Objective To investigate the effects of low-pressure carbon dioxide pneumoperitoneum (CLC) and gasless abdominal distension (GLC) on elderly patients during laparoscopic cholecystectomy. Methods The clinical data,including operation time,hospital stay,surgical complications,and changes of other systems,of 36 elderly patients were analyzed. All the patients were older than 65 years when they received laparoscopic cholecystectomy in our hospital from January 2005 to December 2007. Among the cases,CLC (CO2 pressure:8-10 mm Hg) was used in 24,and GLC was employed in 12. The fourth generation artificial rib lifting apparatus was used for GLC. Results All the procedures were completed by laparoscopy,no patient was converted to open surgery. The operation time of the CLC and GLC groups was (46.2?14.8) min and (52.4?18.6) min respectively (t=-1.087,P=0.285). The vital signs of all the cases were stable during the operation,no carbon dioxide retention or cardiovascular affairs occurred. The hospital stay was (10.5?6.8) d in group CLC and (8.9?5.5) d in group GLC (t=0.706,P=0.485). The patients were followed up for 6 to 42 months,no complications occurred during the period. Conclusions GLC is safe for elderly patients with similar efficacy to that of low-pressure CLC.

An optimized two-dimensional polyacrylamide gel electrophoresis (2-DE) system for analyzing plant proteins was developed by evaluating different reagents and concentrations used in the sample extraction solutions and lysis buffers. Two main sample preparation methods, referred to as trichloroacetic acid (TCA)-acetone method and phenol extraction-ammonium acetate/methanol (phenol-NH4Ac/methanol) precipitation method, were compared. Four ecotypes of reed plants (Phragmites communis Trin.) from the desert region of north-western China were used as experimental materials: (1) swamp reed (SR) which grows in water about 1 m deep; (2) dune reed (DR) which grows on 5～10 m high sand dunes; (3) heavy salt meadow reed (HSMR) which grows on low-lying salt flats; and (4) light salt meadow reed (LSMR) which grows in the transition area between DR and HSMR growing areas. The optimized phenol-NH4Ac/methanol precipitation method consisted of extracting leaf proteins of different ecotypes of reed with water-saturated phenol and then precipitating with a 5-fold volume of 0.1 mol/L NH4Ac in methanol, followed by dissolving in the lysis buffer. The optimized protein lysis buffer consisted of 7 mol/L urea, 2 mol/L thiourea, 4% CHAPS, 2% Ampholine(pH 3.5～10∶pH 5～8 = 1∶4) and 65 mmol/L DTT. The prepared protein sample (80 ?g) was then separated by 2-DE gel and detected by silver staining method. This improved 2-DE system resulted in a 2-D protein profile of higher resolution and higher protein yields as analyzed by PDQuest software. Good results were also obtained when this 2-DE system was used in 2-D analysis of proteins from other plant materials, such as rice leaves, indicating that it is a suitable 2-DE system for analyzing leaf proteins of different plant species.

Objective To explore absorption kinetics of roxatidine acetate hydrochloric (ROX) in intestine of rats.Methods The absorption kinetics and permeability of ROX under different concentrations and different intestinal segments were investigated by double wavelength spectrophotometry via the in situ perfusing method in rats.Results There was no significant difference in Ka of ROX under different concentrations.The absorption rate in rats descended in order of duodenum,jejunum,ileum and colon [(3.87±0.12)×10-2,(2.53±0.18)×10-2,(1.43-±0.10)×10-2,(0.91±0.15)×10-2 · h-1].Conclusion The absorption of ROX in intestine complies with the passive transport mechanism and first order kinetics.ROX is well absorbed in thewhole intestine.

Background Triamcinolone acetonide (TA) is often used in the treatment of uveitis and fundus disease,but whether it has toxic effect to normal tissue around eyes is unclear.Objective The aim of this study was to explore the adverse and toxic effects of TA on the periorbitally normal tissue following the periocular injection.Methods Twenty-seven New Zealand albino rabbits were randomly divided into experimental group,normal saline solution group and blank control group.TA was periocularly injected twice (20 mg for each) at 1-month interval in the rabbits of the experimental group,and 0.5 ml normal saline solution was used in the same way in the rabbits of the normal saline solution group.Not any drug was used in the blank group.The rabbits were sacrificed 1 month and 2 months,3 months after the secondary injection.For the preparation of the specimens of extraocular muscles,peribulbus adipose tissue,lacrimal gland and optic nerve.Hematoxylin and eosin staining of the specimens was performed to examine the the pathological change under the optical microscope,and the ultrastructural of peribulbus tissues was observed under the transmission electron microscope.Results No statistically significant change was found in IOP among various time points and groups (Fgroup =0.952,P =0.881 ; Ftime =7.297,P =0.411).The hyphological structures and ultrastructure of extraocular muscles,peribulbus adipose tissue,lacrimal gland and optic nerve were normal in the rabbits of the normal saline solution group and the blank control group.However,the optical microscopy showed varying degrees of atrophy or dissolving in the extraocular muscle fibers,the increase of glandular epithelial cells and enlargement of nuclei in the lacrimal gland as well as visible disorder arrangement of nerve fibers,vacuoles degeneration and the decrease of glial cells in optial nerve tissue in the rabbits of the experimental group from 1 month though 3 months after the secondary injection.Under the transmission electron microscope,the muscle stripes,bright band and dark band were incompletely disappeared,and expansionary sarcoplasmic reticulum and oval nucleus at the inferior to sarolemma were seen in the extraocular muscles.In addition,swelling lacrimal gland epithelial cells,thickening nuclear membrane and more organelles were exhibited in the lacrimal gland.In the optic never tissue,uneven myelin,space between axons and myelin,lamellar separation and degeneration also were revealed in the rabbits of the experimental group after injection of TA.Peribulbus adipose tissue was near normal both by optical microscope and transmission electron microscope in the experimental group.Conclusions The periocular injection of TA dose not elevated the IOP.However,it will result in the histogical and ultrastructural damages of extraocular muscles,lacrimal gland and optic nerve.

AIM:To further elaborate the effect of glucocorticoid receptor(GR) decrease on level of inflammatory media in rats after critical scald.METHOD:The changes of phospholipase A 2(PLA 2) activity and concentrations of tumor necrosis factor alpha (TNF?) and malondialdehyde (MDA), the product of lipid peroxidation metabolism in plasma and tissue homogenate have been studied in scalded rats with or without GR blockade by RU38486.RESULTS:The PLA 2 activity and the concentrations of TNF? and MDA in plasma and homogenate of pulmonary and renal tissue in scalded rats were significantly higher than those in the controls( P

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 assess the sensitivity and specificity of vaginitis pathogen detection reagent kit (Nucleic acid hybridization). Methods Four hundreds cases of vaginal secretion samples were detected with Amsel, vaginalis culture, fungal culture and Affirm VPIII detection method, respectively. Using the methods of Amsel, vaginalis culture and fungal culture as the gold standard, the clinical application value of Affirm VPIII detection method was evaluated. Results Compared with Amsel, the sensitivity and specificity of Affirm VPIII detection was 92.2%and 70.5%, respectively. Compared with fungal culture, the sensitivity and specificity was 88.3% and 92.9%, respectively. Compared with vaginalis culture, the sensitivity and specificity was 92.6% and 93.8%, respectively. There was a good consistency between the gold stardard and the Affirm VPIII detection. Conclusion Compared with the traditional detection methods,the Affirm VPIII detection has the advantages of fast detection speed,simple operation, and high sensitivity and specificity. In addition, it can identify three kinds of vaginitis pathogenic microorganisms at the same time,with a certain clinical value.

Objective To analyze the relationship between renal pathological characteristics and clinical prognosis in type 2 diabetic kidney disease patients,and discuss predictive value of pathological type and indexes for renal function declining rate and related outcome events.Methods Ninety-two type 2 diabetes patients from PUMC Hospital (with macroalbuminuria and followed up no less than 6 months,excluding patients with non-diabetic renal disease) were divided into typical diabetic glomerulopathy group (DG,n=51) and atypical diabetes-related renal disease group(ADRD,n=41) according to renal pathological findings.A retrospective cohort study was performed to investigate renal pathological features and prognosis.Results Total of 29 renal outcome events and 12 death events occurred in DG group and none in ADRD group; the survival rate and kidney survival rate are different between two groups (P ＜ 0.05); DG group,thick GBM,severe vascular and tubular lesion are predicative indicators for renal outcome event; mesangial volume fraction is predicative indicator for renal outcome events independent of age and serum creatinine.Conclusions DG and ADRD patients have different prognosis and might undergo different pathophysiological mechanisms; renal pathological type and mesangial volume fraction could help predicting outcomes of type 2 diabetic nephropathy patients.

Objective To analyse the operation technique and therapeutic effect of "inserting" uretero-intestinal anastomosis in orthotopic bladder substitution.Methods Thirty-eight patients undergoing orthotopic bladder substitution operations were followed up,and the way of uretero-intestinal anastomosis in all Datients was the "inserting"uretero-intestinal anastomosis.The therapeutic effect was observed by radiation,cystoscopy,pathologic biopsy and blood test.Results The average follow-up time was(3 1.65±14.14)montll8.and the stricture rate was 4%(3/75),but no vesicoureteric reflux was found.The rate of leakage was 0.Nipples were formed at the site of anastomosis under the view of cystoscope,and among the 7 patients whose nipples were taken to be examined by histology,2 cases were intestinal epithelium which were taken at the base of nipple8.while the others were transitional epithelium which were taken at the top of nipples.The renal function of all patients was normal (Cr 54-135 μmol/L,BUN 3.2-9.4 mmol/L).Conclusion "Inserting"uretem-intestinal anastomosis is an ideal antireflux uretero-intestinal anastomosis method.

AIM: To study the effects of [8-(diethylamino) octyl-3, 4, 5 -trimethoxybenzoate] (TMB-8), an intracellular Ca2+ antagonist, on the activation, proliferation and cell-cycle distribution of the mouse T lymphocytes stimulated by concanvalin A (Con A) in vitro. METHODS: After stimulated with Con A, T cells were treated with different concentrations of TMB-8 alone and its combination with cyclosporine A (CsA). The expression of CD69, the early marker of CD3+ T cell activation, was measured by FACS. The proliferation-related index was determined by carboxyl fluorescin diacetate succinmidyl ester (CFDA-SE) flow cytometry. The cell-cycle distribution was analyzed by propidium iodide staining.RESULTS: After 6 h culture, the activation rate of CD69+ T cell in Con A group was (74.88?1.88)%. 10, 20 and 40 ?mol/L of TMB-8 inhibited the expression of CD69 (P