Objective To construct the recombinant plasmid containing human filaggrin gene,purify and identify the immunoreactivity of the recombinant protein,and establish the indirect ELISA to detect AFA for diagnosis of RA.Methods The constructed plasmids were transformed into E. Coli Rosettagami(DE3).This fusion protein was purified by NAT chromatography.ELISA coated with the fusion protein Was established to detect the AFA in serum of patients,which included 114 cases of RA,56 cases of SLE,32 cases of OA and 40 cases of normal controls. The correlation between the results of AFA and anti-CCP in RA group were compared. Results 321 bp fragment of filaggrin gene was amplified and the recombinant expression vector pET-28a( + )-filaggrin was constructed. The sequence of filaggrin gene was the same as the sequence reported in the literatures. The Rosetta-gami (DE3) strains of E. Coli with recombinant vector showed high level of filaggrin protein after induction. The SDS-PAGE showed that the plasmid expressed the filaggrin fusion protein with molecule weight of 14 000 Da. The expression protein could be purified by Ni-NAT with activity. The absorbance value of AFA in RA group was 0.473 ±0. 248 while they were 0. 160 0. 088, 0. 121±0. 070, 0.050 0. ±018 in SLE, OA and normal groups respectively. There were significant differences of absorbance values of AFA between RA and SLE, OA, control group (t = 12.004, 14. 464, 18.078, P<0. 01, respectively). The positivities of anti-filaggrin in RA, SLE and OA were 48.2%, 5.4% and 3. 1% respectively. The positivities of AFA were significantly different between RA, OA and normal control groups (x~2 = 67. 088, P < 0. 01). There was positive correlation of results between AFA and anti-CCP antibody (r = 0.42, P < 0. 05 ) . The consistency rate of results between AFA and anti-CCP was 70. 1%. Anti-CCP was negative in 10 out of 114 patients with AFA positive. AFA can be used to diagnose RA with sensitivity of 48. 2% , specificity of 96.9% , positive predictive value of 93. 2% and negative predictive values of 67. 9% . Conclusions The purified human filaggrin fusion protein is successfully purified. The indirect ELISA method based on the recombinant protein shows good sensitivity and specificity. Joint detection with AFA and anti-CCP can improve the positive rate of detection.

Objective To investigate the clinical outcomes derived from Ningmitai combined with tamsulosin to prevent double-J stent syndrome after laser lithotripsy with ureteroscope. Methods 117 patients underwent laser lithotripsy with ureteroscope and then placed a double-J stent for draining were collected from January 2010 to January 2013. Patients with double-J stent placement were divided into four groups determined by dosage regimen. Tamsulosin group (30 cases) was treated with tamsulosin (0.4 mg once daily) lonely, Ningmitai group (29 cases) was treated with Ningmitai (1.52 g, trice time a day) lonely, tamsulosin combined Ningmitai group (30 cases) was treated with tamsulosin and Ningmitai at the same time, operation control group (28 cases) was neither tamsulosin nor Ningmitai. The catheter was removed on the 3rd day post-lithotripsy and then remained double-J stent for 1 month. The scores of urinary tract, pain and the incidence of gross hematuria were assessed. Results The significant differences in the improvement of symptom score (χ2=22.038, P=0.000), pain score (χ2=9.876, P=0.020) and hematuria (χ2=8.000, P=0.046) were found among tamsulosin group, Ningmitai group, and tamsulosin combined Ningmitai group. The number of patients with symptomless, slight symptom in tamsulosin combined Ningmitai group were higher than those of tamsulosin group, Ningmitai group, operation control group (symptomeless:14 vs. 6, 3 and 2 cases;slight symptom:13 vs. 9, 5, 4 cases). The number of patients with>Ⅱpain score (7 vs. 9, 14, 17 cases) and incidence of hematuriag [26.6%(8/30) vs. 56.7%(17/30), 58.6% (17/29), 53.6% (15/28)] were lower in tamsulosin combined Ningmitai group than those of tamsulosin group, Ningmitai group, operation control group. The drug combination of Ningmitai with tamsulosin had the synergism to relived symptom and pain, and showed the more obviousthan lonely use. Conclusion The drug combination of Ningmitai with tamsulosin can be used in clinic for prophylactic purpose to prevent double-J syndrome.

Objective: To investigate the clinical efficacy of minimally invasive plate oseoynthesis (MIPO) assisted by a self-designed lower limb axial tractor in the treatment of complex tibial plateau fractures. Methods: The data were retrospectively analyzed of the 31 patients who had undergone surgical treatment for complex tibial plateau fractures from June 2015 to April 2018 at Department of Orthopeadics, Hospital Affiliated to Binzhou Medical College. MIPO was conducted with the assistance of the axial tractor in 17 of them. They were 12 men and 5 women with an age of 50.8±14.7 years, with 8 cases of Schatzker type Ⅴ and 9 ones of Schatzker type Ⅵ. MIPO was conducted without the assistance of the axial tractor but with the assistance of manipulation reduction in the other 14 patients. They were 9 men and 5 women with an age of 48.9±9.5 years, with 6 cases of Schatzker type Ⅴ and 8 ones of Schatzker type Ⅵ. The 2 groups were compared in terms of operation time, intraoperative bleeding, and Rasmussen scores for knee function and medial angle of the proximal tibia at the last follow-up. Results: The 2 groups were comparable due to insignificant differences in the preoperative general data between them (P>0.05). The tractor and manipulation groups were followed up for a mean of 9.8 months and 10.9 months respectively. Bone healing was achieved in all patients. For the tractor and manipulation groups, respectively, the operation time was127.8±40.8 min and 174.1±66.8 min, and the intraoperative bleeding 115.6±41.7 mL and 184.3±91.4 mL, showing significant differences between them (P<0.05). In the tractor group at the last follow-up, the knee flexion was 126.1°±9.3°, the medial angle of the proximal tibial 87.8°±1.4°, the posterior inclination angle 8.7°±3.8° and the Rasmussen score 26.5±2.9 points; in the manipulation group at the final follow-up, the knee flexion was 124.7°±8.9°, the medial angle of the proximal tibial 86.9°±2.1°, the posterior inclination angle 9.8°±4.1° and the Rasmussen score 25.6±3.2 points. There were no significant differences between the 2 groups in the above items (P>0.05). Loss of force line was observed in none of the patients at the last follow-up. Conclusion Compared with conventional manipulation reduction, use of our self-designed lower limb axial tractor can shorten operation time, reduce bleeding and facilitate reduction of fracture fragments in the treatment of tibial plateau fractures of Schatzker types Ⅴ and Ⅵ.

Objective To analyze the complications of transvaginal natural orifice transluminal endoscopic surgery( TV-NOTES) nephrectomy, and to explore effective measures to prevent and manage those complication.Methods From May 2010 to January 2015, a total of 178 females who had been married and given birth underwent TV-NOTES nephrectomy in our center.The average age was 47 ( ranging 23 to 71 ) years and the average BMI was 23.6 ( ranging 14.7 to 31.9 ) kg/m2.Pathological diagnosis included 142 cases of non-functional kidneys, 29 cases of renal tumors ( T1 N0-1 M0 25 cases, T2 N0-1 M0 4 cases) , and 7 cases of renal tuberculosis.One hundred and sixty hybrid TV-NOTES nephrectomy procedures (simple in 132, radical in 28) and 18 pure TV-NOTES nephrectomy procedures (simple in 17, radical in 1) were performed.Intraoperative and postoperative complications were graded according to Satava and Clavien-Dindo grade classifications.The major complications and relative treatments were analyzed.Results Among the 178 TV-NOTES nephrectomy procedures, there were 40 ( 22.5%) complications occurred, including 13 (7.3%) major complications, in which there were 11 cases of intraoperative complications (6.2%),2 cases of postoperative complications (1.1%).All the complications were successfully managed using organ repair or resection, embolectomy, hemostasis, and so on.No intraoperative and postoperative deaths occurred.There was no significant difference in major complications between hybrid TV-NOTES and pure TV-NOTES nephrectomy (6.9%vs.11.1%, P=0.620).The intraoperative major complications in the early developmental stage of TV-NOTES nephrectomy were more than that of the late stage (20.0%vs.3.4%, P=0.004).There was no significant difference in postoperative complications between the early and late developmental stage of TV-NOTES nephrectomy (13.3%vs.10.8%, P=0.751).Conclusions TV-NOTES nephrectomy is safe and feasible, but there are some major complications worthy of attention.Personal prevent and treatment strategy should be considered.

OBJECTIVETo evaluate the effects of polymorphisms in XRCC1 and APE1 genes on vinyl chloride (VC)-induced chromosomal damage in peripheral lymphocytes.

METHODSIn this study, 317 workers occupationally exposed to VC were recruited from a factory in Shandong Province, China. The micronucleus (MN) frequency in peripheral lymphocytes was used as an indicator of chromosomal damage. Polymerase chain reaction-restriction fragment length polymorphism and created restriction site combined with restriction fragment length polymorphism were used to determine the five single nucleotide polymorphisms in XRCC1 and APE1 genes in the base excision repair pathway. The association of chromosomal damage with these polymorphisms and the haplotype of XRCC1 was analyzed using Poisson regression and PHASE 2.0.2.

RESULTSIt was found that among the VC-exposed workers, individuals with XRCC1 polymorphisms (-77C/T, Arg194Trp, Arg280His, and Arg399Gln) had a significantly higher MN frequency than those with homozygous wild-type genotypes, with frequency ratios (FR) as follows, respectively: FR = 1.21, 95%CI: 1.05∼1.39 (P < 0.05); FR = 1.14, 95%CI: 1.00∼1.38 (P < 0.05); FR = 1.26, 95%CI: 1.11∼1.44 (P < 0.05); FR = 1.23, 95%CI: 1.08∼1.46 (P < 0.05). APE1 Asp148Glu was found of no significant relationship with MN frequency. Haplotype analysis of XRCC1 demonstrated that the MN frequencies in subjects with CTAA/CTAA and CCAA/CTAA were significantly higher than that in those with TCGG/TCGG (FR = 1.19, 95%CI: 1.02∼1.32, P < 0.05; FR = 1.41, 95%CI: 1.02∼1.87, P < 0.05). Furthermore, association was found between accumulated exposure to VC and XRCC1 polymorphisms (-77C/T, Arg194Trp, Arg280His, and Arg399Gln) after adjustment for age, sex, drinking, and smoking.

CONCLUSIONVC can induce chromosomal damage even when the exposure level is lower than the national occupational health standard of China (PC-TWA: 10 mg/m(3)); the polymorphisms in XRCC1 and APE1 are associated with chromosomal damage induced by VC.

Adult ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; DNA-Binding Proteins ; genetics ; Female ; Haplotypes ; Humans ; Male ; Micronuclei, Chromosome-Defective ; Middle Aged ; Occupational Exposure ; adverse effects ; Polymorphism, Restriction Fragment Length ; Vinyl Chloride ; poisoning ; X-ray Repair Cross Complementing Protein 1 ; Young Adult

Objective:To explore the clinical application and triage management value of using blood circulating cell-free DNA (cfDNA) (cysteine dioxygenase type 1 gene, CDO1, and Homeobox protein A9 gene, HOXA9) hypermethylation level to detect and diagnose ovarian cancer.Methods:A case-control study was conducted on patients who went for surgery at Chengdu Womens and Childrens Central Hospital from November 2022 to October 2023. Blood samples were collected before surgery for evaluation of cancer antigen 125 (CA125), human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA) score, and DNA methylation testing. The basic clinical information, biomarkers, and transvaginal ultrasound (TVS) information were collected simultaneously. Information from a total of 151 patients was collected, including 122 cases with benign pathology and 29 ovarian cancer cases. The pathologic diagnosis of ovarian tissue was defined as the gold standard. The multivariate logistic regression analysis was used to identify high-risk factors for ovarian cancer. The clinical efficacy of DNA methylation detection for ovarian cancer was analyzed using the area under curve (AUC).Results:The results showed that the age, menopausal status, CA125 and HE4 detection, ROMA score, positivity rate of CDO1 gene and HOXA9 gene single or combined testing in ovarian cancer patients were higher than those in the benign group and showed significant differences ( P<0.05). Among these detection protocols, the AUC of CDO1 and HOXA9 dual gene methylation testing for ovarian cancer was the highest at 0.936 (95% CI, 0.878-0.994), with 89.7% (95% CI 73.6%-96.4%) sensitivity and 97.5% (95% CI 93.0%-99.2%) specificity, respectively. The positive detection rate of CDO1 and HOXA9 dual gene methylation in early ovarian cancer FOGO I-II stage is 12/14 higher than other tests. Conclusion:Blood cfDNA methylation detection, a simple, non-invasive, and highly sensitive detection method, is superior to the current ovarian cancer testing in the risk assessment and early detection.

Objective To investigate the effect of periosteum deficiency on the treatment of segmental bone defect of lower extremity by Ilizarov bone transport Technique.Methods From April 2009 to April 2016,data of 54 cases with lower limb segmental bone defect who were treated by Ilizarov bone transport technique were retrospectively analyzed.According to intraoperative observation of the distribution of periosteum at the osteotomy site,all cases were divided into two groups.There were 10 cases in absent periosteum group,including 6 males and 4 females,aged from 23 to 57 years old (mean,38.20±9.90 years old).There were 3 cases locating at femur,and 7 cases at tibia.4 cases had non-infective bone defect,while the other 6 had infective bone defect.Segmental bone defect ranged from 4.0 to 14.0 cm (mean,6.85±2.87 cm).There were 44 cases in normal periosteum group,including 32 males and 12 females,aged from 22 to 65 years old (mean,38.90±10.10 years old).There were 9 cases locating at femur and 35 cases at tibia.12 cases had non-infective bone defect,while 32 cases had infective bone defect.Segmental bone defect ranged from 4.0 to 9.0 cm (mean,6.09±1.54 cm).Visual analogue scale (VAS) was used to estimate the pain during bone transport.The distraction index,bone healing index,external fixator index was collected to evaluate the distraction osteogenesis and mineralization.The Paley method was utilized to appraisal the function of proximal joints in bone transport.Results All the 54 cases were followed up for 24-48 months,with an average of 28.50±5.70 months.All skin and soft tissue defects were satisfied healed.All limbs were restored to their original lengths,and bone healing was eventually achieved.There were no statistically significant differences in the distraction index,bone healing index,external fixator index,VAS between absent periosteum group (12.29± 1.04 d/cm,39.00±3.96 d/cm,51.25±3.69 d/cm,3.30± 1.77 score) and normal periosteum group (11.67±0.96 d/cm,38.07±4.22 d/cm,49.74±4.25 d/cm,3.36± 1.66 score)(P ＞ 0.05).Paley adjacent joint function evaluation showed no statistically difference between absent periosteum group [good rate 100％(10/10)] and normal periosteum group [good rate 97.7％ (43/44)] (P=0.901).The overall incidence of complications was 20.0％ (2/10) in the absence of periosteum group,with 1 case of mild infection and 1 cases of tissue contracture incarcerated at the docking site.The overall incidence of complications was 27.3％ (12/44) in the normal of periosteum group,with 2 cases of mild infection,3 cases of poor limb strength alignment,6 cases of tissue contracture incarcerated at the docking site and 1 case of severe foot drop deformity.There was no statistically significant difference in the overall incidence of complications between the two groups (P=0.636).Conclusion The absence of periosteum at the epiphyseal osteotomy site hasno significant effect on Ilizarov bone transport technique in the treatment of segmental bone defects of lower limbs in bone regeneration and healing,mineralization and bone remodeling,and does not increase the incidence of complications.