BACKGROUND:Osteoporotic fracture is the most serious complication of osteoporosis.Previous studies have demonstrated that gut microbiota has a regulatory effect on skeletal tissue and that gut microbiota has an important relationship with osteoporotic fracture,but the causal relationship between the two is unclear. OBJECTIVE:To explore the causal relationship between gut microbiota and osteoporotic fractures using Mendelian randomization method. METHODS:The genome-wide association study(GWAS)datasets of gut microbiota and osteoporotic fracture were obtained from the IEU Open GWAS database and the Finnish database R9,respectively.Using gut microbiota as the exposure factor and osteoporotic fracture as the outcome variable,Mendelian randomization analyses with random-effects inverse variance weighted,MR-Egger regression,weighted median,simple model,and weighted model methods were performed to assess whether there is a causal relationship between gut microbiota and osteoporotic fracture.Sensitivity analyses were performed to test the reliability and robustness of the results.Reverse Mendelian randomization analyses were performed to further validate the causal relationship identified in the forward Mendelian randomization analyses. RESULTS AND CONCLUSION:The results of this Mendelian randomization analysis indicated a causal relationship between gut microbiota and osteoporotic fracture.Elevated abundance of Actinomycetales[odds ratio(OR)=1.562,95%confidence interval(CI):1.027-2.375,P=0.037),Actinomycetaceae(OR=1.561,95%CI:1.027-2.374,P=0.037),Actinomyces(OR=1.544,95%CI:1.130-2.110,P=0.006),Butyricicoccus(OR=1.781,95%CI:1.194-2.657,P=0.005),Coprococcus 2(OR=1.550,95%CI:1.068-2.251,P=0.021),Family ⅩⅢ UCG-001(OR=1.473,95%CI:1.001-2.168,P=0.049),Methanobrevibacter(OR=1.274,95%CI:1.001-1.621,P=0.049),and Roseburia(OR=1.429,95%CI:1.015-2.013,P=0.041)would increase the risk of osteoporotic fractures in patients.Elevated abundance of Bacteroidia(OR=0.660,95%CI:0.455-0.959,P=0.029),Bacteroidales(OR=0.660,95%CI:0.455-0.959,P=0.029),Christensenellacea(OR=0.725,95%CI:0.529-0.995,P=0.047),Ruminococcaceae(OR=0.643,95%CI:0.443-0.933,P=0.020),Enterorhabdus(OR=0.558,95%CI:0.395-0.788,P=0.001),Eubacterium rectale group(OR=0.631,95%CI:0.435-0.916,P=0.016),Lachnospiraceae UCG008(OR=0.738,95%CI:0.546-0.998,P=0.048),and Ruminiclostridium 9(OR=0.492,95%CI:0.324-0.746,P=0.001)would reduce the risk of osteoporotic fractures in patients.We identified 16 gut microbiota associated with osteoporotic fracture by the Mendelian randomization method.That is,using gut microbiota as the exposure factor and osteoporotic fracture as the outcome variable,eight gut microbiota showed positive causal associations with osteoporotic fracture and another eight gut microbiota showed negative causal associations with osteoporotic fracture.The results of this study not only identify new biomarkers for the early prediction of osteoporotic fracture and potential therapeutic targets in clinical practice,but also provide an experimental basis and theoretical basis for the study of improving the occurrence and prognosis of osteoporotic fracture through gut microbiota in bone tissue engineering.

Objective To investigate the prevalence of Cryptosporidium infection and the distribution of parasite species and genotypes among HIV-positive individuals in Jiangxi Province. Methods HIV-positive individuals' sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023. Subjects' stool samples were collected, and genomic DNA was extracted from stool samples. Nested PCR assay was performed based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium, and Cryptosporidium gp60 gene was amplified in stool samples positive for the SSU rRNA gene. The second-round PCR amplification product was checked with 1.5% agarose gel electrophoresis, and the products of suspected positive amplifications were sequenced, followed by sequence alignment. The phylogenetic tree was created using the Neighbor-Joining method with the software MEGA 11.0, to characterize the species, genotypes and sub-genotypes of Cryptosporidium. Results A total of 382 HIV-positive individuals were enrolled, with two cases identified with Cryptosporidium infection (0.52% prevalence), and both cases had no abdominal pain or diarrhea. Following sequencing and sequence alignment, the gene sequences of these two Cryptosporidium isolates shared 99.76% and 99.88% similarity with the gene sequence of C. meleagridis isolates. Phylogenetic analysis based on the Cryptosporidium SSU rRNA gene sequence identified the species of these two Cryptosporidium-positive stool samples as C. meleagridis. Following nested PCR amplification of the Cryptosporidium gp60 gene, sequencing and sequence alignment, the two C. meleagridis isolates were characterized as III eA17G2R1 and III bA25G1R1a sub-genotypes, and the sub-genotype III bA25G1R1a was firstly described in humans. Conclusion The prevalence of Cryptosporidium is low among HIV-positive individuals in Jiangxi Province. The likelihood of Cryptosporidium infection cannot be neglected among HIV-positive individuals without diarrhea.

Objective This paper investigates the effect of a multi-physical field fusion intervention based on transcranial direct current stimulation(tDCS),transcranial magnetic stimulation(TMS)and music on mental fatigue.Methods After fatigue induction,10 subjects received blank group stimulation,music group stimulation and tDCS-TMS-Music stimulation,respectively.tDCS stimulation sites were located in the bilateral frontal regions of the subjects,and TMS stimulation sites were located in the bilateral occipital regions of the hindbrain.Heart rate variability and reaction performance were measured before and after each intervention to determine the elimination effect of different intervention programs on mental fatigue.Results Compared with the control group,the tDCS-TMS-Music group showed significantly greater improvements in subjective mental workload,response performance,and heart rate variability.Conclusion The results of this study support that tDCS-TMS-Music can effectively alleviate mental fatigue induced by long-term cognitive performance tasks,and the intervention effect is better than music intervention and resting-state relief at the same time.

Objective:To discuss the effect of downregulating microRNA-208a(miR-208a)on the resistance of the colorectal cancer cells to 5-fluorouracil(5-FU),and to clarify its related molecular mechanism.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-208a and secreted frizzled-related protein 1(SFRP1)mRNA in the 5-FU-resistant colorectal cancer cell line HT-29/5-FU and its parent HT-29 cells.The HT-29/5-FU cells were transfected with miR-208a inhibitor plasmid and its negative control plasmid(inhibitor-NC),and SFRP1 small interfering RNA(si-SFRP1)and its negative control plasmid(si-NC),either separately or in combination,followed by treatment with 5-FU.The cells were divided into inhibitor-NC group,miR-208a inhibitor group,miR-208a inhibitor+si-NC group,and miR-208a inhibitor+si-SFRP1 group.MTT assay was used to detect the proliferation activities of the cells and the resistance indexes were calculated;Annexin V-FITC/PI double staining and flow cytometry were used to detect the apoptotic rates of the cells after treated with different concentrations of 5-FU;Western blotting method was used to detect the expression levels of SFRP1,β-catenin,P-glycoprotein(P-gp),and ATP-binding cassette subfamily B member 1(ABCB1)proteins in the cells in various groups;dual-luciferase reporter gene assay was used to validate the targeting relationship between miR-208a and SFRP1.Results:Compared with HT-29 cells,the expression level of miR-208a in the HT-29/5-FU cells was increased(P＜0.05),and the expression level of SFRP1 mRNA was decreased(P＜0.05).Compared with inhibitor-NC group,the proliferation activity of the cells in miR-208a inhibitor group was decreased(P＜0.05),the resistance index was decreased,the apoptotic rate was increased(P＜0.05),and the expression levels of β-catenin,P-gp,and ABCB1 proteins in the cells were decreased(P＜0.05).The dual-luciferase reporter gene assay results showed that SFRP1 was a target gene of miR-208a and miR-208a could negatively regulate the expression of SFRP1.Compared with miR-208a inhibitor+si-NC group,the proliferation activity of the cells in miR-208a inhibitor+si-SFRP1 group was increased(P＜0.05),the resistance index was increased,the apoptotic rate was decreased(P＜0.05),and the expression levels of β-catenin,P-gp,and ABCB1 proteins in the cells were increased(P＜0.05).Conclusion:Downregulation of miR-208a can improve the resistance of the HT-29/5-FU cells to 5-FU by targeting and upregulating the SFRP1 expression,thereby inhibiting the transmission of the Wnt signaling pathway.

Objective: To observe an abnormal expression of humoral immune response induced by memory B cells in tonsils and peripheral blood of patients with IgA nephropathy ( IgAN) , the variation of memory B cells after tonsillectomy , and to discover the role of tonsillectomy in IgAN .Methods: In the study , 28 patients were diagnosed as IgAN via renal biopsy , and 27 patients suffering from chronic ton-sillitis without nephritis and 10 normal human beings were selected as controls .The expression of memory B cells in the tonsils and peripheral blood was tested by flow cytometry , and the same method was used to test the variation of the expression of memory B cells in peripheral blood of patients with IgAN after tonsil -lectomy.Results:In this study , higher percentages of memory B cells were observed in tonsil and pe-ripheral blood of IgAN patients, which were 5.72%±5.26%, 4.92%±5.10%.After tonsillectomy, the percentage of memory B cells was 1 .10%±0 .65%, lower than that before tonsillectomy ( P <0.05).Meanwhile, in tonsils and peripheral blood , the percentage of memory B cells varied with the variation of the urinary findings of the IgAN patients .Conclusion:The percentage of memory B cell in tonsils and peripheral blood could predict disease progression of IgAN to a certain extent .

Objective To investigate the value of three-dimensional visualization system in the treatment of hilar cholangiocarcinoma.Methods The clinical data of 10 patients with hilar cholangiocarcinoma who were admitted to Chenggong Hospital of Xiamen University from January 2012 to September 2012 were retrospectively analyzed.The two-dimensional computed tomography images were converted to three-dimensional images with the three-dimensional visualization system,and then the volume of liver and tumor size,volume of liver to be resected,remnant liver volume were measured.Surgical procedure was planned based on the three-dimensional images,and the difference between the actual and planned surgical procedures was analyzed.The correlation between actual liver resection volume and predicted liver resection volume was analyzed by calculating the Pearson correlation coefficient.Results The mean liver volume,tumor size,predicted liver resection volume and remnant liver volume of the 10 patients detected by the three-dimensional visualization system were (1496 ± 162) ml,(67 ± 18) ml,(335 ± 241)ml and (1140 ± 197)ml,respectively.The average error rate of predicted liver resection volume was 6.4％.Surgical plan was made in accordance with the principle of curative hepatectomy,including 4 cases of left semihepatectomy,2 cases of right semihepatectomy,3 cases of partial liver resection and 1 case of palliative liver resection.The coincidence rate between the planned and actual surgical procedures was 9/10.R0 resection was performed on 7 patients,R1 resection on 1 patient and palliative resection on 2 patients.One patient received restrictive portal vein arterialization.Preoperative evaluation of the anatomy of blood vessels,bile ducts and tumors based on three-dimensional images was confirmed with operative findings.The accuracy of tumor typing by the three-dimensional visualization system was 8/10.The actual liver resection volume was (325 ± 258) ml,which was positively correlated with the predicted liver resection volume (r =0.902,P ＜ 0.05).Conclusion The three-dimensional visualization system is helpful in the treatment of hilar cholangiocarcinoma.

Objective To compare the dosimetric difference on organ at risk (OAR) in intensitymodulated radiotherapy (IMRT) with three-dimensional conformal radiotherapy (3D-CRT) in treatment with cervical carcinoma. Methods 36 cases of cervical cancer patients accepted radiotherapy both on IMRT and 3D-CRT in the same time,clinical target volume (CTV) focused on primary tumor and its lymphatic drainage area.Lymphatic drainage area was determined as follows:1.0 cm extension were added to the margin of pelvic vessels,the obturator lymph nodes region was extended by 1.8 cm from the pelvis inner margin.CTV does not include pelvic tissue. Data of dose distribution and irradiation volume of OAR on the basis of distributing on 45 Gy and 50 Gy were analyzed individually. The margins of planning target volume (PTV) was obtained based on CTV margins by which added 1.0 cm extension in inferior and superior directions,0.7 cm were added in all other directions. Results With the comparison of variety on irradiation volume of organs at risk (OAR) by DVH in both groups,underlying the treatment dose 45 Gy,the individual volume of D30,D40 and D45 were all lesser in IMRT plan than that of 3D-CRT plan. Even on the dose 50 Gy at pelvic wall lymph nodes regions,the volume of OAR was lesser than that in 3D-CRT plan.Conclusion In the radiotherapy of cervical cancer,IMRT should have improved protections for tumor surrounding OAR which has domestic superiority in clinical application.

Objective To analyze the significance of postoperative radiotherapy for lymph node positive patients after radical resection of esophageal carcinoma.Methods Two hundred and sixty patients with esophageal squamous cell cancer,aged ≤70,with the performance status score of0 -1,who had undergone radical resection were divided into 2 equal groups:surgery alone group (Group A ) and surgery plus radiotherapy group (Group B).Group A was classified into 3 sub-groups:Group A1 (n =42)without lymph node involvement,Group A2 (n =43 ) with 1 to 3 involved lymph nodes,and Group A3(n =45) with ≥4 involved lymph nodes.Group B was classified into 3 sub-groups:Group B1 (n =43 )without lymph node involvement,Group B2 (n =44) with 1 to 3 involved lymph nodes,and Group B3(n =43 ) with ≥4 involved lymph nodes.The patients were followed up till death.Results The 1-,3-,and 5-year overall survival rates of Group A were 71.5％,35.4％ and 20％,respectively,all significantly lower than those in Group B (76.2％,48.5％ and 36.2％,respectively,x2 =7.822,P ＜0.05).The 1-,3-,and 5-year survival rates of Groups A1 were 83.3％,52.3％,and 38.1％,respectively,all not significantly different from those of Group B1 (81.3％,58.1％,and 46.5％,respectively,x2 =0.283,P ＞ 0.05 ).The 1-,3-,and 5-year survival rates of Groups A2 were 69.8％,34.9％,and 18.6％,respectively,all significantly lower than those of Group B2 (77.3％,47.7％,and 40.9％,respectively,x2 =4.188,P ＜ 0.05).The 1-,3-,and 5-year survival rates of Groups A3 were 62.2％,20％,and 4.4％,respectively,all significantly lower than those of Group B3 ( 69.8％,39.5％,and 20.9％,respectively,x2 =6.168,P ＜ 0.05).The 5-year metastatic lymph node rates of Groups A1 to A3 were 30.9％,53.4％,and 66.7％,respectively,all significantly higher than those of Groups B1 to B3 ( 11.6％,22.7％,and 30.2％,respectively,x2 =4.753,8.741,and 11.682,respectively,all P ＜0.05).The 5-year distant metastasis rates of Groups A1 to A3 were 11.9％,20.9％,and 31.1％,respectively,all not significantly different from those of Groups B1 to B3 (13.9％,20.4％,and 25.6％,respectively,x2 =0.079,0.003,and 0.203,respectively,all P ＞ 0.05 ).Conclusions Postoperative radiotherapy increases the survival rate of lymph node positive patients,but shows little efficacy on the lymph node negative patients.It reduces the occurrence of lymph node metastasis,even in the lymph node negative patients,and does not increase the morbidity of complications,especially that of anastomotic stenosis.The number of metastatic lymph node is one of the important factors affecting the survival of esophageal carcinoma.Distant metastasis increases along with the number of metastatic lymph nodes.

Objective To evaluate the feasibility , toxicity and clinical efficacy of intensity-modulated radiotherapy using the simultaneous integrated boost (SIB- IMRT) and concurrent chemotherapy for advanced nasopharyngeal carcinoma. Methods Thirty nsopharyngeal carcinoma were treated with full course IMRT including nasopharynx and full neck to supraclavicle. The radiotherapy dosage is 68 Gy to the target. Concurrent chemotherapy was given, and the regimen was DDP 40 mg/m2/weekly.Results The mean dose of covering gross tumor volume(PGTV) (D95) in the nasopharynx was 70.48 Gy, and the mean volume of PGTV1 receiving the 95 % dose(V95) was 98. 46 %. The mean dose of PGTV1, PGTV2, PCTV1 and PCTV2 in the targets were 70.8 Gy, 66.4 Gy, 62.3 Gy and 54.8 Gy. According to the evaluation, the acute skin,mucositis and salivary toxicity with grade Ⅲ in those patients were 3.3 %, 10 %, 6.6 %. The patients developed different blood toxicity, but didn't affect their treatment. The median follow-up time was 6.5 months, and disease free survival rate was 100 %. Conclusion SIB-IMRT yields well dose distribution and acceptable toxicity in advanced stage nasopharyngeal carcinoma. The preliminary clinical result is encouraging.

Objective To investigate the clinicopathological difference and prognosis of colorectal adenocarcinomas including mutinous, Signet-ring cell, papillary and tubular carcinomas. Methods Two thousand and eighty-nine patients with colorectal cancer underwent colorectal operation between August 1994 and April 2007. The clinicopathological characteristics of mucinous adenocarcinoma (n=144), signet-ring cell carcinoma (n=25), papillary and tubular carcinomas (n= 1837) were compared expect of other types of cancer (n = 83). The single factor and Logistic regression methods were used to analyze the clinicopathological parameters that influence the prognosis of colorectal cancer such as age, location of the tumor, staging, peritoneum and pathological typing. The survival rates of patients with above three types of adenocareinomas were analyzed. Results The mean age of onset was lowest in patients with mutinous adenocarcinomas [(54. 20 ± 16.25) years] compared with that in patients with signet-ring cell cancer [(40.43 ± 12.88)years] or papillary and tubular carcinomas [(58. 73 ±13.62)]. There were significant differences in gender, size and location of the tumor, TNM staging, peritoneal metastasis, lymph node involvement and adjacent organ invasion among three groups (all P values <0.05). The single factor and Logistic regression analysis revealed that both mucinous adenocarcinoma and signet-ring cell carcinoma were risk factors ot prognosis. The patients with mucinous adenocarcinoma or signet-ring cell tumor were poor in long-term overall survival in comparison with patients with papillary and tubular carcinoma (P<0. 001). Conclusions The colorectal mucinous and signet-ring cell adenocarcinomas are risk factors for prognosis of colorectal cancer, which imply the poor outcome.