Proteomic analysis of core needle biopsy (CNB) sample from patient populations is critical to our understanding of human disease,but has been hindered by its particular small size.Here,we present a method for the proteomic analysis of CNB sample based on the two dimensional electrophoresis.Proteins were extracted directly from 3 rat liver CNB specimens and a human prostate CNB sample.respectively.24 cm Immobiline DryStrip (pH 3-10NL) and 12.5% SDS-PAGE were introduced to separate the proteins.Interesting spots were analyzed by MALDI TOF/TOF mass spectrometry after tryptic digestion.With this method,consistent electrophoretic patterns of more than 2 500 protein spots were reproducibly obtained after silver staining,from rat liver CNB specimens.Qualitatively and quantitatively reproducible results also yield when the method was applied to a human prostate CNB sample.57 stochastically selected protein spots were analyzed by MALDI TOF/TOF moss spectrometry.and were identified with high confidence including faint ones.This simple and reproducible approach raises the opportunity of defining key molecular events of human disease pathologies.

OBJECTIVETo investigate the rationality and feasibility of primary closure of the common bile duct after choledochotomy for common bile duct calculi.

METHODSFrom January 1990 to June 2001, 386 patients with the evidence of stones in the common bile duct underwent choledochotomy. Among them, 215 received primary closure of the common bile duct (group A) and 171 T-tube drainage (group B). The patients with emergency operations were excluded. Intraoperative choledochoscopy or cholangiography was routinely performed to rule out the possibility of retained stones. The duct was meticulously stitched using 0/3 to 0/5 absorbent sutures for primary closure. A T-tube was placed in the subhepatic space in the patients of both groups.

RESULTSPostoperative bile leakage was seen in 9 patients of group A and in 5 of group B, respectively (P > 0.05), and no reoperations were necessary. After surgery, the average time and volume of transfusion was 4.9 days and 9.1 liters in group A, versus 7.3 days and 12.8 liters in group B (P < 0.01). The patients in group B had a longer postoperative hospital stay than the those in group A (average 17.6:10.0 days, P < 0.01). T-tube removal resulted in bile peritonitis in 5 patients at day 16, 17, 19, 21 and 22 after surgery in group B, and 3 patients required repeated surgery.

CONCLUSIONSPrimary closure of the common bile duct after choledochotomy is safe, effective, and inexpensive in selected patients with common bile duct calculi, and should be regarded as an alternative procedure.

Adult ; Aged ; Biliary Tract Surgical Procedures ; methods ; Choledocholithiasis ; surgery ; Common Bile Duct ; surgery ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Objective:To investigate the application value of metagenomic next generation sequencing (mNGS) in the diagnosis of spinal infectious diseases except tuberculosis and non-tuberculous mycobacterium infection.Methods:According to the inclusion and exclusion criteria, the clinical data of 171 patients with suspected non-tuberculous infectious diseases of spine from January 2019 to November 2020 were analyzed retrospectively. All patients underwent puncture biopsy of spine or surgery to obtain tissue specimens, then tissue biopsy were measured, and Foci purulent blood or lavage fluid was obtained for routine bacterial and mNGS. The differences of reporting time, sensitivity (positive rate) and specificity (true negative rate) were compared between the two methods. The pathogenic microorganism spectrum of spinal infection was analyzed. The effects of specimen collection method, preoperative antibiotic use and specimen type on the detection of pathogenic microorganisms were analyzed.Results:According to the diagnostic criteria of non-tuberculous infectious diseases of the spine, there were 136 cases of infection, among which 111 cases had specific pathogenic microorganism. The average reporting time of traditional culture and mNGS were 81.67+15.52 h and 36.33+11.92 h. There were 43 positive cases and 5 false positive cases in traditional culture, the sensitivity was 31.62%, and the specificity was 85.71%; There were 76 positive cases and 19 false positive cases in mNGS, the sensitivity was 55.88%, and the specificity was 45.71%. There were significant statistical differences in the sensitivity, specificity and reporting time for detection of pathogenic microorganisms by mNGS. The top 3 pathogens of non-specific infection were Staphylococcus aureus and Staphylococcus, Escherichia coli and Streptococcus. Eleven patients with non-tuberculous specific infection were confirmed, including 2 positive cases with traditional culture, 11 positive cases with mNGS. A higher detection rate for rare specific infections were expressed in mNGS. Logistic regression analysis indicated that the standard antibiotic use 4 weeks before surgery and specimen acquisition methods had significant effects on the traditional culture results, while the specimen acquisition methods had no statistically significant effects on the mNGS.Conclusion:Metagenomic next generation sequencing has a higher sensitivity to the detection of pathogenic microorganisms in the diagnosis of non-tuberculous spinal infectious diseases, especially for the detection of rare pathogenic microorganisms, which has a high diagnostic value.