Objective:Study on the correlation between the active components of Salviea Miltiorrhizae Radix et Rhizoma screened by high-throughput sequencing and the regulation of lncRNA-mRNA in human lung adenocarcinoma A549 cells.Methods:A549 cells were cultured, and the IC 50 dose of cryptotanshinone and tanshinone ⅡA was confirmed according to the cell proliferation experiment. A549 cells were randomly divided into blank control group, cryptotanshinone group, and tanshinone IIA group using a random number table method. After 24 hours of intervention, the cell cycle was detected by flow cytometry. High-throughput sequencing technique was used to detect the expressions of lncRNA and mRNA in A549 cells in intervention group and non-intervention group. By analyzing the expression profiles of differential genes related to cryptotanshinone and tanshinone ⅡA, the obtained differential genes were analyzed by GO and KEGG. Results:The cell cycle results showed that the proportion of G0/G1 phase cells in cryptotanshinone and Tanshinone ⅡA was increased ( P<0.01), the proportion of S phase cells was decreased ( P<0.01), and the proportion of G2/M phase cells in cryptotanshinone was decreased ( P<0.01). The results of high-throughput screening showed that cryptotanshinone could up-regulate 4 698 lncRNA, down-regulate 1 557 lncRNA, up-regulate 4 810 mRNA and down-regulate 5 644 mRNA. Tanshinone ⅡA could up-regulate 1 348 lncRNA, down-regulate 1 299 lncRNA, up-regulate 4646 mRNA and down-regulate 4 741 mRNA. The function and pathway enrichment analysis of differential lncRNA and mRNA showed that the differentially expressed genes of cryptotanshinone and tanshinone ⅡA were mainly related to cell cycle, autophagy, AMPK signaling pathway, FoxO signaling pathway and EGFR signaling pathway. GAS5 may be one of the targets for the inhibitory effects of cryptotanshinone and tanshinone ⅡA. Conclusion:Cryptotanshinone and tanshinone ⅡA have certain inhibitory effects on A549 cells, and there are differentially expressed genes of lncRNA-mRNA, which are closely related to cell cycle and signal pathway.

Objective To analyze the relationships of carotid plaque characteristics and hemorheology with disease classification and severity in elderly patients with ischemic stroke. Methods A total of 240 elderly patients with ischemic stroke were selected as observation group, and were further divided into different groups based on disease classification [complete anterior circulation infarction (TACI), partial anterior circulation infarction (PACI), posterior circulation infarction (POCI), lacunar infarction (LACI)] and severity of the disease (mild, moderate, and severe); another 60 elderly healthy individuals with physical examinations were selected as control group. The characteristics of carotid artery plaques [carotid intima-media thickness (IMT), number of plaques, maximum length of plaques, and nature of plaques]as well as hemorheology indexes[erythrocyte sedimentation rate (ESR), hematocrit (HCT), plasma fibrinogen (Fib), plasma viscosity, and whole blood viscosity]were measured in all the cases, and the relationships of carotid plaque characteristics and hemorheology with disease classification and severity were analyzed. Results The IMT, the number of plaques, maximum diameter of plaques, ESR, HCT, Fib, plasma viscosity, and whole blood viscosity in the observation group were significantly higher than those in the control group (P < 0.05); the IMT, the number of plaques, and maximum diameter of plaques in the LACI group were significantly lower than those in the TACI, PACI, and POCI groups, while the ratios of stable plaques were significantly higher (P < 0.05); the ESR, HCT, Fib, plasma viscosity, and whole blood viscosity in the LACI group were significantly lower than those in the TACI, PACI, and POCI groups (P < 0.05); the IMT, the number of plaques, and maximum diameter of plaques in the mild group were significantly lower than those in the moderate and severe groups, the ratio of stable plaques was significantly higher than that in the moderate and severe groups, the IMT, the number of plaques, and maximum diameter of plaques in the moderate group were significantly lower than those in the severe group (P < 0.05); the ESR, HCT, Fib, plasma viscosity, and whole blood viscosity in the mild group were significantly lower than those in the moderate and severe groups, and the above indicators in the moderate group were significantly lower than those in the severe group (P < 0.05). Conclusion The characteristics of carotid artery plaques and hemorheology in elderly patients with ischemic stroke are related to the classification and severity of the disease, and the status of carotid artery plaques and hemorheological abnormalities in elderly patients with TACI, PACI, and POCI types of ischemic stroke are more severe than those of patients with LACI type.

Objective China is among the 30 countries with a high burden of tuberculosis(TB)worldwide,and TB remains a public health concern.Kashgar Prefecture in the southern Xinjiang Autonomous Region is considered as one of the highest TB burden regions in China.However,molecular epidemiological studies of Kashgar are lacking. Methods A population-based retrospective study was conducted using whole-genome sequencing(WGS)to determine the characteristics of drug resistance and the transmission patterns. Results A total of 1,668 isolates collected in 2020 were classified into lineages 2(46.0%),3(27.5%),and 4(26.5%).The drug resistance rates revealed by WGS showed that the top three drugs in terms of the resistance rate were isoniazid(7.4%,124/1,668),streptomycin(6.0%,100/1,668),and rifampicin(3.3%,55/1,668).The rate of rifampicin resistance was 1.8%(23/1,290)in the new cases and 9.4%(32/340)in the previously treated cases.Known resistance mutations were detected more frequently in lineage 2 strains than in lineage 3 or 4 strains,respectively:18.6%vs.8.7 or 9%,P<0.001.The estimated proportion of recent transmissions was 25.9%(432/1,668).Multivariate logistic analyses indicated that sex,age,occupation,lineage,and drug resistance were the risk factors for recent transmission.Despite the low rate of drug resistance,drug-resistant strains had a higher risk of recent transmission than the susceptible strains(adjusted odds ratio,1.414;95%CI,1.023-1.954;P = 0.036).Among all patients with drug-resistant tuberculosis(DR-TB),78.4%(171/218)were attributed to the transmission of DR-TB strains. Conclusion Our results suggest that drug-resistant strains are more transmissible than susceptible strains and that transmission is the major driving force of the current DR-TB epidemic in Kashgar.

Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.

Objective To explore the therapeutic effect and mechanism of pressing and rubbing method on myofascial pain syndrome(MPS)rats.Methods A total of 12 rats were randomly selected from 60 rats as the normal group,and the remaining rats were used to construct the MPS model by blunt strike combined with centrifugal exercise.Then 48 successfully modeled rats were randomly divided into model group,pressing and rubbing method group,pressing and rubbing method + Dantrolene[ryanodine receptor(RyR)inhibitor]group,pressing and rubbing method + normal saline group,with 12 rats in each group.The mechanical pain threshold was measured by von-Frey method.Detection of soft tissue tension,electromyography was performed;the ultrastructure of the pain point tissue was observed by transmission electron microscopy.The content of calcium ion(Ca2+)in the tissue of trigger point was detected by colorimetry.The protein expressions of dihydropyridine receptor(DHPR)α1,RyR and acetylcholinesterase(AChE)in the pain points of rats were detected by Western Blot.Results Compared with the normal group,the mechanical pain threshold,soft tissue tension in trigger point and the protein expressions of DHPRα1,RyR and AChE in the model group were decreased and the Ca2+ + content was increased(all P＜0.05),and the peak potential with higher amplitude was observed in the electromyogram.The ultrastructure of the trigger point tissue was damaged.Compared with the model group,the mechanical pain threshold,soft tissue tension of trigger point and the protein expressions of DHPRα1,RyR and AChE in the trigger point tissue of the rats in the pressing and rubbing method group and the pressing and rubbing method + normal saline group were increased,and the Ca2+ content was decreased(all P＜0.05),the electromyography was restored to be stable,the ultrastructural damage of the trigger point tissue was alleviated.Compared with the pressing and rubbing method group,the mechanical pain threshold,soft tissue tension of trigger point and protein the expressions of DHPRα1,RyR and AChE in the trigger point tissue of the rats in the pressing and rubbing method + Dantrolene group were decreased,and the Ca2+ content was increased(all P＜0.05),the electromyogram showed electrical activity changes,the ultrastructure of the trigger point tissue was damaged.Conclusion The pressing and rubbing method may improve MPS in rats by activating the DHPR/RyR signaling pathway.

Objective To observe the preventive effect of ultrasound-guided cervical vagus nerve block(CVB)in preventing oculocar-diac reflex.Methods A total of 86 patients who underwent posterior scleral reinforcement or ophthalmectomy were recruited and randomly divided into the general anesthesia group and the composite anesthesia group.After admission,patients in the two groups underwent conven-tional general anesthesia induction intubation,and the composite anesthesia group received another 1％lidocaine for right CVB under ultrasound guidance.The heart rate,blood pressure,saturation of peripheral oxygen,atropine usage and surgical procedures of the two groups were recorded.Postoperative comfort degree,hoarseness and complications related to nerve block were compared between the two groups.Results Compared with the general anesthesia group,the proportion of patients using atropine and the proportion of suspending operation in the composite anesthesia group were significantly reduced(P＜0.05),and the incidence of oculocardiac reflex and the level of oculocardiac reflex were significantly decreased(P＜0.05).The incidence of hoarseness in the composite anesthesia group was higher than that in the general anesthesia group,and the difference was statistically significant(P＜0.05).There was no complication related to nerve block occurred in the composite anesthesia group.Conclusion In ophthalmic surgery,ultrasound-guided right CVB can safely and effec-tively inhibit the occurrence of oculocardiac reflex.

This study established an efficient and specific method for type analysis of genetic variants of Balantioides coli.The restriction endonucleases ApoI and PflMI were selected to digest the PCR amplification products of ITS1-5.8S rDNA-ITS2 of B.coli,and to establish a PCR-RFLP typing method.The PCR-RFLP method was subsequently used to analyze the genetic variants in clinical fecal samples from pigs,sheep,and guinea pigs.The PCR-RFLP method based on ApoI and PflMI accurately distinguished the main A and B genetic variants of B.coli,and further divided the main B type into B-c and B-t sub-types of genetic variants with PflMI.Compared with the results of microscopy and sequencing,the PCR-RFLP method showed good specificity and higher sensitivity,and was able to identify not only single but also multiple variants of B.coli in a single clinical sample.This study successfully established the PCR-RFLP method for B.coli,which can be used for genetic di-versity identification and molecular epidemiological studies of B.coli.

Objective:To investigate the correlation of peripheral blood T lymphocyte subsets with overall survival(OS)and clinical baseline characteristics in mantle cell lymphoma(MCL).Methods:The clinical data of 55 MCL patients who were newly diagnosed in the Department of Hematology,Second Hospital of Shanxi Medical University from January 2012 to July 2022 were analyzed retrospectively.The percentages of T lymphocyte subsets and CD4+/CD8+ratio in peripheral blood were detected by flow cytometry,and their correlation with clinical characteristics of patients were analyzed.Kaplan-Meier method was used for survival analysis and survival curves were drawn.Log-rank test was used for univariate analysis,while Cox proportional hazards model was used for multivariate analysis.Results:The median follow-up was 40(1-68)months,and the median overall survival(OS)was 47 months.Among the 55 patients,30(54.5％)patients had a decrease in peripheral blood CD4+T lymphocyte,while 17(30.9％)patients had a increase in peripheral blood CD8+T lymphocyte,and 20(36.4％)patients had a decrease in CD4+/CD8+ratio.There were no significant correlations between CD4+/CD8+ratio and sex,age,Ki-67,B symptoms,leukocytes,hemoglobin,lymphocytes,platelets,albumin,lactate dehydrogenase(LDH),β2-microglobulin,splenomegaly,bone marrow invasion,primary site and MIPI score.Survival analysis showed that patients with CD4+T cell＞23.3％,CD8+Tcell ≤33.4％and CD4+/CD8+ratio＞0.6 had longer OS(P=0.020,P＜0.001,P＜0.001).Univariate analysis showed that Ki-67＞30％,LDH＞250 U/L,splenomegaly,bone marrow involvement,CD4+T cells 23.3％,CD8+T cells＞33.4％,CD4+/CD8+ratio ≤0.6 were adverse prognostic factors affecting OS of MCL patients.Multivariate analysis showed that CD4+/CD8+ratio ≤0.6(HR=4.382,P=0.005)was an independent adverse prognostic factor for OS of MCL patients.Conclusions:Low CD4+/CD8+ratio is associated with poor prognosis in MCL,and the CD4+/CD8+ratio can be used as an important indicator to evaluate the prognosis risk in MCL patients.

Objective:To investigate the expression of serum histone deacetylase 2(HDAC2)in patients with acute myeloid leukemia(AML)and its clinical significance.Methods:The 150 AML patients who received treatment in our hospital from April 2017 to April 2019 were included as AML group,and further divided into survival group(108 cases)and death group(42 cases)according to their survival status.In addition,100 health individuals undergoing health examination in the same period were included as control group.The expression of HDAC2 in serum was detected by real-time quantitative PCR.Cox regression was used to analyze the influencing factors of adverse prognosis.The predictive effect of serum HDAC2 for the adverse prognosis of AML patients was evaluated by the receiver operating characteristic(ROC)curve.Results:Compared with the control group,the expression of serum HDAC2,interleukin(IL)-6,tumor necrosis factor(TNF)-α,IL-1β and C-reactive protein(CRP)in AML group was obviously increased(all P＜0.05).Compared with the survival group,the expression of serum HDAC2 in the death group was also increased(P＜0.05).Serum HDAC2 was positively correlated with IL-6,TNF-α,IL-1β and CRP in AML patients(r=0.567,0.559,0.623,0.537).According to Cox regression analysis,the IL-6,TNF-α,IL-1 β and HDAC2 were independent risk factors affecting the prognosis of AML patients(all P＜0.05).ROC analysis showed that the AUC of serum HDAC2 level in predicting the adverse prognosis of AML was 0.862.Conclusion:The expression level of serum HDAC2 in AML patients is increased,which has positive correlations with IL-6,TNF-α,IL-1β and CRP.HDAC2 is an independent risk factor for the poor prognosis of AML patients,and has a high predictive value.

Objective To observe and compare the changes of vertebral bone mineral density(BMD)in the early stages of spinal Mycobacterium tuberculosis infection.Methods Patients who underwent spinal surgery at Xiangya Hospital,Central South University from January 1 to December 31,2023 were continuously enrolled(spinal tuber-culosis group),based on gender matching,non-spinal tuberculosis surgical patients treated for spinal stenosis were selected as the control group.Dual-energy X-ray scans were performed on the enrolled patients,difference in verte-bral BMD between two groups of patients was compared.An animal model of spinal Mycobacterium tuberculosis in-fection(referred to as the animal model)was constructed,differences in microstructure of trabecular bone between spinal tuberculosis group and control group was compared,and the bone volume/tissue volume(BV/TV),the thickness of trabecular bone(Tb.Th),the number of trabecular bone(Tb.N),and sparse density of trabecular(Tb.Sp)were used as evaluation indexes to further analyze the bone quality differences between the diseased verte-brae and the neighboring vertebrae.Results 69 patients were included in the spinal tuberculosis group and the con-trol group,respectively.The BMD of patients in the spinal tuberculosis group(0.793[0.712,0.869]g/cm2)was lower than that of the control group(0.907[0.800,1.020]g/cm2),difference was statistically significant(P＜0.05).Microstructure of trabecular bone BV/TV([18.4±5.4]％),Tb.Th([0.124±0.010]mm)in the spinal tuberculosis group of animal model were significantly altered compared with BV/TV([22.6±3.2]％),Tb.Th([0.160±0.017]mm)in the control group(both P＜0.05).In the spinal tuberculosis group,microstructure of diseased vetebral trabecular bone BV/TV([25.5±6.7]％)and Tb.N([1.871±0.443]/mm)were significantly lower than BV/TV([26.6±6.8]％)and Tb.N([1.969±0.454]/mm)in the neighboring vertebrae,both with statistically difference(both P＜0.05).Conclusion In the early stages of spinal Mycobacterium tuberculosis infec-tion,microstructure of vertebral trabecular bone can be altered,leading to a decrease in BMD.