Objective To screen for potential targets of the anti-tuberculosis lead compound M6 using antibody microarray,express Mycobacterium smegmatis(Ms) target protein 2-C-methy1-D-erythriol 4-phosphate cytidylyl-transferase(IspD) in E.coli, and prepare its polyclonal antibodies, in order to provide experimental basis for in-depth study of the protein function and the development of new anti-tuberculosis drugs.Methods The minimum inhibitory concentration(MIC) of M6 against Ms and Corynebacterium glutamicum(Cg) was determined. The protein samples of Cg treated with M6 were captured and analyzed by antibody microarray, and the differential targets were identified and screened by mass spectrometry. The ispD gene of Ms was amplified by PCR and cloned into vector pET28a(+) to construct recombinant plasmid pET28a-ispD. The recombinant plasmid was transformed into E.coli BL21(DE3), and the optimal conditions for protein expression were determined by optimizing the induction temperature(16, 25, 33, 37 ℃) and IPTG concentration(0-1. 5 mmol/L). The recombinant IspD protein was purified by nickel column affinity chromatography. The purified recombinant IspD protein was used as immunogen combined with Freund's incomplete adjuvant to immunize 11 female BALB/c mice to prepare polyclonal antibodies. The titer of antiserum was determined by indirect ELISA, and the antibody specificity was detected by Western blot.Using the prepared polyclonal antibodies, the effect of M6 treatment on the expression level of Ms IspD protein was analyzed by Western blot.Results The MIC of M6 against Cg and Ms was 0. 5 and 32 μg/mL, respectively. IspD protein was one of the primary target proteins of M6. The recombinant expression plasmid pET28a-ispD was constructed correctly as identified by double enzyme digestion, and the IspD protein was expressed in E.coli. The optimal induction conditions were determined to be 1. 0 mmol/L IPTG, 16 ℃ for 14 h. After purification, the purity of recombinant IspD protein reached, and it could specifically bind to mouse anti-His tag monoclonal antibodies. The polyclonal antibodies against IspD protein achieved a high titer of 1∶102 400, which could specifically recognize IspD protein in recombinant bacterial lysate. The expression level of Ms IspD protein did not change significantly after M6 treatment.Conclusion In this study, the potential target IspD protein of M6 was successfully screened by antibody microarray screening, and Ms IspD protein was expressed in E.coli. The polyclonal antibodies against IspD were prepared by immunizing mice, providing essential experimental basic for further functional studies of this protein and the development of novel anti-tuberculosis drugs.

ObjectiveTo analyze the research status, hotspots and development trends in the application of virtual reality (VR) technology in managing negative emotions and postoperative rehabilitation of perioperative patients over the past decade. MethodsLiteratures related to the application of VR technology in managing negative emotions and postoperative rehabilitation of perioperative patients were retrieved from Web of Science Core Collection database and CNKI, covering the period from January, 2015 to August, 2025, and CiteSpace 6.3.R1 was used for bibliometric analysis. ResultsA total of 267 English literatures and 130 Chinese literatures were included, with the annual number of publications showing an upward trend. The United States was the country with the largest number of publications in English literatures, and Erasmus University Rotterdam was the institution with the largest number of publications. High-frequency keywords included virtual reality, pain, surgery, anxiety and distraction. Research hotspots mainly focused on functional exercise, negative emotions, pain management and multimodal intervention strategies. English researches were deepening towards virtual reality exposure therapy, mechanism exploration and personalized schemes, while Chinese researches focused more on the verification of rehabilitation effects. ConclusionResearches on the application of VR technology in the management of perioperative patients are rapidly developing, with research hotspots shifting from single technology application to multimodal and personalized integrated intervention. Future research should focus on exploring its intervention mechanisms, personalized schemes and the breadth of cross-departmental applications.

[摘 要] 目的：探讨复发/转移性鼻咽癌（NPC）接受含PD-1单抗免疫治疗的临床特征和预后影响因素。方法：回顾性分析2019年3月至2024年7月期间南部战区总医院确诊的95例NPC患者的临床资料和外周血生化及免疫学指标。预后分析采用Kaplan-Meier曲线，组间比较使用Log-rank检验，采用Cox比例风险模型进行单因素和多因素分析。结果：95例患者中男性81例，女性14例，中位年龄49.72岁（16~74岁），Ⅳ期91例（95.79%），所有患者均采用免疫治疗，联合或不联合化疗方案治疗，中位无进展生存期（mPFS）为10.5个月，客观缓解率（ORR）70.53%，疾病控制率（DCR）89.47%，接受含铂治疗方案患者PFS相对更长，且差异有统计学意义。紫杉醇 + 顺铂 + 氟尿嘧啶（TPF）对比吉西他滨 + 顺铂（GP）和紫杉醇 + 顺铂（TP）显示出更长的PFS，但差异无统计学意义。不同PD-1单抗治疗组间的PFS未显示出有统计学意义的差异。单因素及多因素Cox回归分析结果显示，肿瘤复发状态、初始血浆EBV感染状态、治疗周期数、基线外周血SII是复发/转移性NPC患者接受PD-1抑制剂治疗疗效预测的独立相关因素（均P < 0.05），并且非复发患者、初始血浆EBV DNA阳性、接受 ≥ 4治疗周期、基线外周血SII < 772.81的患者接受PD-1抑制剂治疗预后相对更好。结论：在接受PD-1抑制剂治疗的复发/转移性NPC患者中，非复发患者、初始血浆EBV DNA阳性、≥ 4治疗周期且外周血SII < 772.81者PFS相对更长，可早期识别免疫治疗效果不佳患者并精准干预。

High-throughput sequencing-based high-throughput screening （HTS²） technology， as a new advancement in the field of high-throughput biotechnology， is the world's first technology to integrate high-throughput sequencing into large-scale drug screening and target discovery. The artificially designed DNA probes were bound to the undetermined mRNAs of thousands of genes in cell lysates， and then the probes were ligated with ligases. The large-scale simultaneous detection of gene expression changes in thousands of drug-treated cell samples was performed using barcoding， automated operating platforms， and high-throughput sequencers. This technology enables high-throughput identification of drugs that significantly perturb the gene expression profiles characteristic of diseases. It can also take gene expression signature as the readout and exert great high-throughput advantages in the screening of multi-drug， multi-component， and multi-target drugs， as well as the research on complex mechanisms. Therefore， it is particularly suitable for elucidating the multi-target mechanisms of traditional Chinese medicine and identifying its multi-effective components. Its main technical advantages include high throughput， automation， and low cost. In recent years， HTS² technology has yielded important achievements in the elucidation of the mechanism of action of traditional Chinese medicine formulas， the scientific connotation analysis of the regional characteristics of traditional Chinese medicine， the targeted isolation of active compounds of traditional Chinese medicine， and the discovery of novel pharmacological functions of monomeric compounds of traditional Chinese medicine. In the era of artificial intelligence， HTS² technology will serve as a powerful tool for generating high-quality， original big data of traditional Chinese medicine， providing core data support and promoting AI-driven traditional Chinese medicine research. Ultimately， HTS² technology offers new strategies and critical data support for deeply analyzing the scientific connotation of traditional Chinese medicine and discovering novel traditional Chinese medicine-based drugs， thereby accelerating the modernization and internationalization of traditional Chinese medicine in China.

OBJECTIVE: To explore the clinical and genetic characteristics of two children diagnosed with two rare genetic diseases simultaneously. METHODS: Two children with comorbidity of two genetic diseases due to dual genetic mutations diagnosed at the Third Affiliated Hospital of Zhengzhou University respectively in May 2022 and March 2023 were selected as the study subjects. Clinical and genetic data of the two children were retrospectively analyzed. This study has been approved by the Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (Ethic No. 2021-062-01). RESULTS: Child 1 was a 2-year-and-4-month-old boy whose clinical manifestations included facial dysmorphism, developmental delay, short stature, microcephaly, cleft palate, cryptorchidism, hypospadias, recurrent infections and immunological abnormalities. Whole exome sequencing revealed that he had harbored a heterozygous c.6595delT (p.Y2199Ifs*65) variant of the KMT2D gene and a heterozygous c.1892G>A (p.R631Q) variant of the PIK3R1 gene. This has led to a dual genetic diagnosis of Kabuki syndrome and PI3Kδ-related immunodeficiency type 36. Child 2 was a 15-year-old girl whose clinical manifestations included epilepsy, Albright's hereditary osteodystrophy, long body trunk, short limbs, hypocalcemia, hyperphosphatemia and hyperparathyroidism. The child also had a family history of short stature. Whole exome sequencing revealed that she had harbored a heterozygous c.2T>C (p.Met1?) variant of the GNAS gene and deletion of exons 2 to 6 of the SHOX gene. The two variants have led to dual diagnose of pseudohypoparathyroidism and X-linked idiopathic short stature. CONCLUSION When the clinical phenotype of a genetic disease is complex and cannot be fully explained with a single genetic variant, multiple pathogenic variants should be considered, and this may lead to the diagnosis of co-morbid genetic diseases. To adopt or supplement corresponding genetic testing in time and re-analyze the genetic data may facilitate accurate diagnosis of co-morbid genetic diseases.
Child, Preschool ; Female ; Humans ; Male ; Class Ia Phosphatidylinositol 3-Kinase/genetics* ; Comorbidity ; Exome Sequencing ; Mutation ; Rare Diseases/genetics* ; Retrospective Studies ; Adolescent

Objective To analyze the effect of Shufeng Tongqiao decoction combined with methylprednisolone tablets in patients with chronic sinusitis and nasal polyp(CRSwNP)after operation.Methods A total of 104 patients with CRSwNP underwent endoscopic nasal surgery were prospectively selected,and patients were randomly divided into the hormone group(n=52,postoperative methylprednone tablets)and the study group(n=52,postoperative combined treatment with Shufeng Tongqin Decoction).After removal and shedding,50 cases were included in the hormone group,and 49 cases were included in the study group.The curative effect,lung wind-heat syndrome score,nasal mucosa recovery(assessed by Lund-Kennedy score system)and nasal mucosa remodeling were compared between the two groups.Serum levels of human β-defensin(HBD)-2 and high mobility group protein B1(HMGB1)were detected by enzyme-linked immunosorbent assay.Results The total control rate was higher in the study group than that of the hormone group(95.92%vs.80.00%,P＜0.05).At 12 weeks after surgery,the lung meridian wind heat syndrome scores,Lund-Kennedy scores and serum HMGB1 level were lower than before surgery in both groups(P＜0.05),and the study group was lower than the hormone group(P＜0.05).At 12 weeks after surgery,the epithelial injury grading was better in the study group than that in the hormone group(P＜0.05),and the basement membrane thickness was thinner than that in the hormone group(P＜0.05).Conclusion The joint of Shufeng Tongqiao decoction and methylprednisolone tablets has a better effect on postoperative CRSwNP patients,and it can greatly improve the levels of serum hBD-2 and HMGB1,effectively inhibit nasal mucosal remodeling,and greatly improve the recovery of nasal mucosa.

Objective:To investigate the correlation between non-Hodgkin lymphoma (NHL) and chronic hepatitis B virus (HBV) infection by using the method of two-sample bidirectional Mendelian randomization (MR) analysis.Methods:Genetic variation data for NHL came from the Finnish database (FinnGen) Consortium 2021 public genome-wide association study (GWAS) dataset including 1 088 patients with NHL and 299 952 control subjects. The GWAS dataset for chronic HBV infection was derived from GWAS analysis published in 2021, including 145 NHL patients and 351 740 control subjects. NHL was used as an exposure factor, single nucleotide polymorphism (SNP) significantly associated with NHL was used as an instrumental variable (IV), chronic HBV infection was used as an outcome variable. The two-sample MR analysis was performed by using inverse-variance weighted (IVW) method. Chronic HBV infection was taken as an exposure factor, SNP significantly associated with chronic HBV infection was taken as IV, and NHL was taken as outcome variable, and then reverse two-sample MR analysis was performed. The IVW method used the inverse variance of each IV as the weight to fit, and the ratio method was used to measure SNP one by one and make weighted regression analysis, so as to obtain the overall estimate. MR-Egger regression and the weighted median (WME) method were also used to supplement the IVW method. In sensitivity analysis, leave-one-out sensitivity analysis was used to evaluate the impact of a single SNP. Cochran Q test was used to analyze the heterogeneity of the selected IV. MR-Egger regression was used to measure the average horizontal pleiotropy of IV, and the P-value of directivity was calculated. The MR-pleiotropy residual sum and outlier (MR-PRESSO) Global Test was used to exclude possible horizontal pleiotropic outliers and reduce bias. Results:In the leave-one-out sensitivity analysis, SNP with significant effects on causal associations was excluded. In forward MR analysis, IVs were 10 SNPs associated with NHL; the IVW method indicated that there was no causal association between NHL and chronic HBV infection ( OR = 0.979, 95% CI: 0.925-1.036, P = 0.465). MR-Egger regression ( OR = 0.992, 95% CI: 0.926-1.062, P = 0.825) and WME method ( OR = 0.992, 95% CI: 0.934-1.055, P = 0.805) were used as supplementary methods to obtain the consistent results. In sensitivity analysis, Cochran Q test showed no heterogeneity among IVs (IVW method: P = 0.271, MR-Egger regression: P = 0.239). Horizontal pleiotropy was not found in the MR-Egger regression (intercept was -0.01, P = 0.778) and the MR-PRESSO Global Test ( P > 0.05), suggesting robust results. In the reverse MR analysis, IVs were 8 SNPs associated with NHL; the IVW method ( OR = 1.117, 95% CI: 0.942-1.324, P = 0.202) also found no significant causal relationship between chronic HBV infection and NHL; MR-Egger regression ( OR = 0.777, 95% CI: 0.450-1.343, P = 0.401) and WME method ( OR = 1.120, 95% CI: 0.887-1.415, P = 0.351) also showed similar risk estimates. Sensitivity analysis also suggested the consistency and reliability of the results. Cochran Q test showed no heterogeneity among IVs (IVW method: P = 0.775, MR-Egger regression: P = 0.903). Horizontal pleiotropy was not found by MR-Egger regression (intercept was 0.102, P = 0.548) and MR-PRESSO Global Test ( P > 0.05). Conclusions:MR analysis suggests no causal relationship between NHL and chronic HBV infection.

Objective To explore the levels of Kruppel like factor 5(KLF5)and Dickkopf related protein 1(DKK1)in the serum of patients with primary liver cancer and analyze their diagnostic value.Methods A to-tal of 98 patients with primary liver cancer diagnosed in the hospital from March 2021 to March 2024 were in-cluded as the experimental group,and they were divided into early group(40 cases)and late group(58 cases)according to the severity of the disease.Meantime,101 patients with benign liver disease admitted to the hos-pital during the same period time were selected as the control group,and 100 healthy individuals who under-went the physical examination during the same period were selected as the healthy group.Serum samples were obtained from the groups and general clinical data were collected and organized.Enzyme linked immunosor-bent assay was applied to detect the levels of KLF5 and DKK1,and the differences and changes in serum KLF5 and DKK1 levels among the groups were analyzed.Multivariate Logistic regression was applied to ana-lyze the influencing factors of occurrence of primary liver cancer.Receiver operating characteristic curve was used to analyze the efficacy of serum KLF5 and DKK1 levels in the diagnosis of primary liver cancer.Results The levels of alpha fetoprotein(AFP),carcinoembryonic antigen(CEA),KLF5 and DKK1 in the ex-perimental group were significantly higher than those in control group and healthy group(P＜0.05).The lev-els of serum KLF5 and DKK1 were obviously different in patients with primary liver cancer with or without lymph node metastasis(P＜0.05).The levels of serum KLF5 and DKK1 in late group were higher than those in early group(P＜0.05).AFP,CEA,KLF5 and DKK1 levels were independent risk factors for primary liver cancer(P＜0.05).The area under the curve of the combined diagnosis of serum KLF5 and serum DKK1 was 0.928,with specificity of 93.10％,which was better than that of the individual diagnosis(Z combination-DKK1=2.186,Z combination-KLF5=3.386,P=0.029,P=0.001).Conclusion The levels of KLF5 and DKK1 in the serum of patients with primary liver cancer are obviously elevated and change with changes in the condition of dis-ease.The combined detection of KLF5 and DKK1 is of great significance for the diagnosis of primary liver cancer.

Objective:To evaluate the clinical efficacy of the modified medial gastrocnemius myocutaneous flap (MGMF) with extended anterior, posterior and (or) inferior boundaries.Methods:From January 2002 to September 2022, modified MGMFs were applied onto 33 patients who received reconstructive surgery for soft-tissue defects around knee or in calf, in the Department of Orthopaedics, the Second Xiangya Hospital of Central South University. The size of defects ranged from 10 cm×4 cm to 22 cm×12 cm, and the flap size ranged from 15 cm×6 cm to 28 cm×14 cm. Twenty-five patients had the complication of chronic osteomyelitis. The boundaries of a modified MGMF were as follows: the anterior boundary was the anterior border of the tibia, where the posterior boundary at 3.0 cm lateral to the posterior midline, the proximal boundary at the popliteal fossa crease, and the distal boundary at the plane 2.0 cm above the tip of medial malleolus. The anterior edge of the modified MGMF was designed running along the medial edge of the defect and its curved extension line. Pretibial skin was equally divided into 9 zones, with the 1st to 9th zones from proximal to distal in sequence. Postoperative routine anti-infection treatment was offered. All patients were included in the postoperative follow-up through outpatient visits, telephone or WeChat interviews. Flap viability and wound healing in both donor and recipient sites were evaluated. Function of the affected limb was assessed using the evaluation criteria established by Punor et al.Results:All patients were included in the follow-up for 1 to 169 (median duration: 9)months. The 33 modified MGMFs included MGMFs with extended boundary of anterior ( n=18), inferior ( n=5), anterior combined with inferior ( n=6), posterior combined with anterior ( n=2), and posterior combined with inferior ( n=2) boundaries. Twenty-nine (87.9%) flaps survived completely. Partial necrosis occurred in 4 flaps(12.1%)(2 flaps with extended anterior boundary and 2 flaps with extended inferior boundary). The anterior margins of 26 flaps (78.7%) with extended anterior boundary alone or in combination with extended inferior or posterior boundary exceeded the medial edge of the tibia by 1.0-4.5 (mean, 2.1) cm, and 3 of them reached the anterior edge of tibia. Fourteen (42.4%) modified MGMFs were used to reconstruct the defects involving 1/3 of distal calf, and the distal ends of these defects were located in the 7th ( n=8) or 8th ( n= 6) zone. All the skin grafts in the donor sites survived. During follow-up, 31 patients (93.9%) showed no sign of infection, and 2 patients (6.1%) who had recurrence of chronic osteomyelitis. Functions of the affected limbs were excellent ( n=25), good ( n=6) and fair ( n=2) by Punor et al. Conclusion:Modified MGMF with extended anterior, posterior and (or) inferior boundaries is clinically feasible. It offers advantages of easier design and operation. It can be used to reconstruct a more distal, wider and larger defect as well as broadens the application of the MGMF.

Objective:This current study aims to explore the approaches for constructing a professional clinical research system within the context of research ward development, with the ultimate objective of providing valuable guidance for the establishment and development of proficient clinical research teams.Methods:Through a comprehensive case analysis, integrating the practical experiences from clinical trials conducted in the research ward of a Class-A tertiary hospital in Beijing, along with an extensive review of relevant literature and policy studies, this paper examined the current state of domestic clinical research implementation teams. Subsequently, a series of strategies were devised to build and foster professional clinical research teams and to explore corrective measures for cultivating a dynamic professional clinical research talent ecosystem.Results:The development of full-time clinical research teams in China was rather slow, and there was a lack of mature clinical trial teams training blueprints. Drawing on the practical experience accumulated during the establishment of a professional clinical research team in a leading hospital in Beijing, it was crucial to attach utmost importance to the optimal allocation of human and material resources. This required the systematic training of principal investigators, coordinating researchers, and research assistants, as well as the setting up of a comprehensive support system, an advanced scientific research team, and a quality control unit. Moreover, the standardization of operational models of both domestic and foreign research institutions, along with the implementation of corresponding support and incentive mechanisms, and the strengthening of training and continuing education frameworks were equally significant.Conclusions:During the process of assembling a full-time clinical research team, it is of utmost significance to cultivate professional principal investigators, coordinating researchers, and research assistants. Complemented by the establishment of a comprehensive support team, a scientific research team, and a quality control team, along with corresponding support and incentive mechanisms, this is crucial for constructing a professional clinical research execution team and a sustainable talent ecosystem in the research ward. Eventually, this will drive the efficient and high-quality progress of China's pharmaceutical industry.