AIM: To explore the feasibility of culturing human umbilical vein endothelial cells ( HUVEC ) on acellular corneal stroma and performing the posterior lamellar endothelial keratoplasty ( PLEK ) treating corneal endothelial decompensation.
METHODS: Thirty New-Zealand rabbits were divided into three groups randomly, 10 rabbits for experimental group, 10 for stroma group and 10 for control group. Corneal endothelial cells were removed to establish animal model of corneal endothelial failure. PLEK was performed on the rabbits of experimental group and stroma group, and nothing was transplantated onto the rabbits of control group with the deep layer excised only. Postoperative observation was taken for 3mo. The degree of corneal edema and central corneal thickness were recorded for statistical analysis.
RESULTS:Corneas in experimental group were relieved in edema obviously compared with that in stroma group and the control group, and showed increased transparency 7d after the operation. The average density of endothelial cells was 2 026. 4±129. 3cells/mm2 , and average central corneal thickness was 505. 2±25. 4μm in experimental group, while 1 535. 6±114. 5μm in stroma group and 1 493. 5±70. 2μm in control group 3mo after operation.
CONCLUSION:We achieved preliminary success in our study that culturing HUVEC on acellular corneal stroma and performing PLEK for corneal endothelial decompensation. HUVEC transplanted could survive in vivo, and have normal biological function of keeping cornea transparent. This study provides a new idea and a new way clinically for the treatment of corneal endothelial diseases.

Objective To establish the m ethod to analyze γ-hydroxybutyric acid (GHB ) and its precursors 1,4-butanediol (1,4-B D ) and gam m a-butyrolactone (GBL) in urine through LC-M S/M S and provide evi-dence for related cases. Methods GHB-d6 and M O R-d3 were used as the internal standard. The urine sam ple w as separated by LC after protein precipitation w ith m ethanol. The electrospray ion source w as for ionization. E ach com pound w as detected through m ultiple-reaction m onitoring (M R M ) m ode. Results The lim its of detection of GHB and its precursors 1,4-B D and GBLwere 0.1, 0.1 and 2μg/m L. The accuracy w as 87.6% -98.1% . The intra-day and inter-day precisions were less than 15% and m atrix ef-fects were higher than 80% . Conclusion The m ethod is high sensitive, sim ple, rapid, specific and w ith high reliability. This study has provided technical support and basic data for forensic cases involving GHB .

Objective To investigate the clinical value and adverse reactions of indwelling ureteral stentsinthepreventionofureteralinjurybeforecomplexpelvictumorsurgery.Methods 145casesofpatients with rectal cancer ,cervical cancer ,ovarian cancer and pelvic sarcomas were retrospectively analyzed ,and 53 pa-tients with complex pelvic tumor surgery ,preoperative were under cystoscope unilateral or bilateral ureteral stent tube,pulled out according to the intraoperative situation after surgery or lien ,92 patients as control group .Results Ureteral injury was found in 10 of the 145 patients,2 cases in ureteral catheter group and 8 cases in control group .3 cases of postoperative ureteral fistula occurred in the control group .Indwelling ureteral stents could cause adverse reactions such as hematuria ,osphyalgia and urinary irritation ,and the adverse reactions of catheter group was obviously higher than that of control group (P<0.05).Conclusion Cystoscopic ureteral stent placement has important clinical significance for prevention of ureteral injury despite certain adverse reactions ,which can be used in operation of complex pelvic tumor .

OBJECTIVE: To establish the method to analyze γ-hydroxybutyric acid (GHB) and its precursors 1,4-butanediol (1,4-BD) and gamma-butyrolactone (GBL) in urine through LC-MS/MS and provide evidence for related cases. METHODS: GHB-d6 and MOR-d3 were used as the internal standard. The urine sample was separated by LC after protein precipitation with methanol. The electrospray ion source was for ionization. Each compound was detected through multiple-reaction monitoring (MRM) mode. RESULTS: The limits of detection of GHB and its precursors 1,4-BD and GBL were 0.1, 0.1 and 2 μg/mL. The accuracy was 87.6%-98.1%. The intra-day and inter-day precisions were less than 15% and matrix effects were higher than 80%. CONCLUSION The method is high sensitive, simple, rapid, specific and with high reliability. This study has provided technical support and basic data for forensic cases involving GHB.
4-Butyrolactone/urine* ; Butylene Glycols/urine* ; Chromatography, Liquid ; Forensic Sciences ; Humans ; Hydroxybutyrates/urine* ; Mass Spectrometry ; Reproducibility of Results ; Tandem Mass Spectrometry

Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.
Animals ; Cations ; Cell Line ; DNA ; Genetic Vectors ; Kidney ; cytology ; Liposomes ; Magnetics ; Nanoparticles ; Swine ; Transfection

The design space of the high shear wet granulation process was established and validated within the framework of quality by design (QbD). The system of microcrystalline cellulose-de-ioned water was used in this study. The median granule size and bulk density of granules were identified as critical quality attributes. Plackeet-Burmann experimental design was used to screen these factors as follows: dry mixing time, the impeller and chopper speed of dry mixing, water amount, water addition time, wet massing time, the impeller and chopper speed of wet massing and drying time. And the optimization was implemented with the central composite experimental design based on screened critical process parameters. The design space of the high shear wet granulation process was established based on the quadratic polynomial regression model. Since the P-values of both models were less than 0.05 and values of lack of fit were more than 0.1, the relationship between critical quality attributes and critical process parameters could be well described by the two models. The reliability of design space, illustrated by overlay plot, was improved with the addition of 95% confidence interval. For those granules whose process parameters were in the design space, the granule size could be controlled within 250 to 355 μm, and the bulk density could be controlled within a range of 0.4 to 0.6 g x cm(-3). The robustness and flexibility of the high shear wet granulation process have been enhanced via the establishment of the design space based on the QbD concept.
Cellulose ; chemistry ; Reproducibility of Results ; Technology, Pharmaceutical ; methods ; Water

Objectlve To research and analyze the hemod)rnamic status of refractory septic shock associated cardiac dysfunction.Methods 70 refractory septic shock patients were studied.In the duration of pulmonary artery catheter(PAC)-directed hemodynamic optimization,the patients were divided into a cardiac dysfunction group and a control group.Hemodynamic parameters,arterial blood lactate concentration and APACHE II scores were obtained instantly after the placement of a PAC,then lactate clearance in 24 hours was surveyed and calculated.Subsequently the two groups of patients were regrouped by nonsurvivor and survivors respectively.All the obtained values were analyzed with statistic methods.Results 37% of the refractory septic shock patients was complicated with cardiac dysfunction.The age of the patients complicated with cardiac dysfunction was significantly higher than that of the patients of the control group.Central venous pressure(CVP),pulmonary artery obstruction pressure(PAOP),pulmonary artery pressure (PAP),systemic vascular resistance index(SVRI),pulmonary vascular resistance index(PVRI)and oxygen extraction ratio(O2ext)in the cardiac dysfunction group were significantly different from those in the control group.Cardiac output(CO),cardiac index(CI),oxygen delivery index(DO2I)and mixed venous oxygensaturation(S-v O2)were significantly lower than those of the patients in the control group.S -v O2 had a strong correlation witIl CI.If the patients were regrouped by nonsurvivors and survivors.in the patients complicated with cardiac dysfunction APACHE II scores were significantly higher in the nonsurvivors than survivors:the lactate clearance in 24 hours(median-25%)of the nonsurvivors was significantly lower than that of nonresponders(median 22%),P＜0.05.Conclusion (1)In refractory septic shock patients,cardiac dysfunction maybe the main reason leading to bad outcome.(2)Higher CVP and PAOP and lower S -v O2 indicate the onset of cardiac dysfunction.(3)The patients with significantly high initial arterial blood lactate level and the low lactate clearance in 24 hours had bad outcome.

Objective To investigate the clinical role of central venous pressure(CVP) to evaluate fluid responsiveness in septic shock patients. Methods 66 septic shock patients were studied, every patient was administered a volume challenge, before and after it, CVP, intrathoracic blood volume index (ITBVI),global end-diastolic volume index(GEDVI), cardiac index(CI), stroke volume index(SVI) were measured by PiCCO method. All the obtained values were analyzed by statistics method. Results Initial CVP in responders is significantly different from that in nonresponders; △ITBVI, △GEDVI, △CI, △SVI, △HR (△:changes) before and after volume challenge in responders were significantly different from those in nonresponders; the significance of △ITBVI, AGEDVI to predict volume responsiveness was strong indicated by high values of areas under the receiver operating characteristic curves (0.674 and 0.700, respectively).If patients were regrouped by CVP≤11 mm Hg(1 mm Hg=0.133 kPa) and CVP > 11 mm Hg, initial ITBVI and GEDVI in responders were not significantly different from that in nonresponders; △ITBVI,△GEDVI, △CI, △SVI before and after volume challenge in responders were significantly different from those in nom'esponders. Conclusion In septic shock patients, CVP play a guidance role to predict and evaluate volume responsiveness and when CVP was > 11 nun Hg, a positive response will be less likely. Initial volumetric parameters(intrathoracic blood volume and global end-diastolic volume) play a questionable role in predicting and evaluating volume responsiveness, changes before and after volume challenge maybe helpful.

Objective To explore the influence of hemodialysis(HD) and henodiafiltation on serum ?_2-microglobulin(?_2-MG) of children with acute renal failure and contrast research on declining rate of serum ?_2-MG before and after(children′s) HD and HDF.(Met-)hods By Branc Dialog HD machine of double-pump,HDF for 18 times and HD for 20 times were given to children with acute renal(fai-)lure.The serum ?_2-MG were observed before and after HD and HDF.Results A great difference was observed in level of serum ?_2-MG between before and after HD and HDF.In HDF group,there was significant difference in level of serum ?_2-MG before and after HDF(P

Objective: To determine the glycemic index of common cereals and tubers products in China, and to examine the relationship among the response of blood glucose and the type of carbohydrate, food processing, and food digestion and absorption. Methods: 8-12 subjects in each group were assigned randomly. Fasting blood sugar was measured first and then 50 g glucose or the test meal was taken, and blood glucose was measured again 2 hours later.The food used in the test meal contained the carbohydrate content, equivalent to 50 g glucose according to the Table of Food Composition (1991). 50 g glucose was used as the control food. GI of test meal was calculated by Wolver method. Results:The study showed the glycemic index of common foods, including 9 sugars, 62 cereals and tubers products. Conclusion: The different foods with same amount of carbohydrate have different GI. The characteristics of starch and food processing are more important in predicting GI value, and GI varies also with the rate of starch digestion and hydrolysis in man.