Child ; Gene Rearrangement, T-Lymphocyte ; Humans ; Neoplasm, Residual ; diagnosis ; Polymerase Chain Reaction ; methods ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; Receptors, Antigen, T-Cell ; genetics

Adolescent ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma

SUMMARY To investigate the effect and feasibility of total laparoscopy to treat hepatolithiasis using gall-bladder-hepatic duct subcutaneous tunnel.Retrospective analysis was conducted of the case data of 11 pa-tients with hepatolithiasis who underwent total laparoscopic treatment using gallbladder-hepatic duct sub-cutaneous tunnel from January 2010 to October 2014.The operation time,blood loss,postoperative com-plications and recurrence of stones were recorded.All the cases completed the operation.The average hos-pital-stay was 9.2 days (range:3 -29 d).The average operation time was 298 min (range:225 -480 min).The average blood loss was 253 mL (range:50 -700 mL),and the average blood loss of liver re-section groups was 325 mL (range:200 -700 mL).The average discharge time was 3.3 days (range:3 -5 d).The rate of postoperative residual stones was 36.4% (4 /11).We extracted stones with chole-dochofiberscope via T-tube sinus six weeks after operation.One case developed biliary leakage,and healed through adequate drainage and the T-tube was pulled out after one month.There was no periopera-tive mortality.All the cases were followed up and the mean follow-up was 22 months (range:2 -51 months).The anastomotic stenosis of gallbladder-hepatic duct was found in one case.But we got a good therapeutic result with performed gallbladder chemical ablation with 95% ethanol.No recurrence of hepa-tolithiasis was found.As a choice for minimally invasive method to hepatolithiasis using gallbladder-he-patic duct subcutaneous tunnel,total laparoscopy is a safe and feasible procedure.

Objective To lessen the learning curve of percutaneous renal lithotripsy(PCNL),we introduced a novel ex-vivo learning and training model for PCNL under fluoroscopy and ultrasonography-guided access. Methods The model was set up nailing an adult porcine kidney with＞3 cm ureter (freshly removed from the slaughtering factory),with a full thickness flap of the thoracic wall or abdomen wall with two ribs,to a board.The porcine kidney was placed within the flap with the catheterized ureter outside.The kidney was enclosed by the flap so as to create a roodel for percutaneous renal surgery;with the ribs overlying the upper portion of the kidney.The model was fixed to the board by two nails.Artificial stone material was implanted in the renal pelvis.Fluoroscopy guidance access:Retrograde pyelography via injection of contrast medium into the ureteral catheter images the collecting system. After the long axis of the target calyx is identified,the C-arm is rotated 30 degrees toward the surgeon,placing the C-arm axis in the same posterior plane of the kidney.The needle is advanced in the plane of the fluoroscopic beam,and the appropriate needle placement is determined by obtaining a bull's-eye sign on the fluoroscope screen. Rotating the C-arm to a vertical position monitors the depth of the needle penetration. Ultrasonography guidance access:The renal pelvis can he filled with normal saline through a catheter to simulate hydronephrosis and the target calyx is identified under ultrasonography guidance.The tract dilation and stone disintegration were followed.After training,the kidney can be opened to examine the target calyx and the complication of dilation. Results Altogether,126 urologists attended a urologic endoscopic technique training course.Of the 126 trainees,104 (82.5%) successfully performed the whole percutaneous procedure.At the end of training,114 (90.5 %) of the 126 attendees rated the porcine kidney model for simulation of percutaneous renal surgery as very helpful or helpful.Conclusions This biological training model simulates realistically the clinical procedure of PCNL with respect to trainee experience in a low stress environment that provides an opportunity for repetitive performances in order to learn basic technical skills for the clinical procedure of PCNL in the future.

OBJECTIVETo evaluate the efficacy and safety of the combination therapy of Xipayimaizipizi Capsules and Tamsulo- sin in the treatment of benign prostatic hyperplasia (BPH).

METHODSWe randomly assigned 60 BPH patients to a control and a combination group of equal number, the former aged 62.03 ± 10.19 years with a disease course of 3.24 ± 2.18 years and the latter aged 64.77 ± 10.33 years with a disease course of 4.09 ± 2.63 years. We treated the patients in the control group with Tamsulosin at 0.2 mg qd and those in the combination group with Tamsulosin at 0.2 mg qd plus Xipayimaizipizi at 0.5 g tid, respectively, both for 4 weeks. Then, we obtained the mean frequency of nocturnal urination, maximal urinary flow rate (Qmax), residual urine volume, International Prostate Symptom Score (IPSS) , and quality of life scores (QOL) of the patients, and recorded their adverse reactions.

RESULTSBefore treatment, the nocturnal urination frequency, Qmax, IPSS, and QOL were 3.60 ± 1.81, (10.40 ± 3.53) ml/min, 22.47 ± 8.58, and 4.43 ± 1.50 in the control group, as compared with 3.43 ± 1.61, (10.14 ± 3.43) ml/min, 21.93 ± 8.79, and 4.73 ± 1.31 in the combination group. After 4 weeks of medication, the combination group showed more significant improvement than the control in the nocturnal urination frequency (1.30 ± 1.18 vs 2.27 ± 1.60), Qmax ([13.85 ± 3.15] vs [14.36 ± 3.03] ml/min), IPSS (13.00 ± 1.53 vs 17.20 ± 8.43), and QOL (2.57 ± 1.61 vs 2.93 ± 1.68), all significantly better than the baseline (P < 0.05). The combination therapy achieved remarkable improvement as compared with the control in the nocturnal urination frequency (- [2.13 ± 1.11] vs -[1.73 ± 1.07]), IPSS (- [8.93 ?6.01] vs -[4.80 ± 3.87]), and QOL (- [2.17 ± 1.12] vs -[1.50 ± 1.01]) (P < 0.05), but exhibited no significant differences from the latter in Qmax ([3.72 ± 2.281 vs [3.95 ± 2.53] ml/min) and residual urine volume (- [34.30 ± 37.43] vs - [26.43 ± 30.49] ml) (P > 0.05). Adverse reactions were found in 5 cases in the combination group (16.67%) and 3 cases in the control (10%) , with no remarkable differences between the two groups (P > 0.05).

CONCLUSIONThe combination therapy of Xipayimaizipizi Capsules and Tamsulosin can improve the symptoms of BPH and the patients quality of life of.

Aged ; Capsules ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; drug therapy ; Quality of Life ; Sulfonamides ; therapeutic use

Objective To explore the characteristics of T-cell receptor beta (TCRβ) gene rearrangement in children with T-cell acute lymphoblastic leukemia (T-ALL) and establish a system for quantitative detection of minimal residual disease (MRD) by real-time quantitative PCR (RQ-PCR) targeting the TCRβ gene rearrangement. Methods Multiplex PCR designed by the BIOMED-2 was used to detect TCRβ gene rearrangement in the bone marrow samples of 26 children with T-ALL. Sequences of junctional region were then compared and analyzed in IMGT database. Allele specific oligonucleotide (ASO) upstream primers were designed complementary to the V-D-J or D-J junctional region of TCRβ gene rearrangements. Samples at diagnosis were serially diluted in DNA obtained from mononuclear cells (MNC) from a pool of 20 healthy donors to generate the patient-specific standard curves. Subsequently, TCRβ RQ-PCR was applied to six patients to quantify MRD with germline Jβ primer/probe combinations. To determine the quantity and quality of DNA, we also used RQ-PCR for the N-ras gene.Results Clonal rearrangements were identified in 92.3% of the children with T-ALL ( Vβ-Dβ-Jβ rearrangements in 84.6% and Dβ-Jβ rearrangements in 50% ). Comparative sequence analysis of 42 TCRβ recombination revealed that two downstream Vβ families (BV5, BV6) were preferentially used. The segment Jβ2. 7 was dominant in childhood T-ALL. Jβ1. 3, Jβ2.4, and Jβ2.6 were not detected. The slope of the standard curves was from - 3.54 to -3.37 with the intercepts between 19.35 and 20.51. The correlation coefficients of all the 6 standard curves were ≥0.98. None of the cases had a quantitative range of RQ-PCR lower than 10<'-4>. During the follow-up, an increased incidence of MRD was found before relapse. Conclusions RQ-PCR, which is a highly sensitive and specific method for detection of TCRβ gene rearrangements, will be of high value to study MRD in T-ALL. Close monitoring of MRD is of great importance for prognosis and follow-up of the patients with the disease.

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment

Background The in vitro culture of retinal vascular endothelial cells is the foundation of experimental study of retinal vascular disease. Shortage of human donor eyeballs is a main limiting for the laboratory work. The culture method of rat-derived vascular endothelial cells has been established. However, this method is not enough effective because of severer cellullar injury. Objective Present study was to establish a simple and high effective method for the culture of vascular endothelial cells in vitro. Methods The retinas from 5 SPF SD rats was digested by 0. 1% collagenase and cultured with explant culture method. 20% fetal bovine serum, vascular endothelial growth factor ( VEGF) , insulin-transferrin-selenium( ITS) were composed into the endothelial cell culture medium, and enough blowing was performed to get the cells and fragments from retinal tissue. The cellular suspension was prepared and cultured consequently on human fibronectin-coated culture flasks. Cultured vascular endothelial cells were identified by anti-von Willebrand staining factor. Results The cells emerged from the tissue mass,and cells and some tissue fragments attached to the wall after 24 hours of seeding. The cells grew to show the fusiform in 4 days and merged together in 5 to 6 days,and a cell monolayer was seen in the 14th day after culture. The endothelial cells showed the positive response for von Willebrand factor. After passage, the merging-growth statue of the cells was regained in 2 hours after culture. Conclusion Use of retinal pieces and collagenase-digestion can get the vascular endothelial cells with better activity in vitro. The culture method based on highly selective endothelial cell culture medium associated to FN adhesion-promoting is helpful for gaining the purified of endothelial cells.

BACKGROUNDIgA nephropathy is the major cause of end-stage renal failure in patients with primary glomerular diseases. Tumor suppressor cylindromatosis (CYLD), the recently identified member of the deubiquitinating enzymes, has been actively involved in regulation of inflammation. This study was undertaken to investigate the CYLD expression profile in IgA nephropathy and identify factors associated with CYLD expression.

METHODSForty-one cases of IgA nephropathy were selected. CYLD expression in the kidney biopsy tissue was measured by immunohistochemical staining. Relevant clinical and pathological data were analyzed, and Logistic regression analysis was carried out to identify factors associated with CYLD expression.

RESULTSCYLD was specifically expressed in renal tubular epithelial cells in 70% of the studied patients with IgA nephropathy. All patients with positive CYLD staining had proteinuria, while only 72.7% of patients with negative CYLD had proteinuria (P = 0.003). Among studied proteinuric patients, those with positive CYLD had significantly less tubulo-interstitial lesions and higher estimated glomerular filtration rate (eGFR) levels when compared with those patients showed negative CYLD results. Logistic regression analysis indicated that the urinary protein excretion and eGFR were identified as predictors for the CYLD expression.

CONCLUSIONCYLD is expressed in renal tubular epithelial cells and appears to be associated negatively with tubulointerstitial lesions, however, its exact functional role remains to be clarified in further experiments.

Deubiquitinating Enzyme CYLD ; Glomerular Filtration Rate ; Glomerulonephritis, IGA ; metabolism ; Humans ; Immunohistochemistry ; Kidney Diseases ; metabolism ; Logistic Models ; Proteinuria ; metabolism ; Tumor Suppressor Proteins ; metabolism

Background IgA nephropathy is the major cause of end-stage renal failure in patients with primary glomerular diseases.Tumor suppressor cylindromatosis(CYLD),the recently identified member of the deubiquitinating enzymes,has been actively involved in regulation of inflammation.This study was undertaken to investigate the CYLD expression profile in IgA nephropathy and identify factors associated with CYLD expression.Methods Forty-one cases of IgA nephropathy were selected.CYLD expression in the kidney biopsy tissue was measured by immunohistochemical staining.Relevant clinical and pathological data were analyzed,and Logistic regression analysis was carried out to identify factors associated with CYLD expression.Results CYLD was specifically expressed in renal tubular epithelial cells in 70% of the studied patients with IgA nephropathy.All patients with positive CYLD staining had preteinuria,while only 72.7% of patients with negative CYLD had proteinuria(P=0.003).Among studied proteinuric patients,those with positive CYLD had significantly less tubuto-interstitial lesions and higher estimated glomerular filtration rate(eGFR)levels when compared with those patients showed negative CYLD results.Logistic regression analysis indicated that the urinary protein excretion and eGFR were identified as predictors for the CYLD expression.Conclusion CYLD is expressed in renal tubular epithelial cells and appears to be associated negatively with tubulointerstitial lesions,however,its exact functional role remains to be clarified in further experiments.