ObjectiveTo perform a genome-wide association study of rubella virus vaccine strain BRD-Ⅱ, so as to fully grasp the sequence characteristics of this genome. MethodsSecond-generation sequencing method was used to conduct the whole-genome sequencing on the vaccine strain BRD-Ⅱ, and the affinity tree of this genome with some vaccine strains and wild-type rubella virus strains was analyzed using the maximum likelihood method. The average genetic distance of nucleic acid sequence of each vaccine strain protein was determined. And homology comparison of structural proteins of each rubella vaccine strain, plus the comparison between this genome with the AY258323.1 genome sequence, were conducted to analyze the homology of E1 protein between the wild-type rubella virus reference strain and vaccine strain BRD-Ⅱ. ResultsThe sequencing results showed that the BRD-Ⅱ strain was a single-molecule single-stranded positive-strand ribonucleic acid (RNA), composed of 9 778 nucleotides, with a GC content of 69.35 %. The C protein was composed of 300 amino acids, the E2 glycoprotein was composed of 282 amino acids, and the E1 glycoprotein was composed of 481 amino acids. The results of preliminary analysis showed that the average genetic distances of nucleic acid sequences were 0.066 700 for the P150 protein, 0.061 933 for the P90 protein, 0.057 850 for the C protein, 0.068 167 for the E2 protein, and 0.068 833 for the E1 protein, respectively. The amino acid sequences in the E2 protein and E1 protein regions of the two BRD-Ⅱ strains did not change, confirming the conserved regions of the E1 protein by comparison. ConclusionThe sequence characteristics of the genome are clarified, which have laid a broad foundation for the subsequent detection of the genetic stability of the main antigen genes.

ObjectiveThe effects of two microbial fertilizers （Bacillus subtilis fertilizer and Trichoderma harzianum-Purpureocillium lilacinum compound fertilizer） on the growth and development， the accumulation of active ingredients， and the microbial community diversity of rhizosphere soil of Atractylodes chinensis were investigated. MethodsA field experiment was carried out with two-year-old Atractylodes chinensis as the test material. Plant samples were collected during the wilt stage （September 26， 2023） to determine the general agronomic traits of Atractylodes chinensis. High-performance liquid chromatography （HPLC） was utilized to evaluate the effects of microbial fertilizers on the synthesis and accumulation of four active ingredients （atractylodin， atractylon， β-eudesmol， and atractylenolide Ⅰ） in Atractylodes chinensi. PacBio Sequel sequencing technology was used to explore the differences in bacterial community structures and diversity in the rhizosphere soil of Atractylodes chinensis treated with different microbial fertilizers. ResultsThe two microbial fertilizers had significant growth-promoting effects on Atractylodes chinensis. Compared with those of the CK group， the stem diameter， stem and leaf dry and fresh weight， and rhizome dry and fresh weight of Atractylodes chinensis significantly increased by 0.47-1.07 times （P<0.05） after the application of the Bacillus subtilis fertilizer （16 kg/667 m²）， and those significantly increased by 0.62-0.96 times （P<0.05） after the application of the Trichoderma harzianum-Purpureocillium lilacinum compound fertilizer （1.5 kg/667 m²）. The effect on plant height was not significant. The application of two microbial fertilizers was beneficial to the accumulation of atractylodin， atractylon， β-eudesmol， and atractylenolide Ⅰ （P<0.01）， and the effect of the Bacillus subtilis fertilizer on the accumulation of active ingredients of Atractylodes chinensis was better than that of the Trichoderma harzianum-Purpureocillium lilacinum compound fertilizer. The results of high-throughput sequencing showed that compared with the CK group， the Bacillus subtilis fertilizer （8 kg/667 m²） could significantly increase the diversity of rhizosphere bacterial species by regulating the Simpson index and Shannon index （P<0.05）， and the Trichoderma harzianum-Purpureocillium lilacinum compound fertilizer significantly reduced the bacterial diversity （P<0.05）. The relative abundance of dominant bacteria was compared at the phylum and genus levels. The relative abundance of Proteobacteria （45.73%） and Burkholderia_Caballeronia_Paraburkholderia （9.98%） significantly increased after the application of the Bacillus subtilis fertilizer （P<0.01）， and the relative abundance of Acidobacteriota （20.53%） and Sphingomonas （3.63%） increased significantly （P<0.01） after the application of the Trichoderma harzianum-Purpureocillium lilacinum compound fertilizer. The relative abundance of beneficial bacteria in the Bacillus subtilis fertilizer was slightly higher than that in the Trichoderma harzianum-Purpureocillium lilacinum compound fertilizer. Pearson correlation analysis showed that Burkholderia_Caballeronia_Paraburkholderia and Sphingomonas were positively correlated with the content of atractylodin， atractylon， β-eudesmol， and atractylenolide Ⅰ （P<0.05）. ConclusionThe application of the Bacillus subtilis fertilizer and Trichoderma harzianum-Purpureocillium lilacinum compound fertilizer can increase the yield of medicinal materials and promote the synthesis and accumulation of active ingredients by regulating the rhizosphere microecological diversity of Atractylodes chinensis， and the application effect of the Bacillus subtilis fertilizer is better than that of the Trichoderma harzianum-Purpureocillium lilacinum compound fertilizer.

The Janus kinase/signal transducers and activators of transcription (JAK-STAT) control natural killer (NK) cells development and cytotoxic functions, however, whether long non-coding RNAs (lncRNAs) are involved in this pathway remains unknown. We found that miR155HG was elevated in activated NK cells and promoted their proliferation and effector functions in both NK92 and induced-pluripotent stem cells (iPSCs)-derived NK (iPSC-NK) cells, without reliance on its derived miR-155 and micropeptide P155. Mechanistically, miR155HG bound to miR-6756 and relieved its repression of JAK3 expression, thereby promoting the JAK-STAT pathway and enhancing NK cell proliferation and function. Further investigations disclosed that upon cytokine stimulation, STAT3 directly interacts with miR155HG promoter and induces miR155HG transcription. Collectively, we identify a miR155HG-mediated positive feedback loop of the JAK-STAT signaling. Our study will also provide a power target regarding miR155HG for improving NK cell generation and effector function in the field of NK cell adoptive transfer therapy against cancer, especially iPSC-derived NK cells.

Objective To investigate the efficacy and safety of camrelizumab monoclonal antibody combined with molecular-targeted therapy in elderly patients with unresectable or advanced hepatocellular carcinoma(HCC).Methods A retrospective analysis was performed for the patients with unresectable/advanced HCC who attended six hospitals from January 1,2019 to March 31,2021,and all patients received camrelizumab monoclonal antibody treatment,among whom 84.8%also received targeted therapy.According to the age of the patients,they were divided into elderly group(≥65 years)and non-elderly group(＜65 years).The two groups were assessed in terms of overall survival(OS),progression-free survival(PFS),objective response rate(ORR),disease control rate(DCR),and immune-related adverse events(irAE).The chi-square test or the Fisher's exact test was used for comparison of categorical data between groups;the independent samples t-test was used for comparison of normally distributed continuous data,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups.The Kaplan-Meier method was used for survival analysis,and the log-rank test was used for comparison of survival curves.Univariate and multivariate Cox proportional hazards regression analyses were used to determine the independent influencing factors for PFS and DCR at 6 months.Results A total of 99 HCC patients were enrolled,with 27 in the elderly group and 72 in the non-elderly group.The elderly group had an OS rate of 67.8%,an ORR of 44.4%,and a DCR of 74.1%at 12 months and a median PFS of 6.4(95%confidence interval[CI]:3.0-12.4)months,with no significant differences compared with the non-elderly group(all P＞0.05).The median OS was unavailable for the elderly group,while the non-elderly group had an OS of 18.9(95%CI:13.0-24.8)months;there was no significant difference between the two groups(P=0.485).The univariate and multivariate Cox regression analyses showed that major vascular invasion(MVI)was an independent risk factor for PFS(hazard ratio[HR]=2.603,95%CI:1.136-5.964,P=0.024)and DCR(HR=3.963,95%CI:1.671-9.397,P=0.002)at 6 months,while age,sex,etiology of HBV infection,presence of extrahepatic metastasis,Child-Pugh class B,and alpha-fetoprotein＞400 ng/mL were not associated with PFS or DCR at 6 months.For the elderly group,the incidence rates of any irAE and grade 3/4 irAE were 51.9%and 25.9%,respectively,with no significant differences compared with the non-elderly group(P＞0.05),and skin disease was the most common irAE in both groups(39.4%).Conclusion Camrelizumab monoclonal antibody combined with molecular-targeted therapy has similar efficacy and safety in patients with unresectable/advanced HCC aged≥65 years and those aged＜65 years.MVI is associated with suboptimal response to immunotherapy and poor prognosis.

OBJECTIVE To explore the effect of Huangqin decoction on improving peritubular fat inflammatory microenvironment and protecting vascular endothelial function in obese hypertensive rats by regulating the NEK7-NLRP3/IL-1β inflammatory axis.METHODS Fifty 4-week-old male Wistar rats were selected,10 of which were randomly selected as the control group,and the oth-er 40 were fed a high-salt and high-fat diet to establish an obese hypertension model.The rats with successful modeling(20 rats)were randomly divided into the model group,normal-dose Huangqin decoction group,high-dose Huangqin decoction group,and IL-1β in-hibitor group,with 5 rats in each group.From the 12th week,the normal-dose group was gavaged with Huangqin decoction 2.835 g·kg-1,the high-dose group was gavaged with Huangqin decoction 5.67 g·kg-1,and the IL-1β inhibitor group was intraper-itoneally injected with 1.5 mg·kg-1 AS101,3 times a week,for 8 weeks.The rats were weighed and blood was collected 12 h after the last administration,and the thoracic aorta and perivascular fat tissue were isolated.Serum inflammatory factors were detected,patho-logical changes were observed,eNOS expression was detected by immunofluorescence,and NEK7,NLRP3,Caspase-1,ASC,and IL-1β expression levels were detected by Western blot and qPCR.RESULTS The rats in the model group had a significant increase in body weight,an increase in the area of peritubular fat lipid droplets,and severe endothelial injury;systolic blood pressure,diastolic blood pressure,serum IL-1β,IL-6,and TNF-α were significantly elevated in the model group,and the expression of eNOS was sig-nificantly reduced,and the expression levels of NEK7,NLRP3,Caspase-1,ASC,and IL-1β proteins and mRNAs were significantly elevated.Compared with the model group,rats in the Huangqin decoction and IL-1β inhibitor groups had lower body weights,reduced endothelial damage,lower systolic and diastolic blood pressures,lower serum IL-1β,IL-6,and TNF-α,and higher eNOS expression.NEK7,NLRP3,Caspase-1,ASC and IL-1β protein expression was significantly reduced in the high dose group of Huan-gqin decoction and the IL-1β inhibitor group.In addition,Huangqin decoction protected the endothelial function of obese hypertensive vessels in a dose-dependent manner,with the effect being more pronounced in the high-dose group.CONCLUSION Huangqin de-coction can improve the inflammatory microenvironment of perivascular fat and protect the vascular endothelial function in obese hyper-tension by regulating the NEK7-NLRP3/IL-1β inflammatory axis.

Objective:This study aimed to explore the status of radiological Kashin-Beck disease (KBD) among school-aged children in Chamdo City, Tibet, through a 3-year monitoring survey, providing epidemiological evidence for prevention and control strategies.Methods:The target areas for this study were Luolong, Bianba, and Basu counties in Chamdo City, Tibet Autonomous Region, identified as having the most severe historical cases of KBD. Children aged 7-12 years attending school were enrolled as study subjects. Anteroposterior X-ray films of the right-hand were taken, and radiological diagnoses were made based on the "Diagnosis of Kashin-Beck Disease" criteria (WS/T 207-2010). Two experienced researchers independently reviewed the X-rays, and intra- and inter-group consistency were assessed using weighted Kappa values and percentage agreement. Cross-sectional surveys were conducted in 2017 and 2020 to describe the X-ray detection rates of KBD, and logistic regression analysis was employed to construct a predictive model of risk factors for radiological KBD cases.Results:In 2017, a total of 5,711 children aged 7-12 years in Chamdo City, Tibet, participated in the baseline cross-sectional survey (average age 9.2 years, 48.0% female), with 28 cases of radiological KBD. The age- and gender-standardized prevalence rate was 0.527%. In 2020, 6,771 participants (average age 9.3 years, 49.5% female) underwent a second cross-sectional survey, with 9 cases of radiological KBD and a standardized prevalence rate of 0.134%. Logistic regression analysis indicated that older age [ OR=2.439, 95% CI(1.299, 4.580), P=0.006] and female gender [ OR=8.157, 95% CI(1.016, 65.528), P=0.048] were independent risk factors for radiological KBD cases. Conversely, higher residential altitude, under the premise of Tibet's high altitude, was a protective factor [ OR=0.995, 95% CI(0.990, 0.999), P=0.032). Conclusion:The radiographically positive detection rate of KBD among school-aged children in Chamdo City, Tibet Autonomous Region, is at an extremely low level and showing a declining trend, reaching the historical standard in 2020. Considering the absence of positive signs in affected children, it suggests that local KBD has been effectively eliminated.

Objective:To investigate the application value of virtual reality laparoscopic simulator training in laparoscopic transabdominal preperitoneal hernioplasty (TAPP).Methods:The prospective cohort study was conducted. Twenty young physicians from The First Affiliated Hospital of Harbin Medical University with ≥3 years of clinical experience in general surgery and no foundation in laparoscopy were selected for training during July to August 2023. Physicians were divided into two groups based on random number table method. Physicians undergoing virtual reality laparoscopic simulator training were divided into the virtual reality group, and physicians undergoing regular laparoscopic simulator training were divided into the regular group. Two groups of physicians were trained using laparoscopic simulator for 10 days (2 hours for each skill, with a total of 6 hours per day for 3 skills), and the training covered basic laparoscopic surgical skills such as clamping, cutting and suturing and knotting. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups were conducted using the independent sample t test and the paired ttest was used for intra group comparison. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers, and comparison between groups were conducted using the chi-square test. Results:(1) Physicians situation before training. A total of 20 physicians were selected for eligibility. There were 13 males and 7 females, aged 31(range, 30?34)years. There are 7 males and 3 females in the virtual reality group, with age of (31.5±1.4)years and the TAPP simulation surgery time of (42±4)minutes before training. There are 6 male and 4 female students in the regular group, with age of (31.2±1.0)years and the TAPP simulation surgery time of (42±4)minutes before training. There was no significant difference in gender, age, TAPP simulation surgery time between the two groups of physicians ( P>0.05), confounding bias ensured comparability. (2) Basic skills of physicians before and after training. For physicians in the virtual reality group, the clamping score was 5.1±1.0, the cutting score was 4.9±1.0, the suturing and knotting score was 4.7±1.5 before training. The clamping score was 8.0±1.3, the cutting score was 7.9±1.5, the suturing and knotting score was 6.6±1.3 after training. There were significant differences in the above indicators before and after training ( t=?5.75, ?5.21, ?3.07, P<0.05). For physicians in the regular group, the clamping score was 5.3±1.0, the cutting score was 5.0±1.2, the suturing and knotting score was 4.3±1.5 before training. The clamping score was 7.1±1.2, the cutting score was 6.7±1.3, the suturing and knotting score was 5.7±1.1 after training. There were significant differences in the above indicators before and after training ( t=?3.73, ?3.16, ?2.42, P<0.05). (3) Completion of simulated surgical situations before and after training. The time of completing TAPP simulation surgery for virtual reality group after training was (29±3)minutes, versus (42±4)minutes before training, showing a significant difference before and after training ( t=7.69, P<0.05). The time of completing TAPP simulation surgery for regular group after training was (36±4)minutes, versus (42±4)minutes before training, showing a significant difference before and after training ( t=3.75, P<0.05). The time of completing TAPP simulation surgery of virtual reality group after training was shorter than that of regular group, showing a significant difference between the two groups ( t=?3.89, P<0.05). Conclusion:Both of virtual reality and regular laparoscopic simulator can enhance the basic laparoscopic surgical skills and the proficiency of TAPP simulation surgery of physicians, and the training effect of virtual reality laparoscopic simulators is better.

Objective:Bone morphogenetic protein-4(BMP4)has been proved to be an important regulatory factor for the pathological process of atherosclerosis(AS).However,there are few related clinical studies.This study aims to investigate the levels of plasma BMP4 in patients suffering from the arterial occlusive diseases(ACD)characterized by AS,and further to test the relationship between BMP4 and inflammation and vascular injury. Methods:A total of 38 ACD patients(the ACD group)and 38 healthy people for the physical examination(the control group)were enrolled.The plasma in each subject from both groups was obtained to test the levels of BMP4,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-10,and vascular endothelial cadherin(VE-cadherin),and the relationship between BMP4 and the detected indicators above were further analyzed. Results:Compared with the control group,the patients in the ACD group displayed significant elevations in the neutrophil to lymphocyte ratio[NLR,1.63(1.26,1.91)vs 3.43(2.16,6.61)]and platelet to lymphocyte ratio[PLR,6.37(5.26,7.74)vs 15.79(7.97,20.53)],while decrease in the lymphocyte to monocyte ratio[LMR,5.67(4.41,7.14)vs 3.43(2.07,3.74)](all P<0.05).Besides,the ACD patients displayed significant elevations in plasma BMP4[581.26(389.85,735.64)pg/mL vs 653.97(510.95,890.43)pg/mL],TNF-α[254.16(182.96,340.70)pg/mL vs 293.29(238.90,383.44)pg/mL],and VE-cadherin[1.54(1.08,2.13)ng/mL vs 1.85(1.30,2.54)ng/mL],and decrease in IL-10[175.89(118.39,219.25)pg/mL vs 135.92(95.80,178.04)pg/mL](all P<0.05).While the levels of IL-1β remained statistically comparable between the 2 groups(P=0.09).Furthermore,the plasma BMP4 levels were further revealed to be positively correlated with the levels of IL-1β(r=0.35),TNF-α(r=0.31)and VE-cadherin(r=0.47),while they were negatively correlated with the levels of IL-10(r=-0.37;all P<0.01). Conclusion:After ACD occurrence,the patients'plasma concentrations of BMP4 would be upregulated,which may serve as a candidate to indicate the levels of inflammation and vascular injury.

Objective:To analyze the clinical manifestations, laboratory results and treatment outcomes of juvenile localized scleroderma(JLS).Methods:This was a retrospective case series study.Epidemiological and clinical data of patients with JLS treated in the Department of Dermatology, Beijing Children′s Hospital, Capital Medical University from January 2019 to August 2021 were analyzed.Results:Among the 228 children, there were 95 males and 133 females, with a male-to-female ratio of 1.0∶1.4.The median age of onset was 67 months(5.6 years), and the time from onset to diagnosis was 1 month to 106 months.Eight patients(3.5%) had suspected predisposing factors before onset, 25 patients(11.0%) had other diseases at the same time, and 3 patients(1.3%) had a family history of autoimmune diseases.The most common clinical type of JLS was circumscribed morphea(101 cases, 44.3%), followed by linear scleroderma(91 cases, 39.9%), generalized morphea(31 cases, 13.6%), mixed scleroderma(23 cases, 10.1%) and pansclerotic morphea(2 cases, 0.9%).Among them, 76 patients(46.3%) were antinuclear antibody(ANA) positive, and 14 patients(15.0%) were extractable nuclear antigen antibody(ENA) positive.Ninety-one of patients with circumscribed morphea were treated with medium and strong corticosteroids, vitamin D3 derivatives or calcineurin inhibitors.Patients of other types received systemic corticosteroid treatment, with an initial dose range of 1-2 mg/kg and a maximum dose of 60 mg/d.Among them, 72 patients were additionally treated with Methotrexate, with an initial dose range of 10-15 mg/m 2, once a week, and 9 patients were additionally treated with biological agents.The follow-up results showed that the skin symptoms of the patients who were followed up in the dermatology outpatient department had improved to a certain extent and could remain inactive. Conclusions:Children with JLS in the dermatology department are mainly preschool- and school-age.Circumscribed morphea is the most common type, mainly treated with glucocorticoids, vitamin D3 derivatives or calcineurin inhibitors.No specific laboratory test index is found.Corticosteroids combined with Methotrexate are recommended for systematic treatment of other types of JLS.

Objective:To investigate the accuracy of magnetic resonance imaging (MRI) cine sequences in determining the range of tumor motion in radiotherapy, providing a basis for the precise delineation of the target volume in motion for radiation therapy.Methods:A modified chest motion phantom was placed in a MRI scanner, and a water-filled sphere was used to simulate a tumor. True fast imaging with steady precession (TrueFISP) MRI cine sequences from Siemens were used to capture the two-dimensional motion images of the simulated tumor. The phantom experiments were divided into three modes: head-foot motion mode, rotation motion mode, and actual respiratory waveform mode. In the head-foot motion mode, respiratory motion period (3, 4, 5, 6, 7 and 8 s), amplitude (5, 10 and 15 mm), and respiratory waveform of the simulated tumor (sin and cos4) were set, resulting in a total of 36 motion combinations. In the rotation motion mode, a cos4 waveform was used for respiration, with respiratory periods of 3, 4, 5, 6, 7 and 8 s, head-foot motion set amplitudes of 5, 10 and 15 mm, and anterior-posterior (AP) and left-right (LR) motion set amplitudes in three combinations ([2.5, 2.5] mm, [2.5, 5.0] mm, [5.0, 5.0] mm), resulting in a total of 54 motion combinations. In the actual respiratory waveform mode, respiratory waveforms of 5 randomly selected patients from Affiliated Cancer Hospital of Zhengzhou University were obtained. Under each motion combination, TrueFISP cine images (30 frames, with an acquisition time of 11 s per frame) were obtained. The code was used to automatically identify the two-dimensional coordinates of the center of the simulated tumor in each image, and sin and cos4 functions were separately employed to fit the tumor position in the motion direction, thereby obtaining the fitted motion period and amplitude. The difference between the maximum and minimum values of the tumor's center coordinates in the head-to-foot direction is taken as the range of movement, referred to as the calculated amplitude. For the actual respiratory waveform, the distance between the measured maximum and minimum positions is used to calculate the amplitude.Results:In the head-foot motion mode, the fitted amplitudes of both sin and cos4 waveforms deviated from the set amplitudes by 0-0.51 mm, with relative deviations of 0%-4.2%. The deviation range between the calculated amplitudes and the set amplitudes of the two waveforms were 0.08-0.94 mm, with relative deviations of 1.1%-6.3%. In the rotation motion mode, the fitted amplitudes deviated from the set amplitudes by 0-0.61 mm, with relative deviations of 0%-6.2%. And the deviation range between the calculated amplitudes and the set amplitudes were 0.16-0.94 mm, with relative deviations of 0%-6.3%. In the actual respiratory waveform motion mode, the deviation range between the calculated amplitudes and the set amplitudes were 0.10-0.48 mm, with relative deviations of 2.2%-8.6%.Conclusion:TrueFISP cine sequences show minimal deviations in determining the range of tumor head-foot motion and effectively captures the tumor's movement state, thereby providing important support for the precise definition of the tumor movement target area during radiotherapy .