Objective To construct a shuttle plasmid for inducible gene expression in Borrelia burgdorferi (B.burgdorferi) with an advantage of flexible genetic manipulation.Methods The IPTG-inducible lac repressor/operator system from Escherichia coli (E.coli) was adopted and modified in the current study.The plasmid shuttle vector was developed by inserting multiple cloning sites,FLAG and HA tags into the shuttle vector by molecular cloning approaches.The target gene was inserted at the site under the control of the promoter (Tn5 derivate) in plasmid pQE30.This promoter contained two lac operators and a codonoptimized lacI gene driven by flaB promoter.Results A plasmid shuttle vector,pJJ275,was successfully constructed with the ability to express target genes in B.burgdorferi in the presence of IPTG.By using this system,a HA-tagged rpoS gene was introduced into the typical infectious strain B.burgdorferi B31.The target gene expression induced by IPTG was confirmed at transcriptional and translational levels.The RpoS dependent virulence factor of Borrelia,OspC,was also detected,indicating that the expressed protein was functional.Conclusion The constructed plasmid shuttle vector can express exogenous genes in B.burgdorferi with an inducible feature and an advantage of flexible genetic manipulation.It can be applied for genetic manipulation of B.burgdorferi involved in gene regulation and complementation.

Objective To investigate the clinical features and treatments of pituitary abscess.Method The clinical data of 6 patients with pituitary abscess were examed along with a review of the literature.Results Of 6 patients,headache was presented in 5 patients,hypopituitarism in 4 patients,visual disturbance and/or bitemporal hemianopsia in 4 patients and fever in 1 patient.MRI and CT images showed a cystic sellar lesion with ring enhancement in 5 patients.Preoperative diagnosis of pituitary abscess was made in 2 patients,pituitary adenomas in 3 patients and craniopharyngiomas in 1 patient.All cases were treated surgically by transsphenoidal approach in 5 patients and transscranial in 1 patient.Followed with postoperative antibiotics therapy for 3 weeks,the symptoms were improved postoperatively in all cases.Followed up 8 months to 10 years,1 patient who underwent craniotomy recurred and wag cured by via transsphenoidal surgery.Conclusions The pituitary abscess is easily misdiagnosed.The cystic pituitary lesion should be considered the possibility of pituitary abscess.Transsphenoidal surgery and proper perioperative antibiotics therapy are the keys to the treatment of pituitary abscess.

Objective To investigate and evaluate the clinical value of full endoscopic transnasal transphenoidal approach for the surgery of pituitary adenomas.Methods Seventy-two patients,who underwent full endoscopic transnasal transphenoidal approach for the surgery of pituitary adenomas,were selected from the Medical Center of Pituitary Adenomas of our hospital from January 2009 to March 2012.To retrospectively investigate pre- and post-operation symptoms,hormone levels,images information,operation information,complications,following-up information and so on.Results Among the 72 consecutive patients,there were 22 nonfunctioning adenomas,twenty-four prolactin secreting adenomas,seven somatotropin secreting adenomas,five adrenocorticotropic hormone secreting adenomas,one thyrotropin secreting adenomas,and 13 multi-secreting adenomas.The tumor removal was total in 56(77.8％),subtotal 13(18.0％),and partial 3(4.2％).Five cases had CSF leaks,and 6 diabetes insipidus.After 3-24 months of follow-up,the levels of increasing-hormone declined to normal levels in most patients.Conclusion Full endoscopic transnasal transphenoidal approach for the surgery of pituitary adenomas is a kind of technique which is safe,minimally invasive,having less complications and fast recovery.However,it is necessary for surgeons to accept systematic and specialized training,and own advanced equipments.

Objective To investigate the significance of computed tomography(CT)and 3.0 T magnetic resonance imaging(MRI)in intensity-modulated radiotherapy(IMRT)for esophageal carcinoma. Methods Thirty-five patients newly diagnosed with esophageal carcinoma who received radical radiotherapy in our hospital from November 2013 to April 2015 were enrolled as subjects. Target volume was delineated on the CT images and MRI images(T2-weighted and diffusion-weighted fusion images). The MRI-and CT-based IMRT plans were designed using the same dose prescription and dose constraints for organs at risk(OAR). The target volume,prescribed dose,and doses for OAR were compared between the two plans. Results In the two plans, dose distribution and planning parameters met the clinical requirement. The length of lesion,gross tumor volume (GTV),and planning target volume(PTV)defined by 3.0 T MRI were significantly smaller than those defined by CT(P=0.00,0.03,0.03). There were no significant differences in the D 2%,D 98%,D 50%,homogeneity index,or conformity index for primary GTV(PGTV)and PTV-PGTV between the two plans(all P>0.05). Compared with the CT-based plan,the 3.0 T MRI-based plan had a significantly smaller mean dose to the lungs and an insignificantly smaller actual dose to the lungs(P=0.00;P>0.05).There were no significant differences in maximum doses tolerated by the spinal cord or heart between the two plans. Conclusions In terms of target volume delineation and dosimetric parameters, both CT-and 3.0 T MRI-based plans meet the clinical requirement. The 3.0 T MRI-based plan may provide potential benefits for some OAR due to a smaller target volume compared with the CT-based plan.

Objective To explore the early clinical efficacy of 3D printing technology-assisted customized prosthesis with preservation of femoral condyles for the reconstruction of bone defects after resection of distal femoral malignancies.Methods From June 2019 to September 2021,our department determined the lesion range of 12 cases of malignant tumors in the distal femur through CT and MRI before surgery.We used computer-aided production of osteotomy guide plates and customized prostheses that preserved the femoral condyle.After the prostheses were customized,the tumor segment was removed and a reconstruction was performed by using the customized prosthesis with preservation of femoral condyles.The knee function was evaluated by the American Musculoskeletal Tumor Society(MSTS)functional evaluation criteria.Results The operative time was 180-270 min,with a mean of 200 min.The intraoperative bleeding was 300-800 ml,with a mean of 450 ml.A total of 12 cases were followed up for 12-50 months(average,21.9 months).Two patients died of lung metastasis at 12 and 16 months after surgery,one patient survived with lung metastasis at 3 months after surgery,and one patient(8.3%)developed periprosthetic infection at 1 year after surgery.In 10 patients,imaging showed that the prosthesis and plate were in a good position without signs of loosening or bone resorption,the proximal femur was fixed in place with the 3D printed collar,and continuous bone scab formation was found in 4 cases.At the final follow-up,the functional status of the affected limb was good,with the knee flexion of 105°-135°(mean,115°)and no limitation of extension.The MSTS score was 22-28 points[mean,(24.3±1.8)points].Conclusion Application of 3D printing-assisted customized prosthetic replacement technology with preservation of the femoral condyles allows for precise resection of malignant distal femoral tumors and effective anatomical reconstruction without increasing surgical trauma and risk,significantly improving postoperative limb function and potentially reducing long-term complications.

Objective To investigate the effect of nutritional intervention upon the clinical efficacy of chemoradiotherapy in patients diagnosed with esophageal carcinoma. Methods A total of 46 patients who were diagnosed with esophageal cancer in Anhui Cancer Hospital from November 2016 to August 2017 were enrolled in this prospective study. All patients were randomly and evenly divided into the nutritional intervention (NI) and routine treatment (RT) groups. The changes in body mass index (BMI),PG-SGA, serum albumin ( ALB), hemoglobin ( HB), white blood cell ( WBC) and other objective nutritional parameters and the incidence of chemoradiotherapy-induced complications were recorded before and after chemoradiotherapy. Results Prior to chemoradiotherapy,age,sex,BMI,ALB,PLT and clinical staging did not significantly differ between two groups (all P>0. 05).In the NI group,the BMI was (21.52±2. 67) after chemoradiotherapy,significantly higher than (21.13±2. 73) before radiotherapy (P= 0. 000).Moreover,the PG-SGA score after chemoradiotherapy was significantly lower compared with that before chemoradiotherapy (P= 0. 000).In the RT group,the BMI,Hb,ALB,PLT and WBC after chemoradiotherapy were significantly lower than those before radiotherapy, and thePG-SGA score was worse after chemoradiotherapy ( all P<0. 05).In the NI group, the incidence of grade 3 myelosuppression was 4. 34％, significantly lower than 8. 68％ in the RT group ( P= 0. 000 ). Conclusions Patients with esophageal cancer treated with chemoradiotherapy have a high nutritional risk. Nutritional intervention can improve the nutritional status, reduce the incidence of chemoradiotherapy-induced complications,and probably improve the quality of life and clinical prognosis.

Objective:This study evaluates the performance of chemiluminescence assay, which is designed to detect Hepatitis D Virus (HDV) Immunoglobulin G (IgG) antibodies.Methods:A comparative analysis was conducted among chemiluminescence anti-HDV IgG reagent, the magnetic particle-based domestic reagent A and domestic reagent B, and the Robo Gene HDV RNA kit, using 1909 HBsAg-positive plasma samples. This comparison aimed to delineate clinical specificity and detection accuracy. The anti-HDV IgG reagent precision was assessed at three different concentration levels following the Clinical Laboratory Standards Institute EP5-A2 guidelines. The specificity of the assay was validated using 200 HAV IgM positive, 545 HBsAg-positive but anti-HDV IgG-negative, 350 anti HCV positive plasma samples and 200 healthy human blood samples. Additionally, a concordance study was conducted with 545 HBsAg-positive and 37 anti-HDV IgG-positive plasma samples, comparing the anti-HDV IgG reagent against reagent A.Results:1 909 HBsAg-positive plasma samples were tested using 3 anti HDV IgG reagent and 1 HDV RNA reagent, 19 samples were identified as anti-HDV IgG-positive. The anti-HDV IgG demonstrated superior accuracy and specificity. The assay exhibited excellent precision, with intra-assay coefficient of variation (CV) values ranging from 1.57% to 4.30%, and inter-assay CV values between 1.71% and 4.67% for detecting samples at high, medium, and low concentration levels. Concordance with Reagent A showed consistent results in both positive and negative detections.Conclusion:In this study, the anti-HDV IgG reagent (chemiluminescence method) displayed outstanding specificity in detecting clinical samples and exhibited a high conformity rate with commercialized reagents, making it potentially suitable for screening anti-HDV IgG in HBsAg-positive samples.

Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.