OBJECTIVE: The rat model of collagen-induced arthritis (CIA) was successfully established to investigate the therapeutic effect of wasp venom extract on rheumatoid arthritis (RA) rats. METHODS: The rats were randomly divided into normal group, model group, positive control group (bee venom lyophilized powder for injection, 1.25 mg•kg-1) and low, medium and high dose (0.125, 0.25, 0.5 mg•kg-1) of wasp venom extract groups, in addition the normal group, the other group rats were induced by chicken type Ⅱ collagen and complete Freund′s adjuvant which was multi-point injected into the left hindfoot of rats, once every 7 d for 14 d. After successful modeling, the rats were given corresponding dose of drugs continuous administration for 14 d. The diameter and perimeter of the ankle joint and AI score of the rats were measured before modeling, on the 14th day of modeling and on the 14th of administration, respectively; The changes of organ index and HE staining in ankle tissue were observed. The contents of inflammatory factor (IL-1β, IL-6, TNF-α, IL-8, PGE-2, COX-2) and rheumatoid factors (IgG, IgA, IgM) in serum of rats were detected by ELISA. The changes of T lymphocyte subsets in spleen tissue of rats were detected by flow cytometry. RESULTS: Compared with the model group, the extract of wasp venom had significant inhibitory effect on joint swelling (diameter and perimeter) of CIA rats (P<0.01), The joint index scores of CIA rats was decreased(P<0.01 or P<0.05). The organs index of rats restored to varying degrees (P<0.01 or P<0.05), to improved pathological structure of ankle joint, and decreased the expression of inflammatory and rheumatoid factors in serum of rats (P<0.01 or P<0.05), to regulate and improve the proportion of T lymphocyte subsets(P<0.01 or P<0.05). CONCLUSION: The extract of wasp venom has significant therapeutic effect on CIA rats, which is related to its regulation of inflammatory cytokine network and immunity.

OBJECTIVETo determine the allelic sequences and genetic polymorphism of six short tandem repeats (STRs) loci on Y chromosome, including DYS441, DYS442, DYS443, DYS444, DYS445 and DYS446 in Chinese Korean ethnic males from Yanbian region of Jilin province, China, and to construct a preliminary database.

METHODSAllele frequencies of the six STR loci in 205 Chinese Korean ethnic individuals were analyzed by multiplex polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE).

RESULTSRespectively, 8, 7, 7, 5, 6 and 9 alleles were detected for each of the locus. Together they have formed 151 haplotypes, with a diversity of 0.9937.

CONCLUSIONThe six STR loci included in this study were found to be highly polymorphic, and may provide useful markers for genetic analysis.

Alleles ; China ; Chromosomes, Human, Y ; Ethnic Groups ; genetics ; Gene Frequency ; Humans ; Korea ; ethnology ; Male ; Polymorphism, Genetic ; Tandem Repeat Sequences

OBJECTIVEX-linked spondyloepiphyseal dysplasia tarda (SEDL) is a rare osteochondrodysplasia caused by mutations of SEDL gene, which usually onset in late childhood without systemic complications. In this study, we have provided prenatal diagnosis for an affected family with a combined strategy including direct sequencing, fetal-sex identification and microsatellite linkage analysis.

METHODSTwo amniotic fluid samples from carrier gravida and 7 blood samples from individuals in this SEDL pedigree were obtained. Genomic DNA was extracted from the samples using standard phenol-chloroform method. SRY and AMEL genes were employed to assess fetal sex. Microsatellite DXS16 was genotyped for linkage analysis. A pathogenic mutation of the SEDL gene was identified by bi-directionally direct sequencing of the third exon as well as its exon/intron boundaries.

RESULTSTwo male fetuses were confirmed by fetal-sex assessment. The mutation of the SEDL gene was identified as a nucleotide substitution of the splice acceptor site in intron 2, IVS2-2A>C. DNA sequencing indicated that one fetus is hemizygote carrying the mutation, whilst another is not a carrier. Linkage analysis was identical with the sequencing results. Follow-up also confirmed the result of prenatal diagnosis.

CONCLUSIONFetal-sex assessment combined with microsatellite linkage analysis and bi-directionally direct sequencing is a more accurate and ready strategy for prenatal diagnosis of families affected with SEDL.

Genetic Diseases, X-Linked ; diagnosis ; Genetic Linkage ; Humans ; Male ; Osteochondrodysplasias ; diagnosis ; Prenatal Diagnosis ; Sequence Analysis, DNA

OBJECTIVETo study the influence of small interfering RNA (siRNA) targeting survivin on the biological behavior of bladder cancer.

METHODSOne pair of survivin target sequence-specific siRNA was designed, then siRNA/liposome complex was used to transfect bladder cancer cell line-T24. The efficiency of transfection and the apoptosis were detected by flow cytometry. The transcriptional level of survivin was analyzed using real time PCR. DNA sequence corresponding to siRNA targeting survivin was cloned into pRNAT-U6.1/Neo to produce plasmid targeting surviving.

RESULTSThe ratio of T24 cells releasing fluorescence in total cells were 92.3%; siRNA-survivin efficiently down-regulated survivin expression (mRNA) in a dose-and time-dependent manner. Its maximal effect was achieved at the concentration of 100 nmol/L, at which survivin expression level was down regulated by 75.91%. Similar results were found in the inhibition ratio of cell growth, which was 55.29%(P< 0.01). Simultaneously the apoptotic rate was markedly increased, which was 45.70%(P< 0.01). After cutting the vector with Bam H I and Hin d III and ligating the vector with the insert by using T4 ligase, the recombinant vector was confirmed by restriction digestion and DNA sequencing.

CONCLUSIONThe application of siRNA-survivin can markedly inhibit survivin expression in bladder cancer cell line, induce apoptosis and inhibit the growth of the tumor. It may be a new gene therapy tool for bladder cancer. The successful construction of the siRNA expressing plasmid will facilitate the application of RNA interference technique, and lay a foundation for further studies on the function of surviving.

Cell Line, Tumor ; Genetic Vectors ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Urinary Bladder Neoplasms ; genetics ; pathology

OBJECTIVETo investigate the influence of fluorescent protein expression on the proliferation of murine NIH3T3 cells, so as to provide a theoretical basis for cell tracing technology.

METHODSNIH3T3 cells were cultured in vitro, and were randomly divided into control, pLEGFP-N1 (with transfection of pLEGFP-N1 retroviral vector), pEGFP-N1 (with transfection of pEGFP-N1 vector) and pDsRed2-C1 (with transfection of pDsRed2-C1 vector) groups. Then the cells were screened by G418 for 3 weeks. The changes in cell adhesive rate were observed and the population doubling times was determined by growth curve.

RESULTSThere was obvious fluorescent protein expression in the transfected NIH3T3 cells after G418 selection, and the highest percentage of labeled NIH3T3 cells was found in pLEGFP-N1 group. The population doubling time in pDsRed2-C1 (40.3+/-0.7 h) , PEGFP-N1 (39.6 +/- 0.6 h) and pLEGFP-N1 (36.5 +/- 0.7 h) groups was evidently longer than that in control (27.9 +/- 0.6 h, P < 0.01), with high adhesive rate in each group.

CONCLUSIONThe expression of fluorescent protein exhibited some inhibitory effect on the proliferation of NIH3T3 cells in vitro. Since the inhibitory effect by retroviral vector was weaker compared with eukaryotic vector, it should be the first choice for fluorescent protein labeling during cell transplantation.

Animals ; Cell Culture Techniques ; Cell Proliferation ; Green Fluorescent Proteins ; biosynthesis ; Mice ; NIH 3T3 Cells ; Transfection

Objective: To understand the etiological characteristics of bacterial diarrhea in different areas, including large cities, mid-sized/small cities and rural area, in China. Methods: A cross-sectional surveillance was conducted in 17 provinces of China from 2010 to 2014. The acute diarrhea outpatients were selected from clinics or hospitals in large cities, mid-sized/small cities, including rural-urban fringe zones, and rural areas. The demographical and clinical characteristics of the patients were collected by using questionnaire, and stool samples were taken from them for laboratory detection of 17 kinds of bacteria. The differences in pathogen positive rates (PPR) and pathogen spectrum across the cases from three-type areas were compared. The different infection risk in different cases were analyzed with unconditional logistic regression model. Results: In our study, we enrolled 9 253 cases from large cities, 5 138 cases from rural areas and 13 683 cases from midsized/small cites. The pathogen with largest differences in infection rate across the three-type areas was Shigella (S.) flexneri (rural area: 5.81%, mid-sized/small city: 2.78%, large city: 0.46%), followed by Aeromonas (A.) hydrophila (rural area: 2.14%, mid-sized/small city: 0.96%, large city: 0.48%). Compared with cases in large cities, the cases in mid-sized/small cities and rural areas had higher infection risks for S. flexneri (mid-sized/small city: OR=6.481, 95%CI: 4.666-9.002, rural area: OR=11.304, 95%CI: 8.018-15.938) and A. hydrophila (mid-sized/small city: OR=1.992, 95%CI:1.401-2.832, rural area: OR=4.083, 95%CI: 2.833-5.884). The constituent ratio of diarrheagenic Escherichia coli and Salmonella increased with the urbanization development, while the ratios of Shigella and A. hydrophila had an opposite trend. S. sonnei (60.00%) was the predominant serogroup of Shigella in urban infections, while S. flexneri (77.37%) was the predominant serogroup in rural infections. Conclusion: The differences in pathogen spectrum of bacterial diarrhea were obvious across large cities, mid-sized/small cities and rural areas in China, especially the differences in the infection rates of S. flexneri and A. hydrophila.
Adolescent ; Adult ; Bacterial Infections/microbiology* ; Child ; China/epidemiology* ; Cross-Sectional Studies ; Diarrhea/microbiology* ; Dysentery/epidemiology* ; Escherichia coli/pathogenicity* ; Feces/virology* ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Rural Population ; Salmonella/pathogenicity* ; Shigella/pathogenicity* ; Suburban Population ; Urban Population

A 54-year-old asymptomatic man underwent a video-assisted thoracoscopic left pneumonectomy for squamous-cell carcinoma. During the surgery, a complete left pericardial defect was unexpectedly discovered, but no special intervention was made. The preoperative chest CT was reciewed, which showed the heart extended unusually to the left, but the left pericardial defect was not evident. The operation time was 204 min and the patient was discharged from hospital upon recovery 9 days after the surgery. The pathological result indicated moderately differentiated squamous-cell carcinoma (T2N1M0, stage ⅡB), and metastasis was found in the parabronchial lymph nodes (3/5). The patient did not receive chemotherapy after the surgery, and there was no signs of recurrence 6 months after the surgery. Complete pericardial defects usually do not endanger the lives of patients, and if the patient is asymptomatic, pneumonectomy is feasible.

Objective To investigate the significance of spread through air spaces (STAS) in early-stage non-small cell lung cancer (NSCLC) patients undergoing either sublobar resection or lobectomy by pooling evidence available, and to assess the accuracy of frozen sections in determining types of resection among patients with suspected presence of STAS. Methods Studies were identified by searching databases including PubMed, EMbase, Web of Science, and The Cochrane Library from inception to July 2022. Two researchers independently searched, screened, evaluated literature, and extracted data. Statistical analysis was conducted using RevMan 5.4 and STATA 15.0. The Newcastle-Ottawa Scale (NOS) was used to evaluate the quality of the study. Results A total of 26 studies involving 23 surgical related studies (12 266 patients) were included, among which, 11 compared the outcomes of lobectomy with sublobar resection in the STAS-positive patients. NOS score≥6 points. Meta-analysis indicated that presence of STAS shortened patients' survival in both lobectomy group and sublobar resection group (RFS: HR=2.27, 95%CI 1.96-2.63, P＜0.01; OS: HR=2.08, 95%CI 1.74-2.49, P＜0.01). Moreover, lobectomy brought additional survival benefits to STAS-positive patients compared with sublobar resection (RFS: HR=1.97, 95%CI 1.59-2.44, P＜0.01; OS: HR=1.91, 95%CI 1.47-2.48, P＜0.01). Four studies were included to assess the accuracy of identifying presence of STAS on intraoperative frozen sections, of which the pooled sensitivity reached 55% (95%CI 45%-64%), the pooled specificity reached 92% (95%CI 77%-97%), and the pooled area under the curve was 0.68 (95%CI 0.64-0.72) based on the data available. Conclusion This study confirms that presence of STAS is a critical risk factor for patients with early-stage NSCLC. Lobectomy should be recommended as the first choice when presence of STAS is identified on frozen sections, as lobectomy can prolong patients' survival compared with sublobar resection in STAS-positive disease. The specificity of identifying STAS on frozen sections seems to be satisfactory, which may be helpful in determining types of resection. However, more robust methods are urgently in need to make up for the limited sensitivity and accuracy of frozen sections.

OBJECTIVE: To study the anatomical characteristics of blood vessels in the lateral segment of the vertebral body through the surgical approach of oblique lumbar interbody fusion (OLIF) using MRI imaging, and evaluate its potential vascular safety zone. METHODS: The lumbar MRI data of 107 patients with low back and leg pain who met the selection criteria between October 2019 and November 2022 were retrospectively analyzed. The vascular emanation angles, vascular travel angles, and the length of vessels in the lateral segments of the left vertebral body of L ₁-L ₅, as well as the distance between the segmental vessels in different Moro junctions of the vertebral body and their distances from the edges of the vertebrae in the same sequence (bottom marked as I, top as S) were measured. The gap between the large abdominal vessels and the lateral vessels of the vertebral body was set as the lateral vascular safe zones of the lumbar spine, and the extent of the safe zones (namely the area between the vessels) was measured. The anterior 1/3 of the lumbar intervertebral disc was taken as the simulated puncture center, and the area with a diameter of 22 mm around it as the simulated channel area. The proportion of vessels in the channel was further counted. In addition, the proportions of segmental vessels at L ₅ without a clear travel and with an emanation angel less than 90° were calculated. RESULTS: Except for the differences in the vascular emanation angles between L ₄ and L ₅, the vascular travel angles between L ₁, L ₂ and L ₄, L ₅, and the length of vessels in the lateral segments of the vertebral body among L ₁-L ₄ were not significant ( P>0.05), the differences in the vascular emanation angles, vascular travel angles, and the length of vessels between the rest segments were all significant ( P<0.05). There was no significant difference in the distance between vessels of L ₁, L ₂ and L ₂, L ₃ at Moro Ⅰ-Ⅳ junctions ( P>0.05), in L ₃, L ₄ and L ₄, L ₅ at Ⅱ and Ⅲ junction ( P>0.05). There was no significant difference in the vascular distance of L ₂, L ₃ between Ⅱ, Ⅲ junction and Ⅲ, Ⅳ junction, and the vascular distance of L ₃, L ₄ between Ⅰ, Ⅱ junction and Ⅲ, Ⅳ junction ( P>0.05). The vascular distance of the other adjacent vertebral bodies was significant different between different Moro junctions ( P<0.05). Except that there was no significant difference in the distance between L ₂I and L ₃S at Ⅰ, Ⅱ junction, L ₃I and L ₄S at Ⅱ, Ⅲ junction, and L ₂I and L ₃S at Ⅲ, Ⅳ junction ( P>0.05), there was significant difference of the vascular distance between the bottom of one segment and the top of the next in the other segments ( P<0.05). Comparison between junctions: Except for the L ₃S between Ⅰ, Ⅱ junction and Ⅱ, Ⅲ junction, and L ₅S between Ⅰ, Ⅱ junction and Ⅱ, Ⅲ and Ⅲ, Ⅳ junctions had no significant difference ( P>0.05), there were significant differences in the distance between the other segmental vessels and the vertebral edge of the same sequence in different Moro junctions ( P<0.05). The overall proportion of vessels in the simulated channels was 40.19% (43/107), and the proportion of vessels in L ₁ (41.12%, 44/107) and L ₅ (18.69%, 20/107) was higher than that in the other segments. The proportion of vessels in the channel of Moro zone Ⅰ (46.73%, 50/107) and zone Ⅱ (32.71%, 35/107) was higher than that in the zone Ⅲ, while no segmental vessels in L ₁ and L ₂ were found in the channel of zone Ⅲ ( χ ²=74.950, P<0.001). Moreover, 26.17% (28/107) of the segmental vessels of lateral L ₅ showed no movement, and 27.10% (29/107) vascular emanation angles of lateral L ₅ were less than 90°. CONCLUSION L ₁ and L ₅ segmental vessels are most likely to be injured in Moro zones Ⅰ and Ⅱ, and the placement of OLIF channels in L _{4, 5} at Ⅲ, Ⅳ junction should be avoided. It is usually safe to place fixation pins at the vertebral body edge on the cephalic side of the intervertebral space, but it is safer to place them on the caudal side in L _{1, 2} (Ⅰ, Ⅱ junction), L _{3, 4} (Ⅲ, Ⅳ junction), and L _{4, 5} (Ⅱ, Ⅲ, Ⅳ junctions).
Humans ; Retrospective Studies ; Spinal Puncture ; Magnetic Resonance Imaging ; Anticoagulants ; Bone Nails

Objective To compare the efficacy and safety of different cyclin-dependent kinase 4/6 inhibitors (CDK4/6i) combined with endocrine therapy (ET) for the treatment of hormone receptor-positive (HR+)/human epidermal growth factor receptor 2-negative (HER2−) advanced or metastatic breast cancer. Methods Randomized controlled trials (RCTs) on CDK4/6i for the treatment of HR+/HER2− metastatic or advanced breast cancer were retrieved from databases including PubMed, EMbase, Web of Science, The Cochrane Library, CNKI, Wanfang, VIP, and SinoMed, with the search period ranging from database inception to August 2023. Bayesian network meta-analysis was conducted using R 4.2.0 software. Results A total of 18 RCTs from 25 articles, involving 8 031 patients and 11 treatment regimens, were included. There was no significant difference in progression-free survival (PFS) or overall survival (OS) among different CDK4/6i+ET combinations. The highest cumulative probability for PFS was observed with dalpiciclib (DAL)+fulvestrant (FUL), while ribociclib (RIB)+FUL ranked first for OS. In terms of efficacy, abemaciclib (ABE)+aromatase inhibitors (AI) and ABE+FUL ranked first in objective response rate and clinical benefit rate, respectively. Regarding safety, statistically significant difference in grade 3-4 adverse events was observed among certain types of CDK4/6i (P<0.05). Conclusion Current evidence suggests that CDK4/6i+ET is superior to ET alone for the treatment of HR+/HER2− advanced/metastatic breast cancer. Different CDK4/6i+ET combinations demonstrate comparable or similar efficacy; however, the incidence of adverse reactions is higher with combination therapy. Treatment regimens should be selected based on individual conditions.