RT-PCR and in situ hybridization were observed during dialyzer reuse. Results Every plasma cytokine level was decreased during reuse compared with first use dialyzer, but no significant difference was found between them. The levels of gene expression of IL-1?、TNF-? and IL-6 were different from the first use significantly. Conclusion If effective dialysis volumn was maintained, formaldehyde as disinfectant on reprocessing the dialyzer may amilorate membrane bio-compatibility. It would be benificial to decrease appearance of long term hemodialysis -related complications.

Objective To investigate the morphological changes of peritoneum during peritoneal dialysis (PD) and elucidate the possible mechanism of its functional deterioration. Methods Peritoneal biopsies were obtained from normal subjects( n = 10), uremic predialysis patients( n = 12) at catheter insertion and PD patients ( n = 10) at the time of catheter remove or reinsertion or renal transplantation, peritoneal morphology was studied by light microscopy, scanning electron microscopy and transmission electron microscopy. Results Normal peritoneal membrane consisted of a monolayer of mesothelial cells on a basement membrane, and a layer of connective tissue containing cells, blood vessels, lymphatic vessels and so on. Mesothelial cells were polygonal, often elongated, and had numerous microvilli on their luminal surface. Sometimes the microvilli ended with roundish formation or resembled a corona. There were lots of oval or roundish pinocytotic vesicles in the cytoplasm of mesothelial cell. Submesothelial connective tissue contained many collagen and elastic fibers. The peritoneal morphology of uremic predialysis patients was similar to that of normal subjects. But significant abnormalities of peritoneal morphology were observed in PD patients and the changes were progressive. Microvilli were the first site of damage, including microvilli shortening, gradual reduction in number and following total disappearance. Then mesolhelial cell detachment from basement membrane and total disappearances were found. Finally the peritoneal membrane only consisted of submesothelial connective tissue denudation of cells. Conclusions PD can modify peritoneal morphology and structure. The morphological change is progressive and might be one of the important causes of peritoneal failure. Peritoneal biopsy can provide lots of valuable informations about the impact of PD, and thus further study on the relationship between peritoneal structure and its function is very useful for understanding of the physiopathology of peritoneum during PD.

Aim To observe changes of gene expression profile in rat heart , liver and brain treated with iptakalim using cDNA microarray and analyze its molecule mechanism about pharmacology and treatment. Methods ①Rats were treated with iptakalim for 14 days at a dose of 3 mg?kg-1 by gavage. Total RNA was extracted from heart, liver and brain tissues and reversely transcribed to cDNA, which were labeled with Cy3 and Cy5 fluorescent as probes and hybridized to Gene Chip (BiostarR-40s). Scan array 3 000 was used for scanning the hybridizing signals and ImageGENE 3.0 for data analysis. Data were confirmed using RT-PCR. ②Rats were randomly divided into normal control group and 4 iptakalim groups. Iptakalim was administered orally at doses of 1, 3, 9 mg?kg-1 body weight per day for 2 weeks. The last group was orally administered at a dose of 9 mg?kg-1 body weight per day for 2 weeks, and iptakalim was withdrawn for 10 days. We observe effects of iptakalim on hemodynamics parameters in anesthetized rats and changes of myocardial structure, myofilament ultra-structure after 24h at the end of the last administration. Results ① The new chemical entity iptakalim had selectivity on changes of gene expression in rat heart, liver and brain. Compared with control group, 236 genes are changed (100 increased and 136 decreased) in rat hearts and 6 transcripts (6 increased) in rat livers, there are no changes on gene expression in rat brains. ② In anesthetized rats, iptakalim at doses of 1, 3, 9 mg?kg-1 neither affected pharmacological effects on cardiovascular hemodynamics parameters nor had pathological changes on myocardial morphological and ultrastructure. Conclusion Under the same experimental conditions, the new chemical entity iptakalim has selectivity on changes of gene expression in vital organs. Iptakalim shows no side-effects on cardiac function and tissue structure.

To determine whether uremia per se can activate peripheral blood mononuclear cells (PBMCs). Methods Maintained hemodialysis patients with cuprophane, non-dialysis uremic patients and healthy volunteers were selected to investigate IL-1?、 TNF-? and 11,6 mRNA in PBMCs by RT-PCR and cells in situ hybridization. Results IL-1?,TNF-? and IL-6 mRNA in PBMCs were undetected in healthy volunteers, but these cytokine gene expression were detected in hemodialysis patients and non-dialysis uremic patients. IL-1?、TNF-? and IL-6 mRNA level was lower in non-dialysis uremic patients than that in hemodialysis patients. Conclusion Uremia per se or its related factors may activated PBMCs and the preactivation of PBMCs might be associated with the body resistance against infection in uremic patients.

Objective To observe the effects of Shenqi Fuzheng injection on peripheral blood T lymphocyte subsets, T-cell apoptosis and cytokine in patients with sepsis and approach their significance.Methods Forty patients with sepsis admitted into Department of General Surgery of the General Hospital of Tianjin Medical University were randomly divided into two groups: routine group (20 cases, received routine western medicine treatment) and Shenqi Fuzheng treatment group (20 cases, received routine western medicine treatment and intravenous drip of Shenqi Fuzheng injection 250 mL per day), 7 days as a course of treatment in both groups. On the 1st, 3rd and 7th day after treatment, evaluation of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores was carried out in the two groups. The percentages of peripheral blood T-lymphocyte subsets CD4+, CD8+, CD4+/CD8+ and apoptosis were measured by flow cytometry. Meanwhile, tumor necrosis factor-α (TNF-α), interleukins (IL-6 and IL-10) levels were detected by enzyme linked immunosorbent assay (ELISA).Results APACHE Ⅱ scores of two groups on the 3rd day were apparently decreased in comparison with those on the 1st day after treatment, and this situation persisted until the 7th day after treatment; meanwhile, the decrease in APACHE Ⅱ score in Shenqi Fuzheng treated group was more notable than that in routine group (10.05±3.71 vs. 13.15±4.65,P < 0.05). Along with the prolongation of time, in both groups, the peripheral blood TNF-α and IL-6 levels were gradually decreased; the IL-10 level was gradually increased, until the 7th day it began to decrease. On the 7th day, the TNF-α and IL-6 levels in Shenqi Fuzheng treated group were decreased more significantly than those in routine group [TNF-α (ng/L): 204.6±18.1 vs. 218.9±21.3, IL-6 (ng/L): 3.68±0.30 vs. 3.95±0.49, bothP < 0.05]. There was no significant difference between Shenqi Fuzheng treated group and routine group in the IL-10 level on the 7th day (ng/L: 173.8±23.3 vs. 174.8±18.9,P > 0.05). On the 1st, 3rd and 7th day after treatment, the percentage of CD4+ T cells and CD4+/CD8+ ratio were firstly fallen and then elevated up, the percentage of CD8+ T cells was gradually decreased, and the percentages of CD4+ and CD8+ apoptosis showed a trend of rise up first and then fall in the two groups. On the 7th day after treatment, there were significant differences between Shenqi Fuzheng treated group and routine group in terms of the percentage of CD4+, CD4+/CD8+ ratio and the rate of CD4+ apoptosis [CD4+ T cells: (38.3±4.7)% vs. (35.5±5.5)%, CD4+/CD8+: 1.55±0.29 vs. 1.36±0.27, CD4+ T apoptosis: (11.2±3.8)% vs. (14.1±5.5)%, allP < 0.05]. There were no statistically significant differences between routine group and Shenqi Fuzheng treated group in the percentages of CD8+ T cells and CD8+ apoptosis at each time point (allP > 0.05).Conclusion Shenqi Fuzheng injection can effectively reduce the percentage of CD4+ T cell apoptosis, increase the percentage of CD4+ T cells and CD4+/CD8+ ratio, lower the inflammatory factors, improve the immune function and disease severity in patients with sepsis.

Objective To compare the effect of different concentrations of ropivacaine in epidural analgesia of childbirth.Methods According to the digital table,300 cases of our hospital childbirth puerpera were randomly divided into A,B,C three groups.A group was given 0.125％ ropivacaine compound fentanyl analgesia,B group was given 0.1％ ropivacaine compound fentanyl analgesia,group C was not required analgesia childbirth.The childbirth of three groups was observed.Results In group A,the labor time was (261.38 ± 19.87) min,postpartum 2h blood loss was (241.03 ± 34.57) mL.In group B,the labor time was (260.09 ± 19.69) mnin,postpartum 2h blood loss was (238.66 ± 35.01) mL.In group C,the labor time was (270.46 ± 20.86) min,postpartum 2h blood loss was (251.75 ± 36.79) mL.Statistical analysis showed that there was no significant difference (t =0.472,1.035 ; all P ＞ 0.05).In group A,the fetal heart rate was (142.34 ±21.57)times/min,neonatal Apgar score was (9.77 ± 0.21),and umbilical artery blood pH value was (7.27 ± 0.06).In group B,fetal heart rate was (145.21 ± 21.49) times/min,neonatal Apgar score was (9.79 ± 0.20),and umbilical artery blood pH value was (7.26 ± 0.08) ; Fetal heart rate of group C was (143.78 ±22.01)times/min,neonatal Apgar score was (9.64 ±0.24),and umbilical artery blood pH value was (7.28 ± 0.07).The differences among three groups were not statistically significant (t =0.763,0.360,0.114,all P＞ 0.05).Analgesic effect time of group A was (12.13 ± 1.76) min,pain score was (1.03 ±0.46) points,analgesic harem duration was (22.39 ± 3.21) s,analgesic harem time interval was (3.26 ± 1.49) min,the cesareandelivery rate was 8.00％.In group B,the analgesic effect time was (12.04 ± 1.69mnin),pain score was (1.01 ± 0.52) points,analgesic harem duration was (21.04 ± 3.18) s,analgesic harem time interval was (3.4.9 ±1.51)min,the cesarean delivery rate was 9.00％ ; Duration of the analgesic effect of group C was (16.77 ±16.77) min,pain score was (3.76 ± 1.23) points,analgesic harem duration was (26.98 ± 5.87) s,analgesic harem time interval was (2.65 ± 0.75) min,the cesarean delivery rate was 48.00％.The differences between groups were statistically significant (chi square or t =6.148,8.522,5.749,4.095,61.316 ;P ＜ 0.05).Conclusion Application of 0.1％ ropivacaine compound fentanyl anesthesia can effectively relieve patients'pain,shorten labor and reduce postpartum 2h blood loss,impact less on contractions at the same time,reduce the incidence of cesarean delivery,has no influence on the neonate,which is worth popularization and application in clinic.

Objective To study the effects of midkine (MK) gene-targeting small interfering RNA (siRNA)on the invasion of melanoma cells.Methods Three MK gene-targeting siRNAs (S1,S2 and S3)were designed,constructed,and transfected into human A375 melanoma cells.Real-time PCR was performed to measure the expression of MK gene and to screen the siRNA with best efficacy.Then,A375 cells were transfected with the optimal siRNA of various doses (3.125,6.25 and 12.5 nmol/L)followed by additional culture of various durations(24,48,72 hours).Some A375 cells remaining untreated served as the blank control group,and some transfected only with liposomes served as the vector control group.Reverse transcription (RT) -PCR and Western blot were conducted to detect the mRNA and protein expression of MK,respectively,MTT assay to observe the adhesion of A375 cells,and Boyden chamber was used to evaluate cell invasion.Results The expression of MK mRNA was downregulated by all the three siRNAs,especially by the siRNA S3,which was used in the following transfection experiment.Real-time quantitative PCR revealed that the MK mRNA expression was reduced by the siRNA in a dose- (r24hours=-0.906,r4Bhours=-0.922,r72hours=-0.939,all P<0.01)and time-dependent(r3.125nmol/L=-0.889,r625nmol/L=-0.935,r125nmol/L=-0.928,all P<0.01)manner.MTT assay showed that the percentage of adhesing cells was 73.66%±2.25%,49.36%±2.16%and 28.35%±1.68%in A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively.The number of cells migrating across the chamber filter was 23.9±1.6,12.1±1.5,5.6±1.2 among A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively,significantly lower than that in the blank control group(36.8±1.5).The percentage of adhesing cells and number of migrating cells decreased with the dose of siRNA(r=-0.936,-0.915,P<0.01,0.05,respectively).Conclusions MK gene might play an important role in the adhesion and invasion of melanonla cells.To down-regulate the expression of MK gene by siRNA may suppress the adhesion and invasion of melanoma cells.

To examine the effects of ischemia and anoxia on cell activation and cell cycle of astrocytes in vitro, the cell cycles and the proliferation of astrocytes in different time points after ischemia and anoxia were studied by flow cytometry and BrdU labeling and the expression of GFAP and cyclin D1 was detected by the fluorescence immunochemistry. After ischemia and anoxia in vitro, the astrocytes in S phase were significantly increased as compared with those in the normal group and the proliferating ability of the astrocytes was highest 6 h after the treatment as revealed by BrdU pulse labeling, but the astrocytes in S phase and proliferating ability were decreased after 6 h. At the early stages of ischemia and anoxia, the positive staining intensity of GFAP was increased, peaked at 6th h, while 12 h after the ischemia and anoxia, the positive staining intensity of GFAP became weak, and the expression of cyclin D1 was gradually increased after the ischemic and anoxic damage. It is concluded that astrocytes are activated to proliferate and enter new cycle events by ischemia and anoxia, and cyclin D1 is implicated in the proliferation and repair of astrocytes. The cell cycle events are closely associated with the proliferation and activation of astrocytes.
Animals, Newborn ; Astrocytes/*cytology ; Cell Cycle ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Cyclin D1/biosynthesis ; Cyclin D1/genetics ; Glial Fibrillary Acidic Protein/*biosynthesis ; Glial Fibrillary Acidic Protein/genetics ; Rats, Wistar

Objective To study the effect of drinking water at different pH on intestinal microbial diversity in SPF mice.Methods Ninety 21-day-old SPF mice were randomly divided into three groups: drinking acidified water (pH 3.0), neutral water (pH 7.0) and alkaline water (pH 9.0), respectively, for 3 months, 30 mice in each group.10 mice were sacrificed at the end of 4, 8 and 12 weeks in each group.Bacteria, virus and parasites in serum were detected according to the national standard for SPF.Then, primer sequences were designed according to the conservativeness of bacteria 16S rRNA, and libraries were constructed.Subsequently, the ileocecal contents were subjected to high-throughput sequencing (Illumina).10 G data were obtained and analyzed.Results The sequencing results revealed that the intestinal microbial abundance of the mice receiving pH 3.0 drinking water was moderate with a high stability.Conclusions Under the conditions of large-scale breeding, the mice receiving pH 3.0 drinking water can maintain the intestinal microbial diversity for a long time, and reach a stable platform.This study provides theoretical and technical support for the stable long-term maintenance of animal quality production.It is also a specific application of gastrointestinal macrogenomics in animal quality control.

Objective To explore the correlation of mycoplasma pneumoniae infection with frequent relapses of steroid-sensitive nephrotic syndrome (SSNS) in children.Methods 35 patients with relapse of SSNS and acute respiratory tract infection were divided into a observation group (mycoplasma pneumoniae infection) and a control group.The clinical and laboratory data including 24 h urine protein (24 h-Upro),urea nitrogen (Bun),serum creatinine (Scr),albumin (Alb) and cholesterol (Chol) were analyzed before and after treatment.Results The clinical and laboratory indexes were obviously improved after treatment,the difference was statistically significant (P ≤ 0.01).24 h-Upro decreased more significantly in the observation group than in the control group after treatment.In the observation group,15 of 18 children achieved the efficacy,9 of whom had complete response and 6 had partial response.In the control group,14 patients achieved the efficacy,6 of whom had complete response and 8 had partial response.Conclusions After treatment,most of the children with frequent relapses of steroid-sensitive nephrotic syndrome induced by cute respiratory infection are relieved.Proteinuria,hypoproteinemia,hyperlipidemia,and renal function were improved in those patients.Therapies with azithromycin achieves a more marked efficacy.