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.

Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.

Objective To establish a quality control method for detecting impurities in chloral hydrate raw materials, improve the quality standards and control limits of raw materials. Methods The determination methods of chloroform and halogenated carboxylic acid in chloral hydrate were established to monitor the change of impurities in chloral hydrate through stability. Results The research and establishment of chloroform and halogenated carboxylic acid methods met the requirements of relevant regulations for analytical methodology verification, which could accurately detect four impurities in raw materials and preparations by one method. Conclusion The study provides technical support for the improvement and optimization of the quality standards of chloral hydrate and preparations. It is very necessary to implement the impurity monitoring in preparation research and production process by the chloral hydrate impurity detection and the stability comparison of this product at high temperature and light, which could largely promote the safety of medication.

OBJECTIVE: To carry out optical genome mapping (OGM) for a Chinese pedigree with a rare paracentric reverse insertion of chromosome 17. METHODS: A high-risk pregnant woman identified at the Prenatal Diagnosis Center of Hangzhou Women's Hospital in October 2021 and her family members were selected as the study subjects. Chromosome G banding analysis, fluorescence in situ hybridization (FISH), single nucleotide polymorphism array (SNP array) and OGM were applied to verify the balanced structural abnormality of chromosome 17 in the pedigree. RESULTS: Chromosomal karyotyping analysis and SNP array assay have identified a duplication of 17q23q25 in the fetus. Karyotyping analysis of the pregnant woman showed that the structure of chromosome 17 was abnormal, whilst SNP array has detected no abnormality. OGM revealed that the woman has carried a paracentric reverse insertion, which was confirmed by FISH. The karyotype of her husband was normal. CONCLUSION The duplication of 17q23q25 in the fetus has derived from a paracentric reverse insertion of chromosome 17 in its mother. OGM has the advantage for delineating balanced chromosome structural abnormalities.
Pregnancy ; Humans ; Female ; Pedigree ; In Situ Hybridization, Fluorescence ; Chromosomes, Human, Pair 17/genetics* ; East Asian People ; Chromosome Aberrations ; Prenatal Diagnosis ; Chromosome Mapping ; Chromosome Inversion

Objective: To access the effect of maternal cell contamination (MCC) on the detection of copy number variation (CNV) by chromosome microarray analysis (CMA) in prenatal diagnostic samples. Methods: Amniotic fluid DNA samples were collected from Department of Prenatal Diagnosis (Screening) Centre of Hangzhou Women′s Hospital from December 2016 to August 2018. Copy number variations (CNVs) were identified in these DNA samples by CMA and normal female genomic DNA was added to simulate different proportions of maternal cells contamination. The simulated samples were tested using an Agilent microarray chromosome chip 180K CGH (Agilent 180K CGH) and the results were analyzed by Agilent CytoGenomics software. Results: The results showed that duplications of CNV could not be detected at>38.4% MCC. Deletions of CNV could not be detected at>41.3% of MCC. With the increase of MCC ratio, the detection rate of CNV decreased gradually. The array and software allowed detection of longer copy-number variations at higher levels of maternal cell contamination than shorter copy-number variations. For the same size CNV, the detection ability of the deleted CNV was slightly higher than that of duplication. In the male sample, the observable shift of the X/Y chromosome ratio at>10% MCC can be detected by the microarray. Conclusions When the MCC is higher than a particular ratio, it can affect the CMA detection of CNV. Based on the Agilent 180K CGH detection results for MCC mimics and the CNV detection specificity principle, the MCC threshold of the Agilent 180K CGH is set to 30% in view of conservative principles.

Objective To evaluate the clinical value of percutaneous AngioJet thrombectomy for acute iliofemoral deep venous thrombosis.Methods 50 patients were divided into AngioJet thrombectomy group(25 cases) compared with bolus injection of urokinase through catheter in control group (25 cases).When the thrombosis was completely dissolved,the PTA and stents were implanted in those patients presenting May-Thurner syndrome in both groups.Inferior vena cava filter was retrieved from the patients and the thrombolytic therapy was discontinued.The patency rate of iliofemoral venous was assessed by CTA at 1 and 6 months after the procedure.Results The average dosage of urokinase in AngioJet patients was (880 000 ± 354 000) units.The mean duration time of catheter-directed thrombolysis was (42.2 ± 16.7) hours and the average hospitalization time is (3.8 ± 0.8) days.Grade Ⅲ thrombolysis was achieved in 18 limbs and grade Ⅱ lysis in 6 limbs,grade Ⅰ lysis in 1 limb.The mean duration time of catheter-directed thrombolysis was (129.6 ± 32.2) hours.The average dose of urokinase was (4 100 000 ± 1 060 000) units and the average hospitalization time was (7.9 ± 1.4) days in control group.Grade Ⅲ thrombolysis was achieved in 10 limbs,grade Ⅱ lysis in 9 limbs and grade Ⅰ lysis in 6 limbs.The dosage of urokinase,duration time of catheter-directed thrombolysis,time in hospital and clinical effects were statistically different between the two groups (all P ＜ 0.05).There were no recurrent thrombosis in all the patients after 6 months of treatment.Venous patency was maintained in all patients with implanted stent.Conclusions Percutaneous AngioJet thrombectomy with adjunctive thrombolytic therapy is an effective,safe treatment modality in patients with acute iliofemoral vein thrombosis.

Objective To determine the dose-response relationship of sufentanil blunting responses to double-lumen endotracheal intubation when combined with propofol given by target-controlled infusion (TCI) in patients with pulmonary tuberculosis.Methods One hundred patients of both sexes with pulmonary tuberculosis,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 24-58 yr,with body mass index ＜30 kg/m2,with Mallampati grade Ⅰ or Ⅱ,undergoing thoracic surgery under general anesthesia,were divided into Ⅰ-Ⅴ groups (n =20 each) using a random number table.Anesthesia was induced with iv sufentanil 0.35 μg/kg (group Ⅰ),0.40 μg/kg (group Ⅱ),0.45 μg/kg (group Ⅲ),0.50 μg/kg (group Ⅳ) and 0.55 μg/kg (group Ⅴ),and propofol TCI (target plasma concentration 3.5 μg/ml) and iv vecuronium 0.15 mg/kg.The patients were endotracheally intubated and mechanically ventilated.The response to double-lumen endotracheal intubation was defined as positive when mean arterial pressure increased by＞ 20％ of the baseline value and/or heart rate ＞ 90 bpm within 5 min after intubation.The median effective dose (ED50),ED95 and 95％ confidence interval (95％ CI) of sufentanil blunting the responses to double-lumen endotracheal intubation were calculated by Probit analysis.Results The ED50 (95％ CI) and ED95 (95％ CI) of sufentanil blunting the responses to intubation were 0.411 (0.370-0.441) μg/kg and 0.635 (0.556-0.888) μg/kg,respectively,when combined with propofol given by TCI.Conclusion When combined with propofol given by TCI (target plasma concentration 3.5 μg/ml),the ED50 and ED95 of sufentanil blunting the responses to double-lumen endotracheal intubation are 0.411 and 0.635 μg/kg,respectively,in patients with pulmonary tuberculosis.

Objective To discuss the clinical application of mechanical thrombectomy with AngioJet system for acute lower limb arterial ischemia (ALI).Methods A total of 12 ALI patients,who underwent percutaneous mechanical thrombectomy with AngioJet systemn during the period from January 2015 to November 2016,were enrolled in this study.The clinical data were retrospectively analyzed.The blood flow classification score after thrombolysis in myocardial infarction (TIMI) was used to evaluate the blood perfusion condition,and Cooley standard of efficacy score was used to assess the clinical curative effect.Results The technical success rate of mechanical thrombectomy with AngioJet system was 91.7％ (11/12).The average restored perfusion time was (1.5±0.6) hours.The clinical success rate and limb salvage rate were 83.3％ (10/ 12) and 91.7％ (11/12),respectively.The TIMI flow scores were improved from preoperative grade 0 (n=8) and Ⅰ (n=4) to postoperative grade 0 (n=1),Ⅰ (n=3) and Ⅱ (n=8).In 11 patients (91.7％) the symptoms of lower limb arterial ischemia were strikingly improved after mechanical thrombectomy.In one patient,the postoperative TIMI flow score remained 0 as preoperative state and the symptoms of lower limb arterial ischemia were not improved although catheter directed thrombolysis therapy was employed for 24 hours,and above-knee surgical amputation had to be carried out.Cooley efficacy score showed that complete cure was seen in 4 patients (33.3％),good response in 6 patients (50.0％),general improvement in one patient (8.3％) and pool response in one patient (8.3％).No severe bleeding complications occurred.Conclusion Percutaneous mechanical thrombectomy with AngioJet can rapidly recover the blood perfusion in patients with ALI,thus,further deterioration of the disease can be prevented and the limb salvage rate can be improved.Therefore,this technique has good clinical application value.

Objective To compare the dose, clinical efficacy and acute adverse reactions of intensity modulated radiotherapy(IMRT)and three-dimensional conformal radiotherapy(3D-CRT)combined with three-dimensional brachytherapy (3D-BT) in the treatment of concurrent radiotherapy and chemotherapy for advanced stage cervical cancer patients. Methods Data collection was performed from January 2011 to November 2015 in Chinese PLA General Hospital and Inner Mongolia Cancer Hospital.All 89 patients with advanced stage (Ⅱb-Ⅲb) cervical cancer were treated by pelvic radiotherapy and concurrent chemotherapy, 46 cases of them received IMRT and 3D-BT(IMRT group), 43 cases received 3D-CRT and 3D-BT(3D-CRT group),along with cisplatin chemotherapy.The dose accumulation of external beam radiotherapy and 3D-BT was calculated by deformable image registration to analyze clinical efficacy, acute adverse reactions and prognosis of the two groups.Results (1)Dose of radiotherapy:planning target volume(PTV)coverage of IMRT group and 3D-CRT group were respectively(95.4±4.7)% and(95.1±5.1)%, without significant differences (t=0.289, P=0.773). Compared with the patients treated with 3D-CRT, the volumn receiving at least 30 Gy (V30), V50of rectum, colon, bladder and small intestine and V20of bone marrow in the IMRT group were significantly decreased (P<0.05). Regarding the combined dose, the maximum dose (Dmax) and the minimum dose received by the most exposed 2 cm3volume of the analyzed organ(D2CC)of rectum,colon,bladder and small intestine of IMRT group were significantly lower than those of 3D-CRT group (P<0.05). (2) Short-term efficacy: the effective rate of IMRT and 3D-CRT group were respectively 93% (43/46) and 91% (39/43), with no significant differences (χ2=0.237,P=0.626). (3) Acute adverse reactions: compared with 3D-CRT, IMRT could significantly reduce grade 1-2 acute toxicity in gastrointestinal [63%(29/46) vs 84%(36/43)], genitourinary [17%(8/46) vs 37%(16/43)] and hematologic [57%(26/46) vs 79%(34/43)] system (all P<0.05). There were no significant differences of grade 3 acute adverse reactions of gastrointestinal,genitourinary and hematologic system between two groups(all P>0.05). No grade 4 acute adverse reactions were observed. (4) Prognosis: the overall survival rate at 1, 2-year of IMRT and 3D-CRT group were respectively 95.6%,89.1% and 93.1%,86.1%.The progression-free survival rateat 1, 2-year of IMRT and 3D-CRT group were 91.1%, 89.1% and 88.4%, 86.1%, respectively. There were no significant differences in overall survival rate and progression-free survival rate between two groups (P>0.05). Conclusions Compared with 3D-CRT, IMRT combined with 3D-BT has dosimetry advantages based on dose accumulation algorithms by deformable image registration. IMRT could ensure clinical efficacy and significantly reduce the incidence rate of acute toxicities.

Objective To assess the safety and efficacy of transcatheter thrombolysis combined with percutaneous puncture balloon dilatation for acute occlusion of forearm arteriovenous fistulas.Methods A total of 20 cases with acute thrombosed arteriovenous fistulas were treated with transcatheter thrombolysis therapy by distal puncture of the brachial artery,and the patients with arteriovenous fistulas stenosis more than 50％ were conducted percutaneous puncture balloon dilatation.Results Acute thrombosed arteriovenous fistulas was dredged in all 20 patients by transcatheter thrombolysis combined with percutaneous puncture balloon dilatation,without serious complications such as hemorrhage,pulmonary embolism and others.After follow-up for 3 to 6 months,16 cases maintained smooth,recurrent thrombosis was founded in 4 cases,among whom 2 were reopened after transcatheter thrombolysis in combination and percutaneous puncture balloon dilatation and 2 failed to dredged.Conclusion Transcatheter thrombolysis in combination with percutaneous puncture balloon dilatation is a safe and effective treatment for patients with acute thrombosed arteriovenous fistulas.