AIM: To investigate the expression of the nephrin in podocyte of the diabetic nephropathy(DN) rats and the mechanism of irbesartan-induced renal protection.METHODS: The DN model was established by a single injection of streptozotocin(STZ),and DN rats were randomly divided into 2 groups: model group and irbesartan treatment group.In addition,the normal rats served as a normal control group. All the rats were received daily gavage respectively for 8 weeks. The urinary protein quality in 24 hours,body weight(BW),kidney weight (KW),KW/BW,glucemia,urea nitrogen,creatinine,total cholesterol, triacylglycerol were detected with correlative methods and the pathological changes of kidney were also detected with optic microscope and transmission electron microscope.The expression of nephrin in podocyte were detected by immunohistochemistry. RESULTS: In DN rats, irbesartan reduced the urinary protein quality in 24 hours (P

BACKGROUND: Diabetic nephropathy is one of the most serious vascular complications of diabetes mellitus. Compound preparation of huangqi and dahuang, a traditional Chinese medicine, has been used to preventing or treating diabetic nephropathy for several years, and has a certain protective effect on the kidney of diabetes mellitus patients. But its exact mechanism remains unknown and needs to be studied more.OBJECTIVE: To investigate the effect of compound preparation shenkang wan on the proliferation and secretion of extracellular matrix in cultured rat mesangial cells induced by high glucose.DESIGN: Randomized and controlled study.SETTING: Center of Integrated Traditional and Western Nephrology of Zhujiang Hospital and Medicine Department of Nanfang Hospital, Southern Medical University.MATERIALS: The serum pharmacological experiment was performed in Animal Experimental Center of Southern Medical University in A pril 2005.The cell culture experiment was conducted in Cell culture room of Southern Medical University from April 2005 to July 2005. Totally 16 normal Wistar male rats, weighted varied from 190 g to 220 g, were used in the study.METHODS: Sixteen normal Wistar male rats were randomly divided into 4 groups: normal serum group, capoten group, shenkang wan group (high dose and low dose); shenkang wan was mainly constituted of huangqi,dahuang, leech, gordon guryale seed and corn stigma and made in Pharmacy Department of Zhujiang Hospital of Nanfang Medical University, agent number: 20031214). ① The rats in capoten group and high and low dose shenkang wan group were given the corresponding drugs respectively according to 5 mg/kg, 2.4 g/kg, 1.2 g/kg weight. The rats in normal serum group were given the same volume water. After treated 7 days, all rats were hocused and separated medication serum. ② Mesangial cell was cultured in vitro with different concentrations of glucose (10, 20, 30 and 40 mmol/L).The proliferation of mesangial cell was observed with the methyl-thiazoltelrazolium colorimetric assay at 24, 48, 72 hours and 96 hours. ③ Then the cultured mesangial cells were divided into six subgroups :Low glucose control group (10 mmol/L glucose), high glucose group (30 mmol/L glucose);normal serum group (30 mmol/L glucose); capoten group (30 mmol/L glucose); shenkang wan group (high dose and low dose, 30 mmol/L glucose).After cultured 72 hours, the proliferation of mesangial cell was detected with the methyl-thiazol-telrazolium colorimetric assay, the secretion and mRNA gene expression of fibronetin levels in mesangial cell were respectively detected by enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) method.MAIN OUTCOME MEASURES: ①Proliferation of mesangial cell induced by different concentrations glucose. ② Proliferation and secretion and mRNA gene expression of fibronectin in every group.RUSULTS: ① Effect of different concentrations glucose on the prolifera-tion of mesangial cell: Compared with low concentrations glucose(10 mmol/L), 20 mmol/L glucose could accelerate the proliferation ofmesangial cell during 96 hours experiment period, but only had a statisti-cally significant difference at 72 and 96 hours (P ＜ 0.05). 30 mmol/L glu-cose could significantly accelerate the proliferation of mesangial cell thanthat of 10 mmol/L glucose from 24 hours to 96 hours (P ＜ 0.05 or P ＜ 0.01),and this effect was increasing with time in 72 hours and reduced after 72hours. 40 mmol/L glucose could significantly increase the proliferation ofmesangial cell than of low concentrations glucose in 48 hours (P ＜ 0.05),and this effect was reduced after 48 hours and even conversed to restraineffect. ② Effect of different medication serum on the proliferation ofmesangial cell: The optical density value in high glucose group is obviouslyhigher than that of low glucose control group (P ＜ 0.01). Compared withhigh glucose group, the optical density value in capoten, shenkang wangroup (high dose and low dose) was decreased markedly (P ＜ 0.01 or P＜ 0.05). While the optical density value in normal serum group was showedno difference with the high glucose group (P ＞ 0.05). ③ Effect of differentmedication serum on secretion of fibronectin in mesangial cell: Content offibronectin in high glucose group was increased more markedly than that oflow glucose group (P ＜ 0.01). Compared with high glucose group, contentof fibronectin in capoten and shenkang wan group (high dose and low dose)was showed a significantly decrease (P ＜ 0.01 or P ＜ 0.05), while contentof fibronectin in normal serum group was showed no difference with thehigh glucose group (P ＞ 0.05). ④ Effect of different medication serum onexpression of fibronectin mRNA in mesangial cell: The optical density val-ue of fibronectin strip in high glucose group was brighter than that in lowglucose group and the ratio of it and β-actin were increased markedly too(P ＜ 0.01). Compared with high glucose group, the optical density value offibronectin strip in capoten and shenkang wan group (high dose and lowdose) was showed a significantly decrease and the ratio of it and β-actinwas reduced distinctly too (P ＜ 0.01), while the ratio of it and β-actin innormal serum group was showed no difference (P ＞ 0.05).CONCLUSION: High glucose could accelerate proliferation, increase thesecretion and mRNA gene expression of fibronectin in mesangial cell,while shenkang wan could inhibit proliferation and secretion of the extra-cellular matrix in mesangial cell induced by high glucose.

AIM: To investigate the clinical effect of Maiditong for injection(Herba Erigerontis) on coagulative and fibrionlytic system in diabetic nephropathy(DN). METHODS: 59 patients definitely diagnosed as diabetes type 2 DN were randomly divided into treatment and control groups.The treatment group were treated with Maiditong for injection,and the control group were treated with of Compound Danshen Injection.The 24 h proteinuria and vWF:Ag、Fbg、PF_(1+2)、FPA、D-dimer、AT-Ⅲ:A、tPA、PAI-1、PIC、PLG were tested before and after the therapies. RESULTS: After one therapy course(2 weeks),in the treatment group the 24 h proteinuria decreased obviously(P

BACKGROUND:There is lack of effective diagnostic methods for early knee osteoarthritis. Proteomics refers to the large-scale experimental analysis of proteomes from the overal level of intracel ular protein compositioon, expression and modification, based on which, we can understand protein-protein interaction and relationship, thereby revealing protein functions and cel activity patterns. OBJECTIVE:To apply the isobaric tags for relative and absolute quantitation (iTRAQ) to identify proteins differential y in serums of osteoarthritis Kel gren-Lawrence classification at each stage, and to find out potential molecular markers at each stage of osteoarthritis. METHODS:Sixty patients with knee osteoarthritis were enrol ed, and according to Kel gren-Lawrence (K-L) grading, they were subdivided into K-L 0, II and IV groups. There were 10 males and 10 females randomly selected in each group. Proteins differential y regulated in serums were identified by the stable isotopes 116, 117, 118 of the iTRAQ labeled, reversed-phase column separation, mass spectrometry and Swissport database search, after the serums were subjected to high abundance proteins depletion. Final y, we analyzed the proteins identified by using bioinformatics software. RESULTS AND CONCLUSION:Total y 169 proteins were identified through iTRAQ peptides experiments of different sample tags, Q-star mass spectrum identification and MASCOT library search. 153 proteins were identified as K-L 0 to K-L IV, 153 proteins were identified as K-L 0 to K-L II, and 145 proteins were identified as K-L II to K-L IV. iTRAQ technology may help to identify novel serum markers for early diagnosis of knee osteoarthritis, indicating iTRAQ technology for proteomics serum biomarkers of osteoarthritis study has good application prospect.

Objective To investigate the incidence and influencing factors of aldosterone breakthrough during therapy with angiotensin Ⅱ receptor blockers (ARB) alone,or combined with angiotensin-converting enzyme inhibitors (ACEI) in Chinese patients with non-diabetic nephropathy.Methods A total of 144 patients with non-diabetic nephropathy were treated with ARB or combination therapy of ACEI and ARB for a mean follow-up period of 12 months.Aldosterone breakthrough was determined according to the change of plasma aldosterone concentration before and after treatment during 6-month and 12-month ACEI/ARB treatment.Results In 6 months,aldosterone breakthrough occurred in 21 patients,corresponding to 14.58％,while in 12 months,occurred in 39 patients,corresponding to 27.08％.Although the overall urinary protein excretion (UPE) decreased after treatment in both groups (P＜0.05),non-breakthrough group had a more remarkable reduction in UPE (P＜0.05).Univariate Logistic regression demonstrated that risk factors of aldosterone breakthrough included pre-treatment values of UPE (OR=3.643,P=0.073) and eGFR (OR=0.980,P=0.025).Multivariate Logistic model revealed pre-treatment values of eGFR was positively associated with aldosterone breakthrough (OR=0.980,P=0.025).Conclusions The incidence of the aldosterone breakthrough increases with duration of treatment.The patients with aldosterone breathrough have higher level of UPE,and enhanced decline in eGFR.Pretreatment value of eGFR is independent risk factor of aldosterone breakthrough.

Aim To investigate the effect of gelsemium alkaloids on chloride channels and cell volume in he-patic carcinoma cells. Methods The time-lapse live cell imaging and whole-cell patch clamp techniques were used respectively to detect the volume changes and currents induced by gelsemium alkaloids in HepG2 cells. Results It was found that the cell volume was decreased by (12. 48 ± 2. 2) % (P<0. 01) when ex-posed to gelsemium alkaloids for 50 min and this phe-nomenon could be inhibited by the chloride channel blocker tamoxifen. It was shown by whole-cell patch clamping that a chloride current could be evoked by extracellular application of gelsemium alkaloids ( 2μmol·L-1 ) . The current was outward-rectified with-out obvious voltage- and time-dependent inactivation. The reversal potential of the current was ( -3. 21 ± 0. 67) mV ,which was close to the equilibrium poten-tial of chloride. The extracellular application of the chloride blockers, tamoxifen and 5-notro-2-(3-phenyl-propylamino)benzoic acid (NPPB), and 47% hyper-tonic solution inhibited the current significantly ( P <0. 01 ) . Conclusion Gelsemium alkaloids could acti-vate chloride channels and induce a volume decrease ( named apoptotic volume decrease, AVD) , and these effect could be inhibited by chloride channel blockers. The results suggest that the chloride channel can be one of the targets of gelsemium alkaloids in their anti-cancer action.

The Cathepsin L-like cysteine proteinase genes (cpls) are multifunction genes related to the parasitic abilities of plant parasitic nematodes. A new cathepsin L-like cysteine proteinase gene (Dd-cpl-1) (GenBank Accession GQ 180107) was cloned from Ditylenchus destructor by RT-PCR and RACE. The cDNA sequence consisted of a 1 131 bp open reading frame (ORF) encoding 376 amino acid residues that were franked by a 29 bp 5'-untranslated region (UTR) and a 159 bp 3'-UTR. Genomic sequence analysis showed that Dd-cpl-1 contained 7 introns, obeyed the GT/AG rule in the splice-site junctions. Homology analysis showed that the identity was 77% between Dd-cpl-1 deduced protein Dd-CPL-1 and cathepsin L-like cysteine proteinase of Bursaphelenchus xylophilus. Multi-sequence alignment indicated that there were the catalytic triad (Cys183, His322 and Asn343) and two motifs ERFNIN motif and GNFD motif in deduced protein Dd-CPL-1. Cysteine proteinases phylogenetic analysis showed that Dd-cpl-1 belonged to the sub-clade of cathepsin L-like cysteine proteinases.
Amino Acid Sequence ; Animals ; Cathepsin L ; genetics ; Cloning, Molecular ; Cysteine Proteases ; genetics ; Genes, Helminth ; genetics ; Molecular Sequence Data ; Nematoda ; enzymology ; genetics ; Phylogeny ; Sequence Alignment ; Sequence Analysis, Protein ; Sequence Homology, Amino Acid ; Solanum tuberosum ; parasitology

Aim To clarify the effect of Borneol on the chloride channels and cell volume in poorly differentia-ted nasopharyngeal carcinoma CNE-2Z cells.Methods The technique of whole-cell patch clamp was used to detect the chloride currents and analyze the character-istics of the currents in CNE-2Z cells.The volume changes caused by Borneol were measured by the meth-od of time-lapse live cell imaging.Results The chlo-ride currents were induced by extracellular application of Borneol (20 μmol·L -1 )isotonic condition.The currents showed a characteristic of outward rectification and did not show voltage-dependent or time-dependent inactivation.The reversal potential of the currents was close to the CI-equilibrium potential. The currents were inhibited by the chloride channel blocker tamox-ifen.The currents were also inhibited by 47% hyper-tonic solution.Borneol decreased the cell volume by 9.4% in 30 min.Tamoxifen completely inhibited the Borneol-induced cell volume decrease.Conclusion Borneol can activate volume-sensitive chloride channels and induce volume decrease in CNE-2Z cells.Chloride channels play a pivotal role in the process of volume decrease caused by Borneol.

Objective:To estimate in-hospital mortality after knee replacement (KR) and to assess its trend and risk factors in China.Methods:We included patients undergoing KR in the Hospital Quality Monitoring System in China (2013-2019) to estimate in-hospital mortality after KR and assessed relation of patient's and hospital's characteristics (year of surgery, age, gender, marital status, primary indication, Charlson comorbidity index, geographic location, hospital type, hospital volume of KR, and surgery type) to in-hospital mortality using multivariable Poisson regression.Results:The annual amount of KR has increased from 20 307 in 2013 to 35 757 in 2019, and has maintained an upward trend for 7 years. The mean age of patients having KR increased from 64.9 years in 2013 to 66.6 years in 2019. Of the total 218 923 KRs, 63 deaths (0.29‰) occurred within 30 days before discharging. Older age was associated with higher in-hospital mortality ( P for trend <0.001). Male gender had higher incidence of in-hospital mortality compared with female [relative risk (RR), 2.5; 95% CI: 1.5, 4.1]. Single marital status was associated with higher, albeit non-statistically significant, in-hospital mortality than married patients (RR, 2.1; 95% CI: 0.9, 4.6). Higher Charlson comorbidity index was associated with increased risk of in-hospital mortality ( P for trend <0.001). Risk of in-hospital mortality decreased with more hospital-year knee replacement surgeries ( P for trend <0.001). In-hospital mortality varied by geographic regions, with the lowest mortality in East region (0.16‰), followed by South-West (0.31‰), South-Central (0.31‰), North region (0.33‰), North-West (0.54‰) and North-East (0.59‰). Conclusion:In-hospital mortality after KR in China was relatively low. Older age, male gender, higher Charlson comorbidity index and lower hospital-year knee replacement surgeries were risk factors for in-hospital mortality. The mortality varied greatly according to the geographic location of hospital.

Diabetic ketoacidosis (DKA), a serious acute complication of diabetes mellitus, mainly manifests as hyperglycemia, ketosis, and acidosis. It is a metabolic syndrome resulting from insulin deficiency and increased insulin-antagonistic hormone levels. While type 2 diabetes mellitus complicated by DKA is relatively uncommon, secondary pneumomediastinum in DKA is extremely rare. Following alveolar rupture, air can travel through various routes to reach the hilum, causing anterior, middle, or posterior pneumomediastinum or even leading to intracranial epidural pneumatosis. The diagnosis of pneumomediastinum is mainly dependent on chest computed tomography findings. After the successful treatment of DKA, pneumomediastinum usually resolves spontaneously within 5-10 days with a good prognosis. One DKA patient admitted to Dege County People's Hospital developed Kussmaul respirations, followed by an increase in intra-alveolar pressure, an elevation in intra and extra-alveolar pressure difference, and protein decomposition in the alveolus wall, which promoted alveolar rupture and induced mediastinal emphysema. After rapid fluid replacement, blood glucose control with insulin, and maintenance of acid-base balance (correction DKA), the mediastinal emphysema was spontaneously absorbed. Through the analysis of the clinical data of this case, the purpose is to improve the clinicians' internal understanding of the relationship between mediastinal emphysema and DKA, avoid over-examination and over-treatment, and provide strategies for correct diagnosis and treatment.