Effects of endothelial nitric oxide synthase gene on end stage renal disease progression in autosomal dominant polycystic kidney disease.

AIM: To investigate whether endothelial nitric oxide synthase (eNOS) gene associate with the progression of autosomal dominant polycystic kidney disease (ADPKD). METHODS: Databases of EMBASE, Pubmed, ISI, Ovid Database, Cochrane library and China National Knowledge Infrastructure were all searched. Associated studies about eNOS polymorphisms and ADPKD were analyzed by meta-analysis. RESULTS: A total of 11 studies with Glu298Asp and 4b/a polymorphisms were included. A allele of the 4b/a polymorphism increased the risk of end stage renal disease (ESRD) in ADPKD (odds ratio (OR) = 1.85, 95% confidence interval (CI) 1.17-2.94, P = 0.009). However, GG phenotype of Glu298Asp polymorphism neither decreased the ESRD risk (OR = 0.77, 95% CI 0.55-1.08, P = 0.13) nor affected the hypertension risk (OR = 1.04, 95% CI 0.66-1.66, P = 0.86). The GG phenotype carriers had later ESRD age compared with the T allele of Glu298Asp polymorphism (WMD = 2.39; 95% CI 1.32-3.46; P < 0.0001). Significant association was also found in Caucasians (WMD = 2.41; 95% CI 1.18-3.64; P = 0.0001). Subgroup analysis by gender indicated GG genotype carriers had older age of ESRD than T allele carriers in males (WMD = 4.51; 95% CI 3.95-5.08; P = 0.00001), but not in females. CONCLUSIONS: GG genotype of the Glu298Asp variant slowed the ESRD progression in ADPKD, while a allele carriers of the 4b/a variant increased the risk of ESRD. Variants of eNOS gene might play different roles in the ESRD progression in ADPKD.

Highly Selective Coextraction of H+/SO42- Using a Strapped Calix[4]pyrrole.

We report on the synthesis of a cage-type calix[4]pyrrole (1) bearing an additional basic pyridinebisthiazolamine group on the strap. The receptor in its protonated form shows strong affinity and selectivity for sulfate over a wide range of inorganic anions. With receptor 1 as a liquid-liquid extractant, H+/SO42- in the form of H2SO4 is almost quantitatively extracted from an aqueous solution containing HNO3 at a high concentration to CH2Cl2 in a recyclable manner.

Tauroursodeoxycholic acid inhibits TGF-ß1-induced renal fibrosis markers in cultured renal mesangial cells by regulating endoplasmic reticulum stress.

Chronic kidney disease (CKD) has a worldwide prevalence of higher than 10% with an increasing mortality rate. As it involves the deterioration of renal function, it represents a serious risk to human health and, if left untreated, significantly lowers the quality of the patient's life. CKD is characterized by renal fibrosis. Studies have shown that transforming growth factor ß1 (TGF-ß1), a key driving factor of renal fibrosis, is closely related to the activation of renal fibrosis pathways such as endoplasmic reticulum stress (ERS). Tauroursodeoxycholic acid (TUDCA), an endogenous bile acid derivative, can effectively inhibit endogenous ERS. Here, we explored the effects and actions of TUDCA on renal fibrosis by establishing a renal mesangial cell (RMC) model. The RMC was stimulated with TGF-ß1, and PCR and western blotting were used to detect the expression of ERS-related chaperone proteins and fibrotic indicators. The expression of glucose-regulated protein 78 (GRP78) was silenced in RMC cells to investigate the role of GRP78 in renal fibrosis. Finally, PCR and western blotting were used to detect the effects of TUDCA on the expression of GRP78, C/EBP homologous protein (CHOP), α-smooth muscle actin (α-SMA), and fibronectin (FN) in the TGF-ß1-stimulated RMCs. The results showed that TUDCA significantly downregulated TGF-ß1-induced levels of GRP78, CHOP, α-SMA and FN in RMCs. In addition, downregulation of GRP78 inhibited the expression of FN and α-SMA in the RMCs. In conclusion, downregulation of GRP78 and CHOP expression is one of the mechanisms by which TUDCA inhibits TGF-ß1-induced renal mesangial cell fibrosis.

Primary hepatic paraganglioma mimicking hepatocellular carcinoma: a case report.

Background: Primary hepatic paraganglioma (HPGL) originates from sympathetic nervous tissue in the liver. It is one of an exceedingly rare kind of sympathetic paragangliomas. The radiological features and clinical characters of HPGL can be easily confused with hepatocellular carcinoma (HCC). We present a case of HCC that was preoperatively diagnosed as hepatic paraganglioma, however, was pathologically verified as hepatic paraganglioma after surgery. Case Description: The present case reported a 47-year-old female with a very rare HPGL without any clinical symptoms, except for hyper menorrhagia and paroxysmal hypertension. The Spiegelman lobe of the liver underwent hepatic magnetic resonance imaging, which revealed a 3.2×3.8 cm mass, with uneven arterial phase wash-in and rapid portal and delayed phase wash-out. According to the imaging results, the patient was first diagnosed with hepatocellular carcinoma, and a radical hepatectomy was performed. However, the blood pressure of the patient displayed dramatic changes when the tumor was stimulated in operation. There were no substantial abnormalities found in the bilateral renal and adrenal glands. Therefore, we presumed that the tumor was related to functional pheochromocytoma. The tumor tissue was shown to be positive for chromogranin A, synaptophysin, CD56, and vimentin by immunohistochemical analysis. As a result, the patient was diagnosed with HPGL after this pathologic evaluation. Conclusions: There are several similarities between HPGL and HCC. For the treatment of hepatic paraganglioma, surgical excision is the recommended practice. Although the majority of paragangliomas are benign, long-term monitoring is required to differentiate benign from malignant paragangliomas.

Effect of location of the His-tag on the production of soluble and functional Buthus martensii Karsch insect toxin.

The low yield and poor folding efficiency in vivo of soluble and active recombinant cysteine-rich proteins expressed in Escherichia coli are a major challenge for large-scale protein production and purification. Expression vectors containing Buthus martensii Karsch insect toxin (BmK IT) fused to the C terminus of the intein Ssp DnaB were constructed in an attempt to overcome this problem. Following purification and intein self-cleavage, the fusion protein His(6)-intein-IT produced insoluble BmK IT, while intein-IT-His(6) generated soluble and properly folded BmK IT. This result indicated that the positioning of the His(6) tag has a key role in the production of soluble and functional BmK IT.

[Expression of PKD1 and PKD2 transcripts and proteins and its significance in different types of kidney tissues and kidney lines].

OBJECTIVE: To investigate the expression and function of PKD1 and PKD2 in different kidney tissues and cell lines. METHODS: Immunoprecipitation, Western blotting, In situ hybridization and immunohistochemical staining methods were used to observe the expression of PKD1 mRNA and PKD2 mRNA and their protein abundance in different kidney tissues and cell lines. RESULTS: Coordinate expressions of PKD1 and PKD2 were found in all kidney tissues and cell lines. Distribution of PKD1 mRNA and PKD2 mRNA and their protein polycystin-1 and polycystin-2 in normal human adult kidney tissue were mainly expressed in the medullary collecting ducts and distal tubules. Positive staining was also found in the majority of cyst-lining epithelial cells of PKD1 cystic kidney tissue, PKD1 cyst-lining epithelia cell line and LLC-PK1. The expression level of them in cystic epithelia of ADPKD kidney tissue was much higher than that in adult renal tubules (P < 0.01). CONCLUSIONS: Similar expression pattern of PKD1 and PKD2 and their different tissue distribution in different kidney tissues show that the molecular mutuality of PC-1 and PC-2 might be the base of their functional correlation. Polycystins might play an important role in the maintenance of tubular architecture.

A new method of preventing bile duct injury in laparoscopic cholecystectomy.

AIM: Of all the complications of laparoscopic cholectecystomy, bile duct injury (BDI) is the most serious complication. The prevention of injury to the common bile duct (CBD) remains a significant concern in laparoscopic cholecystectomy (LC). Different kinds of methods have been advanced to avoid this injury but no single method has gained wide acceptance. Because of various limitations of current methodologies we began a study using cold light illumination of the extrahepatic biliary system (light cholangiography LCP) to better visualize this area and thereby reduce the risk of bile duct injury. METHODS: Thirty-six patients with cholelithiasis were divided into two groups. Group I (16 cases) received LCP and group II (20 cases) received methelenum coeruleum cholangiography (MCCP). In group I cold light was used to illuminate the common bile duct by leading an optical fiber into the common duct with a duodenoscope at the time of LC. The light coming from the fiber in the CBD could clearly illuminate the location of CBD and hepatic duct establishing its location relative to the cystic duct. This method was compared with the dye injection technique using methelenum coeruleum. RESULTS: In group I thirteen cases were successfully illuminated and three failed. The cause of three failed cases was due to the difficulty in inserting the fiber into the ampulla of Vater. No complications occurred in the thirteen successful cases. In each of these successful cases the location of the common and hepatic ducts was clearly seen differentiating the ductal system from surrounding anatomy. In ten cases both the left and right hepatic ducts could be seen and in three only the right hepatic ducts were seen. In four of the thirteen cases, cystic ducts were also seen. In group II, eighteen of the twenty cases were successful. The location of extrahepatic ducts became blue differentiating the ductal system from surrounding anatomy. Two cases failed due to a stone obstructing the cystic duct, and extravisation of the dye turned the entire area blue. LCP showed the common and hepatic ducts more clearly than MCCP. CONCLUSION: LCP is the only technique that can clearly and directly show the location of the extrahepatic biliary system and may be useful in selecting cases of uncertain anatomy in the prevention of bile duct injury.

Detection of Haemophilus parasuis isolates from South China by loop-mediated isothermal amplification and isolate characterisation.

Haemophilus parasuis is the etiological agent of Glässer's disease, which is characterised by fibrinous polyserositis, meningitis and polyarthritis, causing severe economic losses to the swine industry. In this study, a loop-mediated isothermal amplification (LAMP) test was developed to improve the specificity, facility and speed of diagnosis of H. parasuis isolates. The LAMP assay rapidly amplified the target gene within 50 min incubation at 63 °C in a laboratory water bath. The LAMP amplicon could be visualised directly in the reaction tubes following the addition of SYBR Green I dye. The detection limit of this LAMP method was 10 CFU/mL, which was 10 times more sensitive than the earlier 16S rRNA polymerase chain reaction (PCR) test conducted by Oliveira, Galina and Pijoan (2001), and no cross-reactivity was observed from other non-H. parasuis strains. This LAMP test was evaluated further on 187 clinical specimens from pigs suspected of being infected with H. parasuis. Forty-three were found positive by bacterial isolation of H. parasuis, as well as by the 16S rRNA PCR and LAMP tests. The 43 H. parasuis isolates were classified into 9 serovars and had 37 genetic patterns when analysed by pulsed-field gel electrophoresis (PFGE). This displayed that various H. parasuis serovars and genotypes were widely distributed in South China. Therefore, the speed, specificity and sensitivity of the LAMP test, the lack of a need for expensive equipment, and the visual readout showed great potential for a correct clinical diagnosis of H. parasuis in favour of controlling Glässer's disease.

Scientific publications in nephrology and urology journals from Chinese authors in East Asia: a 10-year survey of the literature.

BACKGROUND: Diseases of the kidneys and genitourinary tract are common health problems that affect people of all ages and demographic backgrounds. In this study, we compared the quantity and quality of nephrological and urological articles published in international journals from the three major regions of China: the mainland (ML), Hong Kong (HK), and Taiwan (TW). METHODS: Nephrological and urological articles originating from ML, TW, and HK that were published in 61 journals from 1999-2008 were retrieved from the PubMed database. We recorded the numbers of total articles, clinical trials, randomized controlled trials, case reports, impact factors (IF), citations, and articles published in the leading general-medicine journals. We used these data to compare the quantity and quality of publication output from the three regions. RESULTS: The total number of articles increased significantly from 1999 to 2008 in the three regions. The number of articles from ML has exceeded that from HK since 2004, and surpassed that from TW in 2008. Publications from TW had the highest accumulated IF, total citations of articles, and the most articles published in leading general-medicine journals. However, HK publications had the highest average IF. Although ML produced the largest quantity of articles, it exhibited the lowest quality among the three regions. CONCLUSION: The number of nephrological and urological publications originating from the three major regions of China increased significantly from 1999 to 2008. The annual number of publications by ML researchers exceeded those from TW and HK. However, the quality of articles from TW and HK was higher than that from ML.

Prevention of Biliary Duct Injury in Laparoscopic Cholecystectomy Using Optical Fiber Illumination in Common Bile Duct.

BACKGROUND: Biliary duct injury (BDI) is one of the most common complications in laparoscopic cholectecystomy (LC), in this study, we have tried to place an illuminating optical fiber via endoscopy in the CBD during LC, the biliary duct anatomy can be clearly delineated, thus CBD injury is avoided. METHODS: Sixteen patients with chronic cholecystitis or/and cholelithiasis from February 2007 to June 2008 were performed LC with placement of optical fiber in CBD, the fiber with cold light illuminates the whole extrahepatic biliary system. Three 6-mm titanium clips were applied to the soft tissue surrounding the hepatic duct, CBD and the cystic duct confluence with CBD, respectively; one titanium clip was applied to the surface of cystic duct near the infundibulum of gallbladder. The cytic duct, CBD and common hepatic duct were clearly identified and delineated in the operating field and LC was performed. RESULTS: All the 16 patients were performed LC using this procedure successfully, there were no LC-related complications, nor complications related to endoscopic retrograde cholangiopanceatography (ERCP). CONCLUSIONS: The endoscopically placed optical fiber in the CBD can clearly identify the CBD, Calot's triangle and the common hepatic duct, this can reduce the bile duct injury in LC and imporve the safety of LC.

Meta-analysis of Tripterygium wilfordii Hook F in the immunosuppressive treatment of IgA nephropathy.

OBJECTIVE: Numerous Chinese patients with IgA nephropathy (IgAN) have benefited from Tripterygium wilfordii Hook F (TwHF) from two decades ago. However, to date there is no systematic evaluation of this remedy for IgAN. METHODS: We conducted a meta-analysis of all eligible randomized clinical trials (RCTs) to assess the effect of TwHF on IgAN for the first time. In August 2009 a systematic search was performed among eight electronic databases. Review Manager (RevMan) version 5.0 was used. RESULTS: (i) Four eligible RCTs with 188 participants were included; (ii) The validities of included RCTs were generally acceptable; (iii) TwHF brought about a favorable increase in complete remission (CR) (RR 1.53, 95%CI 1.09 to 2.16, I(2)=12%) and total remission (TR) (RR 1.27, 95%CI 1.08 to 1.48, I(2)=0%) compared with non-TwHF treatment; and this result was further confirmed by intention-to-treat analysis; (iv) Exploiting subgroup meta-analysis, TwHF led to significantly greater improvements of IgAN with non-nephrotic proteinuria with regard to the increase of CR (RR 1.80, 95%CI 1.21 to 2.68, I(2)=0%) and TR (RR 1.32, 95%CI 1.11 to 1.57, I(2)=0%), and decrease of urinary proteinuria excretion (UPE) (MD -467.41 mg/24h, 95%CI -633.99 to -300.82, I(2)=0%). Meanwhile, the renal function was well preserved (MD -2.66 µmol/L, 95%CI -9.26 to 3.94, I(2)=0%). Conclusion Although the results of this meta-analysis should be interpreted with caution and warrant further investigation, TwHF was certainly a valuable and promising immunosuppressive remedy for IgAN, which was in accordance with the accruing evidence from numerous large clinical and experimental studies.

[Molecular analysis of an avian influenza virus isolate of H5N2 subtype from parrot].

In 2005, an avian influenza virus stain was isolated from Parrot in Guangdong, which was then genotyped as H5N2 subtype and designated as A/Parrot/Guangdong/268/2005. According to the current OIE definition on the low-pathogenicity of avian influenza virus, the strain was recognized as a low pathogenic avian influenza virus due to the presence of one basic amino acid residue at the HA cleavage site. Some molecular characteristics of the virus, such as potential glycosylation sites in HA and NA, receptor binding sites of HA, and drug resistance site of NA, showed no variations. To analyze molecular evolution of this strain, we selected the sequences of H5N2 subtype AIVs from GenBank and established the phylogenetic trees. Our results indicated that this strain shared the highest homologies with the H5N2 LPAI isolate A/Pheasant/NJ/1355/1998-like. Phylogenic analysis revealed the isolate, together with A/Chicken/Pennsylvania/1/1983 (H5N2), belonged to America lineages and clustered with A/Pheasant/NJ/1355/1998-like.

[Soluble expression, purification and characterization of Bm K IT in Escherichia coli by intein-mediated system].

To produce recombinant Buthus martensii Karsch insect toxin (BmK IT), BmK IT cDNA which fused a hexahistidine sequence at the C-terminus by PCR was inserted into pTWIN1 expression vector fused in frame with an upstream Ssp DnaB intein gene. The expression plasmid was transformed into E. coli BL21 (DE3) strain and protein expression was induced by IPTG. The CBD-Intein-BmK IT(his6) fusion protein was purified from cell lysates using Ni-NTA resin affinity chromatography. The intein was removed from fusion protein by on-column intein-mediated cleavage. BmK IT(his6) was purified through Superdex 75 gel chromatography to more than 95% homogeneity. The purified protein has both correct secondary structure and insecticidal activity.

Potential biochemical therapy of glioma cancer.

Glioma is a highly invasive, rapidly spreading form of brain cancer that is resistant to surgical and medical treatment. The recent progresses made in intracellular and ion channels of glioma cells provide a potential new approach for biochemical therapy of brain tumor. In this paper, we reviewed clinical data on chemotherapy by temozolomide and results from new studies on voltage-gated potassium channels, large-conductance Ca(2+)-activated K(+) channels, volume-activated chloride channels, glioma-specific chloride channel and their modulators. These new findings may represent future directions for brain tumor studies and treatment.

Expression of an insect excitatory toxin, BmK IT, from the scorpion, Buthus martensii Karsch, and its biological activity.

An insect excitatory toxin from Buthus martensii Karsch (BmK IT) was cloned into the expression vector, pTWIN1, and expressed into Escherichia coli BL21 (DE3) host cells. The soluble fusion expression of CBD-intein-BmK IT was obtained. The recombinant BmK IT was purified by two anion-exchange chromatography columns and one gel chromatography column. Bioassays were carried out to verify the toxicity of this recombinant toxin. At the end of a 96 h experimental period, 83% of cotton bollworm larvae were killed with an LT(50) value of 58-62 h. Furthermore, the average weight of larvae fed on BmK IT-containing media was approx 4% of that of the control groups. The results indicate that the expressed and purified recombinant BmK IT has biological activity.