Objective To evaluate the clinical effects and safety of percutaneous nephrolithotomy (PCNL) by middle renal calice used as the main target for the treatment of staghorn stones with the combination of pneumatic and ultrasonic lithotrite.Methods Clinical data of 73 patients underwent PCNL by middle renal calices as main access with 57 incomplete staghorn stones and 35 complete staghorn stones.To observe the situation calculus removal rate and complications.Results Seventy cases (88 sides) underwent one session PCNL by single access tract (middle caliees),3 cases (4 sides) underwent one session PCNL by double access tracts (2 cases by middle and low calices,1 case by up and middle caliees).After the first period of lithoclasty,17 patients (25 sides) residual stones and the stone removal rate 72.8％ (67/92),among these patients,1 case (1 side) had fragments of lateral renal calyeeal stones with no further treatment.Other 16 cases (24 sides)underwent second session PCNL,all were treated by single access tract (middle calices) and 2 cases (2 sides)had extracorporeal shock wave lithotripsy before the second PCNL.After the second period of lithoclasty,76 sides composed of 27 complete staghorn stones and 49 incomplete staghorn stones had no residual fragments with the stone removal rate 82.6％ (76/92).The operative time lasted 120-320 min.Hemoglobin dropped 1-4 g/L,11 cases in the operation procedure and 3 cases after operation needed blood transfusion respectively.One case of renal pelvic infection after operation and 1 case had split renal dysfunction with peri-parenchyma infection.The hospitalization time was 9-18 days.Conclusion It is effective and safe to perform PCNL for staghorn stone by middle calices as a main access.Combining pneumatic and ultrasonic lithotrite will be very useful with high stone clearance,short procedure time and less complications.

Objective To discuss the medium-term follow-up of clinically insignificant residual fragments (CIRF) after minimally invasive percutaneous nephrolithotomy lithotripsy (MPCNL).Methods The clinical data of 72 patients with CIRF medium-term follow-up were analyzed retrospectively.Results Seventy-two patients with CIRF.The anatomical distribution of CIRF was 10 at upper pole,15 at middle,35 at lower,10 at renal ureteropelvie junction and 2 at upper and lower pole.Stone analysis showed that 41 cases of calcium oxalate calculi,16 of calcium oxalate calculi mixed with carbonate calculi,3 calcium oxalate calculi mixed with uric acid,4 calcium oxalate calculi mixed with struvite stone,3 struvite stone,2 uric acid stone and 3 carbonate apatite mixed with struvite stone.Fifteen cases had clinical symptoms,including 2 renal colic pain,8 hematuria,5 lower urinary tract symptoms,4 cases CIRF located in upper pole,1 case in middle pole,4 cases in lower pole,6 cases in ureteropelvic junction,the incidence of clinical symptoms in ureteropelvic junction was significantly higher than that in other locations (6/10 vs.4/12,1/15,4/37,P ＜0.05).Eight cases required surgical procedure,5 cases underwent extracorporeal shock wave lithotripsy,3 cases with ureteral CIRF were performed with ureteroscopic lithotripsy.CIRF were clear after surgery,7 patients with ureteral CIRF had renal colic pains.The stones were excluded after spasmolytic analgesic treatments.Conclusions CIRF can be located variously in the kidney and ureter.Most CIRF are calcium oxalate calculi and locate in the lower pole.Patients with the history of previous open surgery or extracorporeal shock wave lithotripsy are more likely to get CIRF.Medium-term follow-up of CIRF reveals that CIRF located in the renal ureteropelvis junction are more likely to have clinical symptoms.

Objective To investigate the therapeutics of solitary kidney complicated with complexcalculi,and improve the effect and safety of treatment.Methods Experiences in the treatment of 32 patients with solitary kidney complicated with complex calculi were summarized.Congenital solitary kidney was 6 cases (18.8%),postnatal reason was 26 cases(81.2%),left was 12 cases(37.5%),right was 20 cases (62.5%).All patients were with mould or multiple calculi,9 cases were complicated with ureter calculi,and 8 cases were hospitalized because of obstructive anuria.The patients with mould calculi received extracorporeal shock-wave lithotripsy (ESWL) prior to percutaneous nephrolithotomy(PCNL).While the patients with multiple calculi received PCNL prior to ESWL. Some cases were treated by lithedialysis.Results Twenty-nine cases (90.6%)were cured by ESWL combined with PCNL 12 cases received lithodialysis during PCNL. Eight cases with obstructive anuria recovered in 12 hours after emergent ESWL or lithodialysis,3 cases(9.4%)underwent open operation because of deformity or obstruction in renal pelvis and ureter,1 case had to keep nephrostomy because of repeated infection.Followed up 4-36 months,29 cases (90.6%)kept good kidney function,3 cases(9.4%)had renal insufficiency,2 cases(6.2%)reoccurred calculi.Conclusions The therapeutics of ESWL combined with PCNL may clear complex calculi of solitary kidney effectively and safely.It is necessary to take emergent ESWL in renal obstructive calculi cases.And the patients with lower ureter obstructive calculi may take lithodialysis first.It is proper to choose open operation on the patients with deformity of renal pelvis or obstruction of ureter.

Objective To compare the outcomes of antegrade and retrograde approach ureteroscopy for impacted upper ureteric calculi and assess the safety and efficiency of the two types of minimally invasive technique. Methods A total of 106 patients with impacted upper ureteric calculi were treated with ureteroscopy. The procedure was performed via antegrade percutaneous nephrostomy tract in 50 patients (antegrade group) and via retrograde transurethral access in 56 patients (retrograde group). Results The success rate of retrograde group was 92.9% (52/56). Operating time was (45 ± 5 ) min, hospital stay was (6 ± 1) days. The stone free rate was 80.4%(45/56) at 1 month follow-up,7 patients with residual calculi required ESWL combination. Complication rate was 5.4% (3/56). The success rate of antegrade group was 100.0% (50/50). Operating time was (55 ± 8 ) min, hospital stay was (8 ± 2) days. The stone free rate was 100.0% (50/50) and no complication was noted. The stone free rate and the complication rate indicated significant difference between the two groups (P ＜ 0.05). Conclusions Antegrade and retrograde access ureteroscopy for impacted upper ureteric calculi are safe and effective. Success rate and stone free rate of antegrade approach are higher than those of retrograde approach.

BACKGROUND:At present, the separation and culture technique of chondrocytes has been mature, but the chondrocytes grow slowly which are prone to degenerate using the present technique. It is not conducive to the fol ow-up test.
OBJECTIVE:To investigate and improve the separation and culture method of articular chondrocytes of New Zealand rats at 4 weeks of age.
METHODS:New Zealand rats aged 4 weeks were selected to take cartilage tissues from the bilateral knees that were resected under aseptic condition. Chondrocytes were isolated by type Ⅱ col agenase enzyme digestion and mechanical isolation method. The cells were cultured and passaged, and then identified by morphologic observation, toluidine blue staining and type Ⅱ col agen enzyme immunohistochemical methods. Growth curve was pictured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.
RESULTS AND CONCLUSION:Inverted microscope observation showed that the primary cultured chondrocytes adhered at 6 hours after cultivation. The monolayer formation occurred at 72 hours after cultivation, and the cells were ready to be passaged at 96 hours after cultivation. In the fourth generation, some cells represented a spindle-like appearance. In the fifth generation, most cells turned into irregular shape appearance, and cellproliferation capacity diminished. Toluidine blue staining showed that the nuclei of cultured chondrocytes were blue and cytoplasm was pale blue. Immunofluorescent staining showed that cultured chondrocytes had a positive expression of col agen type Ⅱ and the color was tawny. Proliferative rate of chondrocytes in the first to third generations had no differences (P<0.05), while differences were found compared with the fourth generation in 4-7 days (P<0.05) and the fifth generation in 1-7 days (P<0.05). The results indicate that type Ⅱ col agenase enzyme digestion and mechanical isolation method is successful for isolating, cultivating New Zealand rat articular chondrocytes in vitro, and the first to third generations can be the best choice for the experiments of knee osteoarthritis.

Objective　To study the transcription effects and cleavage activities of rat caspase-3 specific hammerhead ribozyme (Rz107) in both cell-free conditions and BRL-3A cells. Methods　Rat caspase-3 gene fragment was cloned into the pGEM-T EASY vector under the T7 promoter control. The 32　 P-labeled caspase-3 transcript was the target-RNA. Rz107 genes designed against caspase-3 mRNA were cloned into vector p1.5 between 5'-cis-Rz and 3'-cis-Rz. 32　 P-labeled ribozyme transcripts were incubated with target-RNAs at different conditions and autoradiographed after denaturing gel-electrophoresis. Rz107 was electroporated into BRL-3A cells and the Rz107 expression was analyzed by RT-PCR. Results　In cell-free conditions, Rz107 was active at 37℃. The optimal temperature was 50℃. The Km and kcat were 14.13?nmol/L and 2.31*min-1 respectively. Intracellular cleavage efficiency of Rz107 was 37%, as analyzed by RT-PCR. This indicated that the design of Rz107 was correct, and Rz107 had the activity of common enzymes. Conclusions　Rz107 in cell-free conditions possessed perfect specific catalytic cleavage activity, and it can also cleave the target RNA successfully in cells. The results illustrate the feasibility of ribozyme therapy as a potential alternative approach for treating liver disease caused by apoptosis.

A Marfan syndrome child with contractural arachnodactyly deformity of both hands was admitted to our unit in June 2015. The left and right middle fingers and ring fingers of the child received the continuous " Z" skin flap and " V-Y" plasty to reduce the palmar tension on the skin and release the contracture. Due to the improper therapeutic strategy and the underestimation of effect of underlying lesions of Marfan syndrome on the therapeutic strategy, the right middle finger of child was with ischemic necrosis and amputated, and the finger tip was accompanied with distension and paroxysmal neuropathic pain at a specified future date. The skin flaps of the other three surgical fingers were with local necrosis and scar healing. The child was complicated by distal interphalangeal joints stiffness and some serious complications post-discharge. Combined with this case, the similar disorders are not recommended to carry out one-stage operation.

Objective To explore the microRNA-16 (miR-16) and nuclear-transcription factor-κB1 (NF-κB1) expressions in human brain gliomas and their correlations with cell invasion and growth of malignant gliomas SHG44,U87 and U373.Methods (1) Twenty-nine cases of human glioma tissue samples and 6 normal brain tissues,collected in our hospital from January 2000 to January 2011,were chosen in our study; quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the expressions ofmiR-16 and NF-κB1 in these tissues.(2) In vitro cultured U87,U373 and SHG44 cells were divided into blank-control group,nonsense sequence transfected group and miR-16 mimics transfected group; 48 h after the transfection,qRT-PCR was used to detect the expressions of miR-16 and NF-κB1; tmnswell assay was used to observe the cell invasion capability; 72 h after the transfection,Western blotting was employed to detect the protein expressions of NF-κB1,matrix metallopeptidase 9 (MMP-9) and MMP-2.(3) Luciferase reporter assay was used to detect the target regulating role of miR-16 in NF-κB1 gene.(4) U87 cells were used as negative control group,and U87 cells carried stably expressed miR-16 gene were implanted into intracranial and subcutaneous nude mice (U87-miR-16 group); immunofluorescence was used to detect the MMP-9 expression,and immunohistochemical staining was used to detect the protein expressions of Ki-67,NF-κ B1 and MMP-9; subcutaneous tumor volume was measured and the growth curve was drawn.Results (1) The qPCR results showed that the expression of miR-16 in human brain glioma tissues was significantly lower than that in normal brain tissues; and gradually decreased miR-16 expressions were noted in gliomas of graded Ⅰ,Ⅱ,Ⅲ and Ⅳ (P＜0.05); NF-κB1 expression in human brain glioma tissues was significantly higher than that in normal brain tissues; and gradually increased NF-κB1 expressions were noted in gliomas of graded Ⅰ,Ⅱ,Ⅲ and Ⅳ (P＜0.05).(2) As compared with those in the blank-control group and nonsense sequence transfected group,miR-16 mimics transfected group had significantly increased miR-16 expression,decreased NF-κB1 mRNA expression,decreased invasiveness,and decreased protein expressions of NF-κB1 and MMP-9 (P＜0.05).(3) Luciferase reporter assay showed that the fluorescence normalized ratio in the pMIR-NF-κB1 group was signfcaintly higher than that in the pMIR-NF-κB1+pre-miR-16 group (P＜0.05).(4) As compared with the negative control group,the U87-miR-16 group on the 24-42 d of implantation had significantly smaller volume of tumors (P＜0.05),and lower MMP9 expression,and NF-κB1,MMP-9 and Ki-67 expressions (the proliferation index of Ki-67:13.91％ and 32.98％).Conclusion MiR-16 inhibits glioma cell invasion and growth through down-mgulating NF-κB1 and MMP-9 expressions.

OBJECTIVETo establish a new glioma cell line and analyze its biological characteristics, and to provide a useful cellular tool with new features for cancer research.

METHODSGlioma tissue was taken from surgical specimen clinical of a clinical patient. Primary culture was carried out, and a cell line (SHG139) was established after 10 passages. Immunofluorescence staining was performed to detect the expression of proteins, and cell proliferation and cycle were detected by flow cytometry method (FCM). The biological characteristics of SHG139 cells were detected by chromosome karyotype analysis. SHG139s glioma cells derived from SHG139 glioma cell line were cultured with neural stem cell medium. Then stem cell markers were determined. SHG139s cells were induced with serum-containing medium, and their expression of A2B5, GFAP, β-III tubulin, and GalC was detected. Intracranial xenograft tumor of both SHG139 glioma cells and SHG139s glioma stem cell spheres was generated in rats.

RESULTSThe expressions of A2B5, GalC, GFAP, S-100, and vimentin in the 20 and 60 passages of SHG139 cells were positive, consistent with the immunohistochemical results and pathological features. SHG139 cells proliferated significantly within 24 h after subculture, and their total number of chromosomes was 68 and mostly multiploid. They were positive for A2B5 (84.12±9.96)%, nestin (73.86±5.01)%, and NG2 (73.37±2.09)%. SHG139s cells were induced, and the ratio of positive cells of GFAP, β-III tubulin and GalC was (92.89±2.24)%, (64.85±4.09)% and (33.57±4.14)%, respectively.

CONCLUSIONSSHG139 is an astroglioma cell line, from which SHG139s cells can be successfully obtained by culture with NSCM. SHG139s cells are of A2B5(+)/CD133(-) GSCs subgroup cells, with potentials of self-renewal and multi-directional differentiation. Compared with the intracranial SHG139 xenograft tumor, the intracranial SHG139s xenograft tumor is more malignant and aggressive.

Animals ; Astrocytoma ; Brain Neoplasms ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Glioma ; Humans ; Neoplastic Stem Cells ; Nestin ; Rats ; Transplantation, Heterologous ; Vimentin

Objective:To compare the therapeutic effect of lateral position and half lithotomy position in Asian proximal femur intramedullary nail antirotation system (PFNA-II) for treating the elderly patients with femoral intertrochanteric fractures.Methods:A retrospective case control study was made on 141 patients with femoral intertrochanteric fractures admitted to Jiangjin Central Hospital from January 2016 to September 2017, including 54 males and 87 females, aged 65-99 years (mean, 80.4 years). According to AO classification, there were 42 patients with type A1 fractures, 88 with type A2 and 11 with type A3. Of all, 74 patients were stabilized by PFNA-II internal fixation in lateral position (lateral position group) and 67 patients by PFNA-II internal fixation in half lithotomy position (half lithotomy position group). The postural placement time, total incision length, operative time, intraoperative blood loss, fluoroscopy frequency, tip-apex distance, reduction quality, fracture healing time, postoperative complications and Harris hip function at 12 months after surgery were compared between the two groups.Results:All patients were followed up for 12-18 months (mean, 12.5 months), except that 13 patients were lost after 9 months, an average of 12.5 months. There were no statistically significant differences in postural placement time, operative time, fracture healing time, and Harris hip score between the two groups ( P>0.05). While significant differences were seen between lateral position group and half lithotomy position group regarding the incision length [(6.5±1.3)cm vs. (7.5±1.5)cm], intraoperative blood loss [(84.3±3.1)ml vs. (90.4±3.9)ml], fluoroscopy frequency [(13.1±1.9)times vs. (11.2±1.2)times], tip-apex distance [(20. 6±2.2)mm vs. (24.4±1.8)mm], good rate of reduction quality (80% vs. 85%) and implant related complications (5% vs. 2%) ( P<0.05 or 0.01). Conclusion:For treatment of elderly patients with intertrochanteric fractures, compared to the lateral position, the half lithotomy position in PFNA-II internal fixation can reduce frequency of fluoroscopy, improve quality of fracture reduction and reduce implant-related complications.