The clinical study on the relationship between serum albumin concentration and lymphocyte levels in patients with 2019-novel coronavirus pneumonia / 中华急诊医学杂志

Objective@#To explore the relationship between different serum albumin and lymphocyte levels in patients with 2019-novel coronavirus (2019-nCoV) pneumonia (COVID-19).@*Methods@#A retrospective study was performed to identify the characteristics of the clinical data of 205 COVID-19 patients who were hospitalized in the Happy Street of Hanchuan People's Hospital, Xiaogan, Hubei Province from January 24 to February 12, 2020, including their general information, serum albumin (ALB) levels, lymphocyte counts (LYM), percentage of lymphocytes (LYM%) and other laboratory parameter levels. Low ALB group and normal ALB group were demarcated by the concentration of 35g/L, further to identify the differences of LYM and LYM% levels and the incidence of LYM and LYM% decline at different ALB levels between groups,as well as the correlation between ALB and LYM, LYM% levels in hypoalbuminemia conditions .@*Results@#17.5% of COVID-19 patients were associated with hypoalbuminemia. The levels of LYM and LYM% in the low ALB group were significantly lower than those in the normal ALB group (P <0.001). The incidence of LYM and LYM% decline in the low ALB group was significantly higher than those in the normal ALB group (P <0.001). The levels of LYM and LYM% in the low ALB group were significantly positively correlated with serum ALB concentrations (P <0.05).@*Conclusions@#The decrease of lymphocyte levels in COVID-19 patients may be correlated to hypoalbuminemia. COVID-19 patients complicated by hypoalbuminemia should be actively intervened to maintain serum albumin in the normal range.

The clinical study on the relationship between serum albumin concentration and lymphocyte levels in patients with 2019-novel coronavirus pneumonia / 中华急诊医学杂志

Objective:To explore the relationship between different serum albumin and lymphocyte levels in patients with 2019-novel coronavirus (2019-nCoV) pneumonia (COVID-19).Methods:A retrospective study was performed to identify the characteristics of the clinical data of 205 COVID-19 patients who were hospitalized in the Happy Street of Hanchuan People's Hospital, Xiaogan, Hubei Province from January 24 to February 12, 2020, including their general information, serum albumin (ALB) levels, lymphocyte counts (LYM), percentage of lymphocytes (LYM%) and other laboratory parameter levels. Low ALB group and normal ALB group were demarcated by the concentration of 35 g/L, further to identify the differences of LYM and LYM% levels and the incidence of LYM and LYM% decline at different ALB levels between groups,as well as the correlation between ALB and LYM, LYM% levels in hypoalbuminemia conditions .Results:17.5% of COVID-19 patients were associated with hypoalbuminemia. The levels of LYM and LYM% in the low ALB group were significantly lower than those in the normal ALB group ( P <0.001). The incidence of LYM and LYM% decline in the low ALB group was significantly higher than those in the normal ALB group ( P <0.001). The levels of LYM and LYM% in the low ALB group were significantly positively correlated with serum ALB concentrations ( P <0.05). Conclusions:The decrease of lymphocyte levels in COVID-19 patients may be correlated to hypoalbuminemia. COVID-19 patients complicated by hypoalbuminemia should be actively intervened to maintain serum albumin in the normal range.

[Morphometric evaluation of changes in the alveolar bone of adolescents with bimaxillary protrusion via cone beam computed tomography].

OBJECTIVE: This study aimed to evaluate the morphometric changes in the alveolar bone of the maxillary and mandibular anterior regions after retraction in adolescents. METHODS: The sample size comprised 30 adolescent patients with class 1 bimaxillary protrusion (12 males and 18 females, age: 12-18 years old) and were treated by extracting four first pre-molars. Cone beam computed tomography (CBCT) was performed 1 month before and 1 month after the retraction. For each maxillary and mandibular anterior tooth, the labial and palatal alveolar plates at cervical 1/3, middle 1/3, and apical 1/3 levels for bone thickness changes during the retraction of the maxillary and mandibular anterior regions were checked. The movements of cervical 1/3, middle 1/3, and apical 1/3 levels of the maxillary central incisor were measured. Statistical analyses were performed with SPSS 16.0. RESULTS: For the adolescents, alveolar bone thickness increased on the labial side and decreased on the palatal side. The alveolar bone thicknesses of cervical 1/3 and middle 1/3 of maxillary central incisor, cervical 1/3 and apical 1/3 of maxillary lateral incisor, middle 1/3 of mandibular central incisor, apical 1/3 of mandibular lateral incisor, and middle 1/3 and apical 1/3 of mandibular canine all increased after retraction. By contrast, the alveolar bone thickness of the apical 1/3 of maxillary canine and the cervical 1/3 of mandibular canine decreased after retraction. No statistically significant difference was observed in other region. CONCLUSION: During retraction, a controlled tipping movement occur in adolescents. After retraction, the alveolar bone thickness of the labial side increase, whereas that of the palatal side decrease. Moreover, the thicknesses of major areas in the alveolar bone significantly increase.

Morphometric evaluation of changes in the alveolar bone of adolescents with bimaxillary protrusion via cone beam computed tomography / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aimed to evaluate the morphometric changes in the alveolar bone of the maxillary and mandibular anterior regions after retraction in adolescents.</p><p><b>METHODS</b>The sample size comprised 30 adolescent patients with class 1 bimaxillary protrusion (12 males and 18 females, age: 12-18 years old) and were treated by extracting four first pre-molars. Cone beam computed tomography (CBCT) was performed 1 month before and 1 month after the retraction. For each maxillary and mandibular anterior tooth, the labial and palatal alveolar plates at cervical 1/3, middle 1/3, and apical 1/3 levels for bone thickness changes during the retraction of the maxillary and mandibular anterior regions were checked. The movements of cervical 1/3, middle 1/3, and apical 1/3 levels of the maxillary central incisor were measured. Statistical analyses were performed with SPSS 16.0.</p><p><b>RESULTS</b>For the adolescents, alveolar bone thickness increased on the labial side and decreased on the palatal side. The alveolar bone thicknesses of cervical 1/3 and middle 1/3 of maxillary central incisor, cervical 1/3 and apical 1/3 of maxillary lateral incisor, middle 1/3 of mandibular central incisor, apical 1/3 of mandibular lateral incisor, and middle 1/3 and apical 1/3 of mandibular canine all increased after retraction. By contrast, the alveolar bone thickness of the apical 1/3 of maxillary canine and the cervical 1/3 of mandibular canine decreased after retraction. No statistically significant difference was observed in other region.</p><p><b>CONCLUSION</b>During retraction, a controlled tipping movement occur in adolescents. After retraction, the alveolar bone thickness of the labial side increase, whereas that of the palatal side decrease. Moreover, the thicknesses of major areas in the alveolar bone significantly increase.</p>

Real-time ultrasound guided placement of permanent internal jugular vein catheters in maintenance hemodialysis patients / 中南大学学报(医学版)

OBJECTIVE@#To investigate the value of real-time ultrasound guided placement of permanent internal jugular vein (IJV) catheterization in maintenance hemodialysis patients, and analyze its technical success and complication rate.@*METHODS@#We prospectively analyzed 63 patients (39 males, 24 females) who underwent permanent IJV cannulation with real-time ultrasound guidance from January to October in 2012. Under the real-time guidance of Logiq 5 color Doppler, we placed the tunneled cuffed catheters into the jugular vein by Seldinger technique. The number of needle punctures, technical success, the operation time, and complications were recorded. The patients were divided into a normal-risk group and a high-risk group: those who suffered multiple catheter insertions, previous difficulties during catheterization, poor compliance, obesity, impaired consciousness, skeletal deformity, disorder of haemostasis were regarded as high-risk patients.@*RESULTS@#Cannulation of IJV was done in all patients. Of the 63 catheters, 20 (31.7%) were placed in the high-risk patients; 60 (95.2%) were successfully placed at the first attempt, with the average number of punctures of (1.23±0.21) (range 1-3); Only 3 immediate complications (4.7%) developed; 3 (4.7%) catheter infections occurred in the course of using. Cannulation of IJV took longer time in the high-risk group than that in the normal-risk group [(30.6±0.11) min vs (19.1±0.09) min, P<0.05]. The number of needle punctures, percent of successful cannulation, and the frequency of immediate complications were similar in the high- and normal-risk groups. It was more likely to form catheter thrombosis during long-term use in the high-risk group (4/20, 20%) which might cause poor blood flow.@*CONCLUSION@#Permanent IJV cannulation under real-time ultrasound guidance is very safe with high success rates. Nephrologists can use this technique with ease and with minimal complications in both normal- and high-risk patients.

Indoleamine-pyrrole 2,3-dioxygenase might be a prognostic biomarker for patients with renal cell carcinoma / 中南大学学报(医学版)

Objective:To explore the role of indoleamine-pyrrole 2,3-dioxygenase (IDO),an immunomodulatory enzyme,in renal cell carcinoma (RCC).Methods:A total of 40 patients diagnosed as RCC in the Second Xiangya Hospital were included in this study.All patients received nephrectomy.The histopathological features of samples were assessed semi-quantitatively.IDO mRNA level in RCC and non-RCC renal tissues was determined by real-time quantitative PCR (RT-qPCR).And the expression of IDO protein in endothelial cells was examined by immunohistochemistry; a Kaplan-Meier survival curves was calculated on the basis of IDO mRNA level.Results:Level of IDO mRNA in RCC samples was significantly higher than that in tumor-free samples with P＜0.001.Patients with high IDO expression had an significantly longer survival time than those with low IDO expression (P=0.01).There was a statistically significant inverse correlation between IDO and proliferation marker Ki67.Patients with high IDO level were of low Ki67 level,and vice versa (P＜0.01).Conclusion:IDO might be a prognostic biomarker for patients with RCC.

New pathologic classification of diabetic nephropathy (retrospective study of 37 cases) / 中南大学学报(医学版)

OBJECTIVE@#To investigate the new pathological classification of diabetic nephropathy (DN) published by Research Committee of the Renal Pathology Society in 2010.@*METHODS@#Renal biopsy specimens were obtained from 37 patients with type 2 diabetes mellitus with micro-albuminuria (MAU) or clinical albuminuria (CAU). These samples were classified according to new pathological classification for DN and new standard scores for interstitial vascular injury.@*RESULTS@#Before the classification, DN was seen in 26 palients. After re-analysis according to the new pathological classification, the patients diagnosed with DN increased to 32. In these 32 DN patients, 1 was classified as type I, 3 as type IIa, 2 as type IIb, 23 as type III and 3 as type IV; 12 patients had mild interstitial injury, 15 had midrange interstitial injury, while 5 had severe interstitial injury.@*CONCLUSION@#The new pathological classification of DN can increase the diagnosis rate and attract more attention to tubular and interstitial damage in DN, contributing to the early diagnosis and treatment of DN.

Role of galectin-1 on epithelial-to-mesenchymal transition induced by high glucose peritoneal dialysate in human peritoneal mesothelial cells / 中南大学学报(医学版)

OBJECTIVE@#To investigate the expression of galectin-1 with the stimulation of peritoneal dialysis solution (PDS) and its role in the epithelial-to-mesenchymal transition (EMT) in human peritoneal mesothelial cells (HPMCs).@*METHODS@#HPMCs were stimulated with PDS containing different concentrations of high glucose (1.5%, 2.5%, and 4.25%). After 24 h, mRNA and protein expressions of galectin-1,vimentin, and zo-1 were analyzed with real-time PCR and Western blot, respectively. Liposome transfected siRNA technique was used to knock down the expression of galectin-1 and the effect of galectin-1 siRNA on the EMT of HPMCs was also observed under 4.25% PDS condition.@*RESULTS@#mRNA expression of galectin-1 in HPMCs increased in PDS groups, especially in group with 4.25% PDS (P<0.05). Protein expression of galectin-1 in HPMCs significantly increased in PDS groups with a dose dependent manner (P<0.05).Expression of vimentin in HPMCs significantly increased in PDS groups, especially in groups of 2.5% PDS and 4.25% PDS (P<0.05), but zo-1 expression markedly decreased (P<0.05). The expression of galectin-1 correlated positively with vimentin (P<0.05) but negatively with zo-1 (P<0.05). Expression of vimentin in groups of 4.25% PDS was markedly inhibited (P<0.05) by galectin-1 siRNA, whereas zo-1 expression was significantly increased (P<0.05).@*CONCLUSION@#Galectin-1 can mediate high glucose PDS-induced EMT in HPMCs and may be a new target for the prevention and treatment of peritoneal fibrosis.

Expression of B1a cells and IgA1 positive cells in tonsil of IgA nephropathy patients and analysis of associated clinicopathological factors / 中华肾脏病杂志

Objective To examine the expression of IgA1 and B1a positive cells in palatine tonsils of IgA nephropathy (IgAN) patients, and to analyze the association between B1a cells and clinicopathological changes. Methods Eight patients diagnosed as IgAN by renal biopsy and 8 chronic tonsillitis patients without nephritis as control were enrolled in the study.Immunofluorescence and laser scanning confocal microscope (LSCM) were applied to observe the localization and quantitative calculation of Bla and IgA1 positive cells. Statistic analysis of the association of B1a cells with proteinuria and pathological Lee's grading was performed. Results Bla cells were mainly localized in germinal center of tonsil, and IgA1 positive cells were mainly localized in subepithelium of tonsil. Compared to control group, the percent of B1a cells and IgA1 positive cells was significantly higher in IgAN (P＜0.01). There was a positive correlation between Bla cells and IgA1 cells (P＜0.05). In IgAN, the percent of B1a cells in patients with hematuria and proteinuria was obviously higher than that of patients with hematuria only (P＜0.05). The number of Bla cells in IgAN patients with≥Lee's grade Ⅲ was significantly higher than that of those ＜ grade Ⅲ (P＜0.05). Conclusions IgA1 may be secreted by Bla cells in the tonsil of IgAN patients. The number of B1a cells is correlated with exacerbation of proteinuria and pathological severity, which may play an important role in pathogenesis of IgAN.

Effects of high levels of glucose on the expression of adiponectin receptors in human kidney proximal tubular cells / 中国医师杂志

Objective To investigate the effect of in vitro high glucose stimulation on the expression of adiponectin receptor (adipoR) in human kidney proximal tubular cells.Methods The HK-2 cells were cultured in the low glucose DMEM culture medium containing 10% fetal bovine serum until the cells were adherent and 80% confluence. After cultured in the serum-free DMEM for 24 hours, these cells were stimulated with glucose-containing 1mg/ml, 2mg/ml, 4mg / ml, 6mg/ml, 8mg/ml serum-free DMEM for 48 hours. Then RT-PCR and western blot were used to analyze adipoR ( R1, R2) expression levels. The HK-2 cells were cultured respectively in high glucose (4mg/ml) , low glucose (1mg/ml) DMEM culture medium containing 10% fetal bovine serum to cultivate 0h, 12h, 24h, 48h, 72h, 96h, then RT - PCR was applied to analyze adipoR (R1, R2) mRNA expression levels semi-quantitatively. Results Two kinds of adiponectin receptor gene were both expressed in HK-2 cells, and the quantity of gene expression of adipoR1 (0. 63 ±0. 12) was 3. 9 times to adipoR2 (0. 16 ±0.03) , the difference was statistically significant ( P<0. 01). The different concentrations of glucose and different time of high glucose on HK-2 cells had no significant effect ( P>0. 05 ) on adipoR gene expression. Expression of adipoR 1 protein in HK-2 cells was detected by western blot, and it was not affected by glucose concentration ( P>0. 05).Conclusion adi-poR1 and adipoR2 gene were both expressed in HK-2 cells, and the adipoR1 was the major one, which suggested that adipoR1 played a more significant role in kidney disease. The expression of adipoRl/R2 of HK-2 cells was not affected by high glucose concentration.

Role of mitochondrial respiratory chain in hyperpermeability of human peritoneal mesothelial cells induced by high glucose peritoneal dialysate / 中华肾脏病杂志

Objective To investigate the role of mitochondrial respiratory chain in the hyperpermeability of human peritoneal mesothelial cells (HPMCs) induced by high glucose peritoneal glucose PDS was also added. Transmesothelial electrical resistance (TER) measurement was examined for detection of permeability damage in HPMCs. Immunostaining and Western blotting analysis were used to detect claudin-1 expression. Mitochondrial superoxide (MitoSOX) Red staining and respiratory chain complexes activities were determined for detection of mitochondrial reactive oxygen species (ROS) production and mitochondrial complexes activities. Results TER was decreased in a time- and concentration-dependent manner after culture with high glucose PDS for was also down-regulated significantly by high glucose PDS (P＜0.01). Complex Ⅲ activity was inhibited (10.8% of control, P＜0.01) accompanied with increased mitochondrial ROS generation.These changes were partially prevented by glutathione. Conclusion Mitochondrial respiratory complex Ⅲ pathway has crucial importance in maintaining TER of HPMCs, which may reveal a valuable target for novel therapies to fight hyperpermeability of peritoneum during the prolonged PD treatment.

Stages of 3,547 patients with chronic kidney disease and relevant factor analysis / 中南大学学报(医学版)

OBJECTIVE@#To investigate the prevalence and distribution of chronic kidney disease(CKD) in Second Xiangya Hospital of Central South University.@*METHODS@#We retrospectively analyzed the medical records of 3547 CKD patients (>or=14 years old) admitted to Second Xiangya Hospital for the 1st time from January 2003 to December 2008. Glomerular filtration rate (GFR) was estimated by using the MDRD equation abbreviated [eGFR=186.3*SCr-1.154*age-0.203*0.742 (for women) mL/min.1.73 m2]. Data of patients' gender, age, admission number, etiologic and functional diagnose, course of disease, blood pressure, hemoglobin, urine, renal function, blood albumin, and kidney ultra-B were collected.@*RESULTS@#(1) The most common causes for CKD were primary glomerulonephritis(55.20%), hypertension (14.55%) and diabetes (11.78%). The composition of the causes during this 6 years was not significantly different (P>0.05). The proportion of chronic glomerulonephritis declined while that of hypertensive nephropathy and diabetic nephropathy increased as time passed. (2) Patients were respectively distributed from stage 3 to stage 5 at the following percentages: 11.59% (stage 3), 23.03% (stage 4), and 65.38% (stage 5). Middle-aged and young patients (41 years60 years) accounted for 36.88% of the CKD patients. Chronic glomerulonephritis was often seen at 21-40 and 41-60 year olds. Patients with lupus nephritis were younger, 48.35% of whom were 21-40 years old. On the contrary, patients with hypertensive nephropathy and diabetic nephropathy were much older. They were mainly seen in older than 40 year groups. (3) Obstructive nephropathy and gout nephropathy occurred mainly in men while it seemed much easier for women to suffer from lupus nephritis. (4) The majority of CKD patients was accompanied by anemia (94.28%) and hypertension (56.91%), and the incidence and degree of anemia and hypertension increased as CKD developed (P<0.005).@*CONCLUSION@#The top 3 causes of CKD are chronic glomerulonephritis, hypertensive nephropathy, and diabetic nephropathy. The proportion of hypertensive nephropathy and diabetic nephropathy is on the rise. Elderly people are the high risk group for CKD.

SARA expression in high glucose induced epithelium to mesenchymal transition of proximal tubule cells / 中南大学学报(医学版)

OBJECTIVE@#To construct the cell model of epithelium to mesenchymal transition of proximal tubule cells induced by high glucose and to determine the expression of Smad anchor for receptor activation (SARA).@*METHODS@#Protein expression of vimentin, Zona occludens-1(ZO-1), and SARA was determined by Western blot, and their mRNA expressions were detected by Real-time PCR.@*RESULTS@#After stimulation by 30 mmol/L D-glucose, the protein and mRNA expression levels of vimentin in HK-2 cells increased in a time-dependent manner while the expression of ZO-1 was reduced significantly, especially at 48 h. Meanwhile, SARA was also decreased in a time-dependent manner.@*CONCLUSION@#High glucose can induce renal epithelium to mesenchymal transition, and SARA may be involved in this process as a protector.

Effect of glucose peritoneal dialysates on the transmesothelial electrical resistance and cellular migration of monolayer human peritoneal mesothelial cell / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of different concentrations of glucose peritoneal dialysates (PDS) on monolayer transmesothelial electrical resistance (TER) and migration ability of cultured human peritoneal mesothelial cells (HPMCs) to clarify the cause of peritoneal hyperpermeability state and ultrafiltration failure during prolonged peritoneal dialysis.@*METHODS@#HPMCs were cultured in a 1:1 mixture of DMEM and PDS containing 1.5%, 2.5%, and 4.25% glucose. Methyl thiazolyl tetrazolium (MTT) assay and TER were measured to determine the effect of glucose PDS on the proliferation and permeability of human peritoneal mesothelial monolayers, respectively. Wound-healing assay was used to confirm whether glucose could do harm to the migration of cells.@*RESULTS@#Proliferation of HPMCs was significantly suppressed by different glucose concentrations at 24 hours. TER decreased in a time- and concentration-dependent manner after culture with different concentrations of glucose PDS. Cells lost migration in the presence of high glucose after 24 hours, and most cells lost their normal morphology and became detached from plates after 48 hours of wounding.@*CONCLUSION@#High glucose in PDS can cause peritoneal damage by suppressing cell proliferation, inducing increase in paracellular permeability of HPMCs and inhibiting cell migration after damage, which may be responsible for peritoneal hyperpermeability and the development of ultrafiltration failure.

Basic experimental and clinical research on peritoneal dialysis in the past 16 years / 中南大学学报(医学版)

To summarized the experiences from our basic experimental and clinical research on peritoneal dialysis. In the past 16 years, peritoneal fibrosis rat models and rabbit models of peritonitis were first established successfully in our laboratory in China. Peritoneal mesothelial cells were also separated and identificated. Besides, we assessed the biocompatibility of peritoneal dialysis fluid and analyzed the molecular mechanism of peritoneal mesothelial cell injury. We demonstrated the key role of transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF) and peroxisome proliferative activated receptor-gamma (PPAR-gamma) in the pathogenesis of peritoneal fibrosis, as well as their regulation of molecular mechanism. Furthermore, we transfected the plasmids encoding TGF-beta1-shRNA or pCTGF-shRNA into peritoneal cells and tissues by nanocarrier technologies. In clinical research, the positioning of peritoneal dialysis catheters, peritoneal dialysis treatment modalities and the prevention and treatment of its complications were studied. The characteristics and mechanism of solute transport in peritoneal dialysis was also explored.

Risk factors for acute kidney injury following 5100 cardiac surgeries with extracorporeal circulation / 中南大学学报(医学版)

OBJECTIVE@#To determine the incidence and risk factors associated with acute kidney injury (AKI) in patients after cardiac surgery with extracorporeal circulation.@*METHODS@#A retrospective case control study was done in patients who underwent cardiac surgery from 2003 to 2007 in Second Xiangya Hospital, with 340 patients in an AKI group and the other 4 760 patients without AKI as a control group. All variables were analyzed by univariate analysis, Mann-Whitney U test and logistic regression.@*RESULTS@#AKI occurred in the 340 patients (6.7% incidence). Univariate analysis revealed that age, preoperative serum creatinine, preoperative ejection fraction (EF), preoperative beta2-microglobulin, preoperative blood albumin, preoperative blood uric acid, intraoperative cardiopulmonary bypass time, intraoperative aortic cross-clamp time, and dosage of mannitol were significantly related to AKI following cardiac surgery with extracorporeal circulation. Logistic multivariate regression analysis showed that preoperative serum creatinine (P<0.001), preoperative ejection fraction (EF) (P<0.001), preoperative beta2-microglobulin (P=0.002), preoperative blood uric acid (P=0.015), intraoperative cardiopulmonary bypass time (P<0.001), and intraoperative aortic cross-clamp time (P<0.001) were independent risk factors for AKI.@*CONCLUSION@#The incidence of AKI after cardiac surgery with extracorporeal circulation is closely related with a variety of perioperative risk factors. Our data suggest that patients planning to accept cardiac surgery with extracorporeal circulation should be more comprehensively assessed and monitored, thereby preventing the occurrence of AKI.

Bacterial biofilms on PVC tubing's inner surface of hemodialysis water treatment system / 中南大学学报(医学版)

Objective To determine the morphology, bacteria and endotoxin content of bio-films on the inner surface of PVC tubes in hemodialysis water treatment system. Methods We dissolved biofilms of segments before and after reverse osmosis machine for bacterial count and identification. We studied biofilm structure of segments before and after reverse osmosis machine with eyes and scanning electron microscope. Biofilms of all 7 segments were dissolved for qualitative and quantitative assay of endotoxin. Results The inner surface of segment before reverse osmosis machine was homogeneously distributed with activated carbon powder deposition. The segment after reverse osmosis machine was normal. With scanning electron microscope, biofilm with successive surface and sandwich was found on the inner surface of segment before reverse osmosis machine, formed by clustering bacillus, activated carbon powder and some coccus. Bacteria of the same shape and length were found on segment after reverse osmosis machine, but fewer and looser. Bacterial culture and identification showed the former was mostly gram-negative bacillus, the latter was only a few micrococcus. Endotox-in of biofilm was between 2. 0 EU/mL and 4. 0 EU/mL. Quantitative assay showed: segment after softener (2.821 ±0. 807) EU/mL; segment after active charcoal canister(3. 635 ±0. 427) EU/ mL; segment before reverse osmosis machine (3.687 ±0.271) EU/mL; segment after reverse osmosis machine (2. 041 ±0. 295) EU/mL; exit of power pump (1. 983 ±0. 390) EU/mL; the 1st dead space (2. 373 ± 0. 535) EU/mL; and the 2nd dead space (2. 858 ± 0. 690) EU/mL. Conclusion Biofilms are found on the inner surface of segment before and after reverse osmosis machine . Endotoxin level from high to low is as follows: segment before reverse osmosis machine, segment after active charcoal canister, the 2 nd dead space, segment after softener, the 1 st dead space, segment after reverse osmosis machine, exit of power pump. The character of the bacteria and endotoxin of the biofilm can help us find better ways to control them.

Mast cell infiltration is involved in renal interstitial fibrosis of rat models with protein overload nephropathy / 中华肾脏病杂志

Objective To investigate the correlation of infiltration of mast cells in kidney with renal interstitial fibrosis, expression of TGF-β1 and stem eel] factor (SCF) in rat models withprotein-overload nephropathy. Methods Sixty uninephrectomized SD rats were randomly divided into model group [intraperitoneal injections of bovine serum albumin (BSA)] and control group (intraperitoneal injections of equal volume of saline). Ten rats from both groups were sacrificed respectively at week 3, 7 and 11 after injection. 24 h urinary protein and serum biochemistry of these SD rats at the time of sacrifice were measured. The intensity of mast cell infiltration was examined by toluidine blue (TB) staining and immunohistochemistry using a monoclonal anti-MC chymase antibody. The expression of TGF-β1 and SCF was detected byimmunohistochemistry, using a monoclonal mouse anti-rat TGF-β1 antibody and a polyclonal rabbstanti-rat SCF antibody. Results Severe proteinuria was induced in the rats by BSA injectionpeaked at week 7 [(199.1±98.4) mg/d] after the BSA injection and gradually decreased until week11 [(133.7±67.8) mg/d]. Renal injury was accompanied with chymase-postitive and TB-postitive mast cell infiltration, in close proximity to areas of interstitial fibrosis. With aggravation oflesions degree, the number of mast cells increased,the difference between the modal rats and control rats was significant (P<0.05). Immunostainahle expression of SCIF and TGF-β1 was detected in tubular as well as interstitial cells, and increased with the BSA injection. The difference between the model rats and control rats was significant (P<0.05). Mast cells were positively correlated with interstitial fibrosis (r=0.772, P<0.01), expression of TGF-β1 (r=0.521, P<0.01) and SCF(r=0.916,P<0.01). Conclusions Increased infiltration of mast cells is involved in interstitial fibrosis of rats with protein-overload nephropathy. Proteinuria may attract mast cells to kidney by chemot actions of SCF,and mast cells may contribute to the development of renal fibrosis by secreting chymase and increasing expression of TGF-β1.

PRS-CTGF-siRNA attenuates matrix production and the expression of vascular endothelial growth factors induced by transforming growth factor β1 in cultured human peritoneal mesothelial cells / 中华肾脏病杂志

Objective To investigate the effects of connective tissue growth factor (CTGF) siRNA delivered by pRetro-Super (PRS) retrovirus vector on extracellular matrix and VEGF expression in human peritoneal mesothelial cells (HPMC). Methods Four pairs of oligonucleotides including 64 bp DNA were designed and synthesized in vitro according to siRNA target sequence and PRS retrovirus desire.PRS-CTGF-siRNA1-4 recombinant retrovirus vectors were constructed.The recombinant retrovirus vectors containing CTGF-siRNA were transferred into PT67 packaging cell lines with lipefectamine 2000,then infected HPMC.mRNA expression was determined by semi-quantitative RT-PCR and protein expression was determined by Western blot.Results Both mRNA and protein expressions of CTGF,FN,Col I,laminin (LN) and VEGF were significantly increased in HPMC with 5 μg/L TGF-β1 stimulation (P＜0.01,respectively).CTGF,FN,Col I,LN mRNA and protein and VEGF mRNA expression stimulated by TGF-β1 were significantly decreased in HPMC infected with PRS-CTGF-siRNA1～4 retrovirus vectors (P＜0.01,respectively).The inhibitory rates on CTGF were 69.3%,22.2%,27.4% and 38.8%,respectively (P＜0.01).At the same time,there was also a significant reduction of VEGF protein expression in HPMC infected with PRS-CTGF-siRNA1 vector (P＜0.01).There was no significant difference in HPMC infected with PRS void vector. Conclusion CTGF siRNA delivered by PRS retrovirus vector can effectively inhibit the enhancement of extracellular matrix and VEGF expression stimulated by TGF-β1 in HPMC.

Effect of dialysate composition on proliferation of human peritoneal mesothelial cells / 中国医师杂志

Objective To make sure the effect of dialysate composition on HPMCs.Methods Cell strains were subculturing.There are six groups in this experiment: Group 1(control);group 2(4.25% Glucose);group 3(1.75mmol/L Ca~(2+));group 4(1.25 mmol/L Ca~(2+));group 5(4.25% Glucose+1.75mmol/L Ca~(2+));group 6(4.25% Glucose+1.25mmol/L Ca~(2+)).The capacity of proliferation of HPMCs was assessed by MTT assay. Results Proliferation of HPMCs was inhibited in 4.25% glucose group in time dependence.Ca~(2+) induce proliferation of HPMCs,however,no effect on proliferation of HPMCs exists in different Ca~(2+) group.Conclusion High glucose can inhibit cell proliferation;Ca~(2+)(1.75mmol/L,1.25mmol/L) can promote the proliferation and 1.25mmol/L is better.