Objective: To explore the alternatives for T-tube placement in surgical treatment of bile duct stones.Methods: Fourty-nine patients with bile duct stones have been performed retrograde transhepatic biliary drainages(RTBD)since 1991.Among them,37 cases were placed the drainage tubes through the right hepatic lobes by retrograde punctures;the other 12 cases with intrahepatic stones were placed the drainage tubes through the intrahepatic bile duct stumps following partial hepatectomies.All the CBD exploration incisions were primarily closed.Results: (1)RTBD by retrograde puncture:Two postoperative complications(5.4%,pneumothorax and biliary hemorrhage)occured.Both cases were cured by medical therapy.The drainage tubes were removed on the 8th postoperative day.(2)RTBD through intrahepatic ducts:Four cases had residual stones(30%)which had been cleared up by combination of extraction and intraluminal lithotripsy.Nineteen cases(38.8%)had the gallbladders preserved.Postoperative cholangiographies showed that all the gallbladders were smooth and well-filled and the extrahepatic bile ducts were in normal shapes.Extrahepatic bile ducts and all the preserved gallbladders did not show any deformity and stone formation by ultrasonography during 6 months to 9 years'following-up.Conclusion: RTBD by retrograde puncture has the advantage of shorter period of bearing drainage tubes;RTBD through intrahepatic duct stump after partial hepatectomy makes it easier to extract postoperative residual stones.Both methods have no negative impacts on the anatomies of both the extrahepatic bile duct and the gallbladder.

Background and Objectives: This study was to investigate the role of microRNA-29a-3p (miR-29a-3p) in human bone marrow mesenchymal stem cells (hBMSCs), and its relationship with steroid-associated osteonecrosis. Methods: and Results: The online tool GEO2R was used to screen out the differentially expressed genes (DEGs) in GSE123568 dataset. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-29a-3p, forkhead box O3 (FOXO3), alkaline phosphatase (ALP), bone gamma-carboxyglutamate protein (OCN) and RUNX family transcription factor 2 (Runx2) in the hBMSCs isolated from the patients with steroid-associated osteonecrosis. CCK-8 assay was executed to measure cell viability; western blot assay was utilized to detect FOXO3, ALP, Runx2, OCN and β-catenin expression. Cell apoptosis and cell cycle were detected by flow cytometry. Immunofluorescence assay was used to detect the sub-cellular localization of β-catenin. Bioinformatics analysis and luciferase reporter gene assay were performed to confirm whether miR-29a-3p can combine with FOXO3 3’UTR. MiR-29a-3p was markedly up-regulated in the hBMSCs of patients with steroid-associated osteonecrosis, while FOXO3 mRNA was significantly down-regulated. Transfection of miR-29a-3p mimics significantly inhibited the hBMSCs’ proliferation, osteogenic differentiation markers’ expressions, including ALP, Runx2, OCN, and repressed the ALP activity, as well as promoted cell apoptosis and cell-cycle arrest. FOXO3 was identified as a target gene of miR-29a-3p, and miR-29a-3p can inhibit the expression of FOXO3 and β-catenin, and inhibition of miR-29a-3p promoted translocation of β-catenin to the nucleus. Conclusions MiR-29a-3p can modulate FOXO3 expression and Wnt/β-catenin signaling to inhibit viability and osteogenic differentiation of hBMSCs, thereby promoting the development of steroid-associated osteonecrosis.

To obtain active protein of pIL-18 expression in Lactococcus lactis, and to observe its biological activity, the total RNA was extracted as template from peripheral blood mononuclear cells. Porcine interleukin 18 (pIL-18) was amplified by RT-PCR. The resulting fragment was cloned into pAMJ399 L. lactis vector, and then transformed to L. lactis MG1363 cells by electroporation. Expression of pIL-18 protein was detected by SDS-PAGE and Western-blotting. Bioactivity of the product was tested by pig spleen lymphocyte proliferation test and cytopathogenic effect inhibition assay. The result of Western blotting and bioactivity test shows that the molecular weight of pIL-18 protein was 19 kDa. The react line was observed in both supernatant and precipitated of the recombinant bacteria pAMJ399-pIL18/MG1363. The expressed pIL-18 can promote the proliferation of pig spleen lymphocyte, and significantly inhibit virus multiplication. As conclusion, porcine interleukin-18 was successfully expressed in L. lactis, and the product was biologically active.
Animals ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Electroporation ; Genetic Vectors ; Interleukin-18 ; biosynthesis ; Lactococcus lactis ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

Currently liver transplantation is the most effective treatment options for patients with end-stage liver disease, but the lack of donor livers limits the development of liver transplantation. With the increasing usage of marginal donor liver such as fatty donor liver, therefore how to make marginal donor liver play a better role has become a new research hotspot. Although the use of fatty liver as a donor can effectively alleviate the contradiction between recipient demand and donor organ shortage, the risk of early post-transplant graft dysfunction also increases. As donor of fatty liver, the degree of steatoidosis should be moderate or below as far as possible, which significantly reduces postoperative complications and promote the recovery of recipients. Meanwhile, it is essential to monitor the lipid indicators post the surgery, and drug intervention can be added if necessary, which may improve the function of the graft.

Objective:To explore the application of failure mode and effects analysis (FMEA) in low-energy X-ray intraoperative radiotherapy (IORT), analyze its potential risks in IORT, and preliminarily explore the feasibility of FMEA in optimizing IORT management and reducing the occurrence of potential risks.Methods:An FMEA working group was established by the IORT team (1 radiologist, 1 radiology physicist, 2 surgeons, and 2 nurses) to apply the FMEA methodology to conduct a systematic risk assessment. The process modules were established, the potential failure modes and causes for each module were analyzed, the severity (SR), frequency of occurrence (OR) and likelihood of detection (DR) of failure modes were scored and the risk priority number (RPN) was calculated: RPN= SR × OR × DR. The possible errors and potential clinical impact of each part of the radiotherapy process were prospectively analyzed and understood, the causes and current measures were analyzed for each failure mode and preventive measures were proposed and risk management measures were taken accordingly.Results:The IORT process was divided into 8 modules with 14 failure modes. The highest OR value was unsatisfactory target area confirmation (7 points), the highest SR value was equipment failure to discharge the beam (10 points), the highest DR value was wrong key entry after dose calculation (7 points), the highest RPN values were unsatisfactory target area confirmation (210 points) and ineffective protection of endangered organs (180 points). Weaknesses were corrected according to priorities, workflows were optimized and more effective management methods were developed.Conclusion:FMEA is an effective method of IORT management and contributes to reducing the occurrence of potential risks.

Objective:To provide a theoretical basis for diagnosing and treating recurrent Kawasaki disease based on data analysis of clinical symptoms.Methods:Data analysis of children with recurrent Kawasaki disease admitted to Beijing Children′s Hospital of Capital Medical University from January 2016 to December 2019 was conducted, including comparisons of the initial onset and the recurrence on patients′ clinical features, auxiliary examination and treatment.Results:During the four-year scale, 3 041 children with Kawasaki disease were admitted to the department of Cardiology.The recurrence data involved 65 children[male∶female 3.1∶1, average aged(2.42 ± 2.04) years]. First, children′s fever duration was(5.66 ± 2.37) days in recurrence data, significantly shorter than that of their initial onset.The recurrence data also reported a lower incidence of rash and limb changes, together with respiratory and gastrointestinal symptoms.Then, 19 cases out of the 65 children got coronary artery lesion(CAL) at their initial onset.A relief of CAL was reported from 15 children when the disease recurred, along with 11 new-reported CAL cases.Intravenous immunoglobulin(IVIG) refractory Kawasaki disease cases accounted for nine at the initial onset and 12 at the recurrent onset, respectively.Five IVIG refractory recurrent cases reported significant relief after 2 g/kg IVIG treatment.Conclusion:The recurrent Kawasaki disease in children usually occurs among children under three years old, within the first year after the initial onset.Most of the recurrent cases report a shorter fever duration and less typical clinical symptoms than their first onset.In some cases, IVIG treatment showed effects initially but failed to work when the disease recurred.Therefore, glucocorticoid or infliximab should be considered for further treatment.

Objective To develop an intelligent management system of documents and records that meets the requirements of ISO 15189 quality management system applied to regional medical laboratory center.Methods Based on the relevant terms of ISO 15189 document and record,an intelligent management system for laboratory documents and records with B/S architecture was established by the help of computer technology.Results The document and record management system stored the quality system documents required for ISO 15189 review as well as the external documents,such as laboratory-related laws and regulations,standard guidelines and expert consensus,which can be accessed by users through a computer or mobile phone.The multi-process control mode was adopted to author-ize the special personnel to complete the compiling,review,approval and,release of the files according to the type and attribute of the documents.The management system of records established 9 special records,319 form records and 20 process records to support the ef-fective operation of the quality system.Conclusion The established intelligent management system for documents and records has fully met the requirements of ISO 15189 on documents and records management,realized the sharing and interconnection of the documents in the given region.It is convenient for the employees to consult the documents and fill in the records in a timely and efficient manner.

Animals ; B7-H1 Antigen ; immunology ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; immunology ; Humans ; Interferons ; immunology ; Mice ; Neoplasm Proteins ; immunology ; Neoplasms ; immunology ; Up-Regulation ; immunology

Objective:To analyze locoregional recurrence (LRR) pattern of patients with pT 1-2N 1 breast cancer after modified radical mastectomy, with and without adjuvant radiotherapy (RT). Methods:A total of 5442 eligible patients with breast cancer from 12 Chinese centers were included. The LRR sites and the effect of RT at different sites on recurrence in patients with and without RT were analyzed. The Kaplan-Meier method was used to calculate the cumulative LRR rate, and the difference was compared by the log-rank test.Results:With a median follow-up time of 63.8 months for the entire cohort, 395 patients developed LRR. The chest wall and supraclavicular fossa were the most common LRR sites, regardless of RT or molecular subtypes. The 5-year chest wall recurrence rates for patients with and without chest wall irradiation were 2.5% and 3.8%( P=0.003); the 5-year supraclavicular lymph nodal recurrence rates for patients with and without supraclavicular fossa irradiation were 1.3% and 4.1%( P<0.001); the 5-year axillary recurrence-free rates for patients with and without axillary irradiation were 0.8% and 1.5%( HR=0.31, 95% CI: 0.04-2.23, P=0.219); and the 5-year internal mammary nodal recurrence-free rates for patients with and without internal mammary nodal irradiation were 0.8% and 1.5%( HR=0.45, 95% CI: 0.11-1.90, P=0.268). Conclusions:The chest wall and supraclavicular fossa are the most common LRR sites of patients with pT 1-2N 1 breast cancer after modified radical mastectomy, which is not affected by adjuvant RT or molecular subtypes. The chest wall and supraclavicular fossa irradiation significantly reduce the risk of recurrence in the corresponding area. However, axillary and internal mammary nodal irradiation has no impact on the risk of recurrence in the corresponding area.