Extracting extracts of secondary metabolites from the karst cave fungus Metarhizium anisopliae NHC-M3-2 from the Yilingyan Scenic Area in Guangxi. Ten compounds were isolated and purified from fungal secondary metabolites using thin-layer chromatography and high-performance liquid chromatography. 7-Hydroxy-3-hydroxypropyl-5,6-dimethylisochrome-1-one (1) and 7-hydroxy-3,5,6-trimethyl-isochromen-1-one (2) were new isocoumarin compounds, N-acetyl phenylalanine (3), chaetosumin J (4), 2-one-13-hydroxy-3,5,8,7(11)-eudesmatetraen-12,8-olide (5), 1H-indole-3-carboxaldehyde (6), irpexolaceus B (7), irlactin L (8), cytochalasin K (9), and helvolic acid (10), a total of 8 known compounds. The structure of the compound was determined using methods such as NMR and mass spectrometry. The tumor cytotoxicity of the compound was evaluated using the CCK-8 method. The results showed that the IC₅₀ of compound 2 on HepG2 cells was 29.83 µmol·L^-1, and compound 9 on HCT116 cells was 27.44 µmol·L^-1.

Objective To study the therapeutic value of Mongolian drug hatagaqi-7 for wound healing of diabetic ulcers in rats and preliminarily explore its molecular mechanism in regulating hypoxia-inducible factor-1α(HIF-1α).Methods Adult male SD rats were randomly divided into control,diabetes,Mongolian drug,and cytokine groups.Except in the control group,the other three groups were treated with an intraperitoneal injection of streptozotocin to establish the diabetes model.Ulcer wounds were prepared in the mouse back of the four groups.One week later,the Mongolian drug group was treated with hatagaqi-7,and the cytokine group was treated with recombinant bovine basic fibroblast growth factor for 2 consecutive weeks.Fasting blood glucose(FBG),wound area,wound pathology,expression levels of advanced glycation end products(AGEs),receptor of AGE(RAGE),HIF-1α and vascular endothelial growth factor(VEGF),secreted levels of interleukin-1β(IL-1β),interferon-γ(IFN-γ),and malondialdehyde(MDA),and the total antioxidant capacity(T-AOC)were assessed.Results FBG of diabetes,Mongolian drug and cytokine groups was higher than that in the control group(P<0.05),and no significant difference was observed among the three groups(P>0.05).Compared with the control group,the ulcer wound area,scope of unrepaired tissue,expression levels of AGEs and RAGE,and secreted levels of IL-1β,IFN-γ,and MDA in wound tissue of the diabetes group were increased,and T-AOC and expression levels of HIF-1α and VEGF of the diabetes group were decreased(P<0.05).Compared with the diabetes group,the ulcer wound area,scope of unrepaired tissue,expression levels of AGEs and RAGE,and secreted levels of IL-1β,IFN-γ,and MDA in wound tissue of Mongolian drug and cytokine groups were decreased,T-AOC and expression levels of HIF-1α and VEGF in Mongolian drug and cytokine groups were increased(P<0.05),and indexes of the Mongolian drug group were better than those of the cytokine group.Conclusions Mongolian drug hatagaqi-7 promotes ulcer wound healing in diabetic rats,the inhibiton of AGE and RAGE expression and induction of HIF-1 α are the possible molecular mechanism.

OBJECTIVE: In order to conduct high-throughput genome-wide translocation sequencing based on CRISPR/Cas9, Nalm6-cas9 monoclonal cell line expressing Cas9 protein was constructed by lentivirus transduction. METHODS: Lentiviral vectors LentiCas9-Blast, pSPAX2, and pMD2.G were used to co-transfect HEK293T cells to obtain recombinant lentivirus. After Nalm6 cells were infected with the recombinant lentivirus, the cells were screened by Blasticidin, and multiple monoclonal cell lines expressing Cas9 protein were obtained by limited dilution. Western blot was used to detect the expression level of Cas9 protein in monoclonal cell lines, and cell count analysis was used to detect the proliferation activity of monoclonal cell lines. LentiCRISPRV2GFP-Δcas9, LentiCRISPRV2GFP-Δcas9-AF4, LentiCRISPRV2GFP-Δ cas9-MLL plasmids were constructed, and transfected with pSPAX2 and pMD2.G, respectively. T vector cloning was used to detect the function of Cas9 protein in Nalm6-Cas9 monoclonal cell line infected with virus. RESULTS: Western blot showed that Nalm6-Cas9_1-6 monoclonal cell line had high expression of Cas9 protein. Cell count analysis showed that high expression of Cas9 protein in Nalm6-Cas9_1-6 monoclonal cell line did not affect cell proliferation activity. The Nalm6-Cas9_1-6 monoclonal cell line had high cleavage activity, and the editing efficiency of AF4 and MLL genes was more than 90% which was determined by T vector cloning. CONCLUSION Nalm6-Cas9_1-6 monoclonal cell line stably expressing highly active Cas9 protein was obtained, which provided a basis for exploring the translocation of MLL in therapy-related leukemias based on CRISPR/Cas9 genome-wide high-throughput genome-wide translocation sequencing.
CRISPR-Associated Protein 9/genetics* ; CRISPR-Cas Systems ; Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus/genetics* ; Plasmids

Objective: To compare the differences between CAS risk model and CHA₂DS₂-VASc risk score in predicting all cause death, thromboembolic events, major bleeding events and composite endpoint in patients with nonvalvular atrial fibrillation. Methods: This is a retrospective cohort study. From the China Atrial Fibrillation Registry cohort study, the patients with atrial fibrillation who were>18 years old were randomly divided into CAS risk score group and CHA₂DS₂-VASc risk score group respectively. According to the anticoagulant status at baseline and follow-up, patients in the 2 groups who complied with the scoring specifications for anticoagulation were selected for inclusion in this study. Baseline information such as age and gender in the two groups were collected and compared. Follow-up was performed periodically to collect information on anticoagulant therapy and endpoints. The endpoints were all-cause death, thromboembolism events and major bleeding, the composite endpoint events were all-cause death and thromboembolism events. The incidence of endpoints in CAS group and CHA₂DS₂-VASc group was analyzed, and multivariate Cox proportional risk model was used to analyze whether the incidence of the endpoints was statistically different between the two groups. Results: A total of 5 206 patients with AF were enrolled, average aged (63.6±12.2) years, and 2092 (40.2%) women. There were 2 447 cases (47.0%) in CAS risk score group and 2 759 cases (53.0%) in CHA₂DS₂-VASc risk score group. In the clinical baseline data of the two groups, the proportion of left ventricular ejection fraction<55%, non-paroxysmal atrial fibrillation, oral warfarin and HAS BLED score in the CAS group were lower than those in the CHA₂DS₂-VASc group, while the proportion of previous diabetes history and history of antiplatelet drugs in the CAS group was higher than that in the CHA₂DS₂-VASc group, and there was no statistical difference in other baseline data. Patients were followed up for (82.8±40.8) months. In CAS risk score group, 225(9.2%) had all-cause death, 186 (7.6%) had thromboembolic events, 81(3.3%) had major bleeding, and 368 (15.0%) had composite endpoint. In CHA₂DS₂-VASc risk score group, 261(9.5%) had all-cause death 209(7.6%) had thromboembolic events, 112(4.1%) had major bleeding, and 424 (15.4%) had composite endpoint. There were no significant differences in the occurrence of all-cause death, thromboembolic events, major bleeding and composite endpoint between anticoagulation in CAS risk score group and anticoagulation in CHA₂DS₂-VASc risk score group (log-rank P =0.643, 0.904, 0.126, 0.599, respectively). Compared with CAS risk score, multivariable Cox proportional hazards regression models showed no significant differences for all-cause death, thromboembolic events, major bleeding and composite endpoint between the two groups with HR(95%CI) 0.95(0.80-1.14), 1.00(0.82-1.22), 0.83(0.62-1.10), 0.96(0.84-1.11), respectively. All P>0.05. Conclusions: There were no significant differences between CAS risk model and CHA₂DS₂-VASc risk score in predicting all-cause death, thromboembolic events, and major bleeding events in Chinese patients with non-valvular atrial fibrillation.
Adolescent ; Anticoagulants ; Atrial Fibrillation/drug therapy* ; Cohort Studies ; Female ; Hemorrhage/complications* ; Humans ; Male ; Retrospective Studies ; Risk Assessment ; Stroke/epidemiology* ; Stroke Volume ; Thromboembolism/etiology* ; Ventricular Function, Left

Background:Transcranial alternating current stimulation (tACS) offers a new approach for adult patients with major depressive disorder (MDD).The study is to evaluate the efficacy and safety of tACS treating MDD.Methods:This is an 8-week,double-blind,randomized,placebo-controlled study.Ninety-two drug-naive patients with MDD aged 18 to 65 years will receive 20 daily 40-min,77.5-Hz,15-mA sessions of active or sham tACS targeting the forehead and both mastoid areas on weekdays for 4 consecutive weeks (week 4),following a 4-week observation period (week 8).The primary outcome is the remission rate defined as the 17-item Hamilton depression rating scale (HDRS-17) score ≤7 at week 8.Secondary outcomes are the rates of response at weeks 4 and 8 and rate of remission at week 4 based on HDRS-17,the proportion of participants having improvement in the clinical global impression-improvement,the change in HDRS-17 score (range,0-52,with higher scores indicating more depression) over the study,and variations of brain imaging and neurocognition from baseline to week 4.Safety will be assessed by vital signs at weeks 4 and 8,and adverse events will be collected during the entire study.Discussion:The tACS applied in this trial may have treatment effects on MDD with minimal side effects.

There are many problems in field training of mobile medical service units, such as lack of training ground, backward training methods, simple training methods and weak comprehensive treatment ability. The purpose of this study is to explore the research and development idea and application effect of mobile combat-injury treatment training assessment system, which consists of handheld mobile terminals, 3 function modules for training mode, training evaluation and inquiry, and 5 basic modules for comprehensive appraisal of training performance, recording and analysis of training results, facilitation of personalized training programs, multi-view summary of training performances and intelligent stratified management of trainees, respectively. This mobile assessment system makes the training equipment more portable, training means more convenient, training activities derestrict to time, place or weather and, as a result, the whole combat-injury treatment training more efficient.

Based on rDNA ITS sequences of Dendrobium officinale and the other 69 species of Dendrobium, a pair of dismatched allele-specific diagnostic primers, TPSH-AS1F and TPSH-AS1R were designed to authenticate D. officinale from the other species. Thebidirectional PCR amplification were performed using the diagnostic primers with the total DNAs of the original plants or processing products as a template. When the annealing temperature was raised to 60 ℃, only the template DNA of D. officinale could be amplified whereas the diagnostic PCRs of the other Dendrobium species were all negative. Compared with the other authentification methods, the bidirectional PCR amplifications is not only simpler and time-saving but practical and effective.

Objective To investigate the endothelial dysfunction in pre-hypertension and its influencing factors.Methods A total of 373 youth were divided as the subjects into hypertension group (HBP group),prehypertension group (PHT group) and normal blood pressure group (NBP group).Endothelial function was assessed based on carotid intima-media thickness (IMT),brachial artery flow-mediated dilation (FMD) and brachial-ankle pulse wave velocity (baPWV).Results IMT and baPWV in PHT group were higher than those in NBP group (P＜0.05),but did not reach the significant difference when compared with HBP group (P＞0.05).Compared with HBP,the levels of FMD in PHT group significantly increased (P＜ 0.05);however,no difference was observed in comparison with NBP group (P＞0.05).In the early stage of hypertension,diastolic BP (β=-0.120,P＜0.05) and body mass index (β=-0.115,P＜0.05) were negatively correlated with FMD;diastolic BP (β=0.146,P＜0.05),2-hour glucose (β=0.147,P＜0.05),high-density lipoprotein cholestrol (β=0.150,P＜0.05),and waist-hip ratio (β=0.126,P＜0.05) showed a positive correlation with IMT.baPWV was correlated with systolicBP (β=0.358,P＜0.01),waist circumference (β=0.254,P＜0.05),fasting glucose (β=0.155,P＜0.05),postprandial 2 h blood glucose (β =0.152,P ＜0.05),uric acid (β =0.206,P ＜ 0.05),and C-reactive protein (β=0.099,P＜0.05).Corclusion Our study shows that endothelial dysfunction may exist in the prehypertensive young,and several cardiovascular risks contribute to its development in the early stage of hypertension.

In order to evaluate the performance of a molecular Hain line probe assay (Hain LPA) for rapid detection of rifampicin and isoniazid resistance of Mycobacterium tuberculosis in China, 1612 smear positive patients were consecutively enrolled in this study. Smear positive sputum specimens were collected for Hain LPA and conventional drug susceptibility testing (DST). The sensitivity and specificity of Hain LPA were analyzed by using conventional DST as golden reference. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for rifampicin resistance detection were 88.33%, 97.66%, 81.54%, and 98.62%, respectively. The sensitivity, specificity, PPV and NPV for isoniazid resistance detection were 80.25%, 98.07%, 87.25%, and 96.78%, respectively. These findings suggested that Hain LPA can be an effective method worthy of broader use in China.
China ; Genotyping Techniques ; methods ; Humans ; Isoniazid ; pharmacology ; Mycobacterium tuberculosis ; drug effects ; genetics ; isolation & purification ; Rifampin ; pharmacology ; Tuberculosis, Multidrug-Resistant ; diagnosis ; microbiology

OBJECTIVETo investigate the association of childhood hemophagocytic syndrome (HPS) with human parvovirus B19 (HPVB19) infection, and to analyze the clinical features of this disease.

METHODSELISA and quantitative real-time PCR were used to detect HPVB19-IgM, HPVB19-IgG and HPVB19-DNA in 65 children with HPS (HPS group) and 65 healthy children (control group). The HPS group was divided into HPVB19-infected (n=14) and non-infected (n=51) groups according to the detection results of HPVB19-DNA. The clinical data of two groups were compared.

RESULTSThe positive rate of HPVB19-IgM in the HPS group (26%, 17/65) was significantly higher than that in the control group (9%, 6/65) (P=0.011), and there was no significant difference in the positive rate of HPVB19-IgG between the HPS (38%, 25/65) and control groups (29%, 19/65) (P=0.266). The infection rate of HPVB19 in the HPS group (22%, 14/65) was significantly higher than that in the control group (3%, 2/65) (P=0.001). Compared with the non-infected group, the HPVB19-infected group had significantly lower platelet count and hemoglobin level on admission, significantly more severe liver function damage, a significantly earlier onset time, and a significantly longer course of disease (P<0.05).

CONCLUSIONSThe pathogenesis of HPS may be associated with HPVBl9 infection. HPVBl9-infected children with HPS have more acute onset, more severe clinical manifestations, and a longer disease duration.

Adolescent ; Antibodies, Viral ; analysis ; Child ; Child, Preschool ; DNA, Viral ; analysis ; Female ; Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; etiology ; Male ; Parvoviridae Infections ; complications ; Parvovirus B19, Human