⅡ+Cur 5μmol/L group,AngⅡ+Cur 10μmol/L group,AngⅡ+Cur 20μmol/L group,and Cur 20μmol/L group.The proliferation of AngⅡ-induced VSMCs was measured by MTT assay.The mRNA and protein expressions of inducible nitric oxide synthase (iNOS)and p47phox were detected by real-time PCR and Western blot.Nitric oxide (NO)production was measured by Griess reaction.Production of intracellular reactive oxygen species (ROS)was measured by DCFH-DA staining,and the activities of superoxide dismutase (SOD)and glutathione peroxidase (Gpx)were detected by xanthine oxidase assay and visible spectrophotometer. small interfering RNA (siRNA)was used to silence the expression of p47phox to further explore the mechanism for Cur inhibiting the proliferation of AngⅡ-induced VSMCs and oxidative stress.Results VSMCs activities were not significantly affected by Cur at the concentration between 0 and 80μmol/L.Cur (5,10 and 20μmol/L)significantly inhibited AngⅡ-induced proliferation of VSMCs.Cur had an inhibitory effect on the overexpression of NO,iNOS, p47phox and ROS in VSMCs and upregulated the activities of SOD and Gpx in a concentration-dependent manner. AngⅡ-induced ROS production in VSMCs was significantly attenuated by pretreatment with p47phox specific siRNA.Conclusion Cur can inhibit the proliferation and oxidative stress of AngⅡ-induced VSMCs.

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