Objective To investigate the diagnostic value of Xpert MTB/RIF combined with metagenic high-throughput sequencing in bacterial negative tuberculosis.Methods Retrospective analysis of 70 patients diagnosed with bacterial negative pulmonary tuberculosis from December 2019 to May 2022 enrolled in the study.Xpert MTB/RIF testing,high-throughput metagenomic sequencing,and a combination of the two were performed,respectively.Solid culture and proportional methods were used to diagnose biochemical pathogens and analyze diagnostic efficacy.The receiver operating characteristic curve was drawn with the predicted value.Results The positive rate of Xpert MTB/RIF diagnosis was 75.57%,and the negative rate was 21.43%.The positive rate of high-throughput sequencing for metagenomics was 78.57%,while the negative rate was 21.43%;The positive rate of combined diagnosis was 88.57%,and the negative rate was 11.43%.The detection sensitivity of Xpert MTB/RIF was 77.55%,the specificity was 59.86%,and the area under the curve(AUC)was 0.827.The sensitivity of high-throughput sequencing for metagenomes was 77.47%,the specificity was 61.02%,and the AUC was 0.808.The sensitivity of the combined detection was 89.75%,the specificity was 89.57%,and the AUC was 0.925.The results showed that the diagnostic sensitivity and specificity of Xpert MTB/RIF combined with high-throughput metagenomic sequencing in bacterial negative pulmonary tuberculosis were higher than those of single detection(P<0.05).Conclusion Xpert MTB/RIF combined with metagenic high-throughput sequencing has good application value in the diagnosis of bacterial negative pulmonary tuberculosis,and is worthy of clinical application.

Objective To optimize the flash extraction process of Huangyu Shenshu Granules based on Box-Behnken design-response surface methodology. Methods The extraction yields of morroniside and loganin, as well as the yield of dry extract, were used as evaluation indices. The rotation speed, liquid-solid ratio, and extraction time were selected as the investigation factors. Based on preliminary experiments, combined with Box-Behnken design-response surface methodology, the flash extraction process of Huangyu Shenshu Granules was studied and optimized. Results The optimal condition for the flash extraction of Huangyu Shenshu Granules were taking water as the extraction solvent, with a rotation speed of 4, 560 r/min, a liquid-solid ratio of 23 mL/g, and an extraction time of 85 s. Conclusion The flash extraction process established based on Box-Behnken design-response surface methodology is rapid and efficient, and suitable for the subsequent industrialization development of Huangyu Shenshu Granules.

Leptomeningeal metastasis(LM)is a serious late complication in patients with solid tumors,which is common in patients with lung cancer,breast cancer and melanoma.In recent years,due to the progress of diagnosis,the diagnosis rate of LM has gradually increased.The main goal of the therapy is to maintain neurological function,improve the quality of life and the overall survival rate,and prolong the progression-free survival time of patients.Intrathecal therapy is one of the main treatments for LM,which can deliver drugs directly to the subarachnoid space.The traditional intrathecal drugs are methotrexate,cytarabine and thiotepa.With the development of new research,a variety of chemotherapeutic drugs,targeted drugs and immune checkpoint inhibitors have also been used in intrathecal therapy.In addition,different ways of intrathecal administration also bring new hope to patients.This article reviewed the clinical research progress of intrathecal therapy in the treatment of LM.

Objective:To investigate the effects of compound Duzhong Jiangu Granules on joint function and gut microbiota in patients with Kashin-Beck disease.Methods:A single group pre- and post-experimental design was conducted, the patients with Kashin-Beck disease were selected as the subjects in Xunyi County, Xianyang City, Shaanxi Province; and treated with oral administration of compound Duzhong Jiangu Granules (12 g/bag, 1 bag/time, 3 times/day) for a period of 1 month. The improvement of joint function was evaluated using the joint dysfunction index scoring method before and after treatment. Morning stool samples of patients were collected and the changes in gut microbiota were analyzed before and after treatment using 16S rDNA sequencing technology.Results:A total of 87 patients with Kashin-Beck disease were included, including 44 males and 43 females; the age was (60.38 ± 7.12) years old, and the body mass index was (23.67 ± 3.59) kg/m 2. The comprehensive scores of joint dysfunction index for patients with Kashin-Beck disease before and after treatment were (7.27 ± 2.05) and (5.86 ± 2.01) points, respectively, and the difference was statistically significant ( t = 5.88, P < 0.001). The sequencing results of gut microbiota showed that there were statistically significant differences in the alpha diversity (chao1, observed species index) and beta diversity of gut microbiota in patients with Kashin-Beck disease before and after treatment ( Z = - 5.08, - 5.03, R = 0.09, P < 0.001). In the distribution of gut microbiota, Firmicutes was the dominant phylum, with relative abundances of 50.21% and 52.09% before and after treatment, respectively; the Bifidobacterium was the dominant bacterial genus, with relative abundances of 16.83% and 18.81% before and after treatment, respectively. At the genus level, a total of 17 gut microbiota genera were screened out, among which the relative abundances of Hafnia-Obesumbacterium, Gammaproteobacteria_unclassified, Acinetobacter, Pantoea, Leuconostoc, and Akkermanisia were significantly higher than before treatment ( Z = - 2.40, - 2.24, - 2.06, - 3.59, - 2.24, - 2.11, P < 0.05). The relative abundances of Dubosiella, Selenomonas, Anaeroplasma, Lachnospiraceae_ NK4A136_group, Rikenella, Prevotella, Megasphaera, Lactobacillus, Prevotella-9, Phascolarctobacterium, and Desulfovibrio were significantly lower than before treatment ( Z = - 9.38, - 2.61, - 2.18, - 8.43, - 2.45, - 2.46, - 2.49, - 7.29, - 2.29, - 2.55, - 2.08, P < 0.05). Conclusions:Compound Duzhong Jiangu Granules can effectively improve the joint function of patients with Kashin-Beck disease, and alter the diversity and richness of the gut microbiota community. It may reduce clinical symptoms in patients by regulating the structure of gut microbiota.

The exploration of anesthesia and consciousness has always been an important subject in neuroscience,but the underlying neural mechanisms of consciousness are still unclear,which limits the de-velopment of anesthesia monitoring and consciousness evaluation systems.The neural correlates of conscious-ness(NCC)cannot be determined by a single brain region or mechanism,suggesting that consciousness may arise from complex interactions of brain functions on space and time scales.In order to characterize these interactions,network science has been introduced in exploring the mechanisms of consciousness.In this paper,the basic concepts and common indicators of brain network research are introduced,and the lat-est progress of brain network research in anesthesia is reviewed,so as to provide new ideas for the applica-tion of brain network indicators in clinical monitoring and provide directions for the exploration of NCC.

Objective:To explore the application value of histogram and texture feature analysis based on CT images in distinguishing long bone osteosarcoma (OS) from Ewing sarcoma (ES).Methods:A retrospective collection of 25 patients with long bone osteosarcoma and 25 patients with Ewing sarcoma confirmed by surgery and pathology in National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Qilu Hospital of Shandong University and Nanjing Drum Tower Hospital, Nanjing University Medical School, from March 2018 to May 2023 was conducted. All patients were randomly divided into a training set (21 cases of OS and 19 cases of ES) and a validation set (4 cases of OS and 6 cases of ES) in an 8∶2 ratio. The region of interest (ROI) on CT images to extract texture feature parameters was manually sketched. Random forest and least absolute shrinkage and selection operator (LASSO) algorithm were used for feature screening. Logistic regression (LR), random forest (RF), support vector machine (SVM) and K-nearest neighbor (KNN) classifiers were used to establish models respectively. Receiver operating characteristic (ROC)curve was drawn and area under the curve (AUC) was calculated to evaluate the diagnostic efficiency of the four models.Results:A total of 100 texture parameters were extracted from CT images, and 8 feature parameters (maximum 3D diameter, 10th percentile, kurtosis, maximum pixel intensity value, inverse normalization, grayscale level variance, long range high grayscale emphasis, and low grayscale area emphasis) were obtained through screening. Four classifiers were used to establish models, and the AUC values of the four models (LR, RF, SVM, KNN) in the validation group were 0.92, 0.79, 0.83, and 0.73, respectively. LR and SVM classifier algorithm trains models had high diagnostic efficiency, with an accuracy of 90%, sensitivity of 83%, specificity of 100%, and AUC of 92% for the LR classifier validation set; the accuracy of SVM classifier validation set was 80%, sensitivity was 67%, specificity was 100%, and AUC was 83%.Conclusions:LR and SVM models have high value in distinguishing OS and ES.

In the earliest days,the idea that surviving a single infection often resulted in lifelong immunity to the infecting pathogen was recorded and then led to the discovery of vaccination.We have now confirmed that such protection is primarily based on the generation of immunological memory in antibody response.With the wide implementation of more and more vaccines around the world,it is well documented that different vaccines have different potential regarding to the duration of antibody response.In clinical observations,live-attenuated vaccines often elicit long-term immunity but are also accompanied with risks in safety that are hard to avoid.In order to develop novel vaccines with both excellent potential in eliciting antibody memory and low safety risk,it is critical to further investigate the mechanism of antibody memory in the perspective of immunology.Antibody memory is mediated by certain long-lived B cells:long-lived plasma cell can secret antibody to maintain serum antibody titer while memory B cell contributes to the rapid immune response during the secondary encounter of pathogens.Cellular and molecular processes that drive the production of long-lived plasma cells and memory B cells are subjects of intensive research and have important implications for global health.Several factors in the vaccine would indeed affect and regulate these processes,including the antigen valency,vaccine kinetics and the signal integration of both antigen and danger molecules.Many studies have focused on strategies to manipulate these factors to improve or develop new vaccines.Here,we will summarize our current knowledge on how the component in vaccines will affect their potential in generating and sustaining antibody memory,and also point out the challenges we face in the route of developing a"perfect"vaccine.

Objective To investigate whether the semen quality of men undergoing conventional semen analysis from this center has changed over time in 10 years.Methods The data of 231 519 semen samples provided by males undergoing conventional seminal anal-ysis in outpatient department of Reproductive Medicine Affiliated to Shandong University from January 2013 to December 2022 were ret-rospectively analyzed.The sperm concentration,motility,percentage of progressively motile sperm and percentage of normal morphology sperm were analyzed according to the World Health Organization(WHO)Laboratory Manual for the Examination and Treatment of Hu-man Semen(5th edition)and the changes of these parameters in 10 years were compared.Results From 2013 to 2022,there were no significant changes in the proportions of men with different sperm concentration,motility,percentage of progressively motile sperm in reproductive clinics.Approximately 27％of infertile male patients had low sperm concentration each year,approximately 34％of infer-tile male patients had low sperm motility each year,approximately 10％of infertile male patients was not found sperm in semen samples each year,and the percentages of normal morphology sperm of approximately 57％of infertile male patients were lower than the refer-ence value.The semen routine analysis results of approximately 50％of infertile male patients were in the normal range.The proportion of normal semen population did not change significantly.Conclusion The semen quality of infertile men does not change significantly in recent 10 years.

Objective:To evaluate the long-term safety, tolerability and efficacy of Lacosamide add-on therapy in Chinese children with partial-onset seizures.Methods:SP848 was a global multicenter single-arm study involving 60 Chinese children with partial-onset seizures with the age of 4-17 years who were managed by Lacosamide add-on therapy at seven hospitals across China from April 2018 to May 2019.After treatment with at least two kinds of anti-seizure medications simultaneously or sequentially, partial seizures were still poorly controlled and Lacosamide oral solution (syrup) or tablets were added.The minimum initial oral dose was 2 mg/(kg·d), and the maximum allowable dose was 12 mg/(kg·d)or 600 mg/d during the study period.The dose was adjusted according to the tolerance and seizure control level of partial-onset seizures children.Seizure frequency and the median percentage change in partial-onset seizures per 28 days from baseline to the final visit were recorded, including 50% responder rate and 75% responder rate.Results:A total of 60 Chinese children with the mean age of 9.18 (4.00-15.40) years were included in this interim analysis, involving 39 males and 21 females.The mean course of epilepsy was 5.04 (0.50-15.20) years.A total of 43 patients (71.7%) still have been treated.One patient (1.7%) has completed the 6-12 months of follow-up, and 14 patients (23.3%) have completed the follow-up for less than 6 months.The median change in the frequency of partial seizures every 28 days from baseline to the last visit was -2.91, with its median percentage as -25.46%, and the proportions of ≥50%, while ≥75% responder rate were 40.0% and 28.3%, respectively.A total of 52 patients (86.7%) had 265 treatment emergent adverse events (TEAE), 11 patients (18.3%) had 19 serious TEAE, 37 patients (61.7%) had 127 drug-related TEAE, and 11 patients (18.3%) had 16 TEAE leading to the discontinuation of the trial.The most common TEAE were upper respiratory tract infections (20 cases, 33.3%), followed by drowsiness (16 cases, 26.7%), dizziness (15 cases, 25.0%) and vomiting (13 cases, 21.7%). There were no abnormal changes in the electrocardiographic findings during the treatment.Conclusions:For Chinese patients with partial seizures who are older than the age of 4 years and poorly controlled by other drugs, Lacosamide is effective and well tolerated as an add-on therapy drug.The safety characteristics are consistent with those reported in children and adults.No new safety concerns are identified.

Objective To investigate the influence of Nei endonuclease Ⅷ-like protein 1(NEIL1)on H2O2-induced apoptosis and autophagy in lens epithelial cells(LECs).Methods Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression of NEIL1 messenger ribonucleic acid(mRNA)in the anterior lens capsule and LEC line SRA01/04 cells from age-related cataract(ARC)patients and macular epiretinal membrane patients who underwent the clear lens extraction.SRA01/04 cells were then divided into the control group,OE-Vector group and OE-NEIL1 group.RT-PCR and Western blot(WB)were used to detect the overexpression efficiency.Subsequently,the cells were divided into the control group,H2O2 group,OE-Vector H2O2 group and OE-NEIL1 H2O2 group;cell viability was detected by Cell Count-ing Kit-8,WB was used to evaluate the expression of autophagy-related proteins(ATG7,P62,Beclinl,LC3-Ⅰ and LC3-Ⅱ)and apoptosis-related proteins(Bax and Bcl-2),and fluorescent staining was adopted to measure cell apoptosis and mito-chondrial membrane potential.Results The expression of NEIL1 mRNA in the anterior lens capsule samples from ARC patients was significantly lower than that of macular epiretinal membrane patients,and the expression of NEIL1 mRNA in the SRA01/04 cells in the H2O2 group was also lower than that in the control group(both P＜0.05).The results of RT-PCR and WB revealed that the expressions of NEIL1 mRNA and protein in the SRA01/04 cells of the OE-NEIL1 group were signif-icantly higher than those in the OE-Vector group(both P=0.000).Compared with the control group,the viability of SRA01/04 cells in the H2O2 group decreased,the expression of the Bax protein was up-regulated,the Bcl-2 protein was down-regulated,viable cells labeled with Mito-Tracker decreased,and apoptotic cells labeled with Annexin V-FITC in-creased,with statistical significances(all P＜0.05).Compared with the OE-Vector H2O2 group,in the OE-NEIL1 H2O2 group,the viability of SRA01/04 cells significantly improved,the autophagy-related proteins(ATG7,P62,Beclin1,LC3-Ⅰand LC3-Ⅱ)and Bcl-2 protein were significantly up-regulated,the Bax protein was down-regulated,viable cells marked with Mito-Tracker increased,and apoptotic cells labeled with Annexin V-FITC decreased(all P＜0.05).Conclusion Under the oxidative stress induced by H2O2,NEIL1 can promote autophagy of LECs,maintain homeostasis of LECs,and then increase cell viability and reduce apoptosis of LECs,participating in the occurrence and development of ARC.