Cutaneous Fistula ; Digestive System Abnormalities ; therapy ; Endoscopy, Gastrointestinal ; methods ; Female ; Humans ; Infant ; Treatment Outcome

To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.
Animals ; Biological Transport ; Cimetidine ; pharmacology ; DNA, Complementary ; Dogs ; Drug Interactions ; Humans ; Madin Darby Canine Kidney Cells ; Metformin ; pharmacology ; Organic Cation Transport Proteins ; genetics ; metabolism ; Transfection

Objective:To examine the expression ofphospholipase A2 receptor (PLA2R) in renal tissues and the level of anti-PLA2R antibody in serum in patients with idiopathic membranous nephropathy (IMN) and secondary membranous nephropathy (SMN),and to evaluate their diagnostic value in IMN.Methods:A total of 73 patients,who were diagnosed between May,2014 and February,2015 in the Department of Nephrology of the Second Xiangya Hospital,Central South University,were divided into three groups:an IMN group (n=48),an SMN group (n=17) and a minimal change disease group (n=8) according to the renal biopsy.PLA2R expression in renal tissues and the level of antiPLA2R antibody in serum were detected by indirect immunofluorescence technique.Results:The positive rate and fluorescence intensity for PLA2R in the renal tissues in the IMN group were higher than those in the SMN group (91.7％ in the IMN group vs 29.4％ in the SMN group,P＜0.05),while the positive rate and serum level for anti-PLA2R antibody in the IMN group were higher than those in the SMN group (85.4％ in the IMN group vs 29.4％ in the SMN group,P＜0.05);the expression of PLA2R in renal tissues and the serum level for anti-PLA2R antibody were not detected in the minimal change disease group,The serum level of anti-PLA2R antibody was positively correlated with 24 h urine protein (r=0.432,P＜0.01) and negatively correlated with serum albumin (r=-0.307,P＜0.05).Conclusion:The expression of PLA2R in renal tissues and the serum level of anti-PLA2R antibody might be potential markers for diagnosis oflMN.

Objective To observe the protein and mRNA expression of thyroid-stimulating hormone receptor (TSHR) in mammary gland tissue of lactating rats,and to explore iodine uptake mechanism.Methods Eighty adult Wistar rats (60 female and 20 male),weighting 210-250 g were selected.All female Wistar rats were randomly divided into 6 groups according to their body mass:normal non-pregnant group,lactating for 5-,10-,15-and 20-day groups and weaning for 5 days group,10 rats in each group.All rats were fed with conventional fodder and tap water freely.In addition to the normal non-pregnant group,other five groups of female and male rats were mated at 3 ∶ 1,respectively.Then the rats in all groups were killed on the 5th,10th,15th and 20th day after lactation and on the 5th day after weaning to get the mammary gland tissue.The protein and mRNA expression of TSHR were determined by immunohistochemical staining and real-time quantitative PCR.Results TSHR protein was expressed in mammary acinar and ductal epithelial cytoplasm.The expression of TSHR in mammary gland showed significant differences between groups (x2 =14.612,P ＜ 0.05),the staining intensity of mammary gland tissue in normal non-pregnant rats(weak,n =4; moderate,n =6) was weaker than that of lactating for 5 days(weak,n =2; moderate,n =3; strong,n =5) and 10 days groups(barely detectable,n =1;moderate,n =4; strong,n =5; x2 =4.113,5.250,all P＜ 0.05).The expression of TSHR mRNA in mammary gland showed significant differences between groups(F=20.488,P ＜ 0.05); the expression of TSHR mRNA in lactating for 10 days group(0.31 ± 0.06) was higher than that of lactating for 5 days group(0.22 ± 0.04,P ＜ 0.01),and the expression of lactating for 15 days group (0.16 ± 0.08) was significantly lower than that of lactating for 5 days group (P ＜ 0.05).Conclusions TSHR is widely expressed in mammary gland of lactating rats.The iodine uptake of mammary gland is enhanced in early lactation period when the body may be more susceptible to iodine deficiency,therefore iodine should be supplemented reasonably.

BackgroundThe influence of glaucoma on the quality of life in patients is of increasing concern for ophthalmologists in recent years. However,some studies demonstrated that different types of questionnaires about quality of life have various disadvantages. Therefore, to accurately and fully assess the influencing factors of quality of life in glaucoma patients is very important. ObjectiveThe present study was to survey the self-reported visionrelated quality of life(QOL) in glaucoma patients by means of utility analysis and to tentatively analyze its influencing factors. Methods Patients with glaucoma were recruited from a single tertiary ophthalmic department. Standard face-to-face interviews were conducted. Utility values of linear rating scale ( RS ) and time trade-off ( TTO ) were calculated to evaluate the self-reported vision-related QOL of the patients. The correlations of the utility values with the patients' general and ophthalmologic characteristics were also analyzed. This survey was approved by the Ethic Committee of Beijing Tongren Hospital. Oral informed consent was obtained from the subjects before the study.ResultsA cross-sectional study was designed. A total of 86 glaucoma patients were enrolled in this study with 62 male and 24 female, with a mean age of 44. 67 years old. The mean utility values measured by RS and TTO were 0. 62± 0. 19 and 0. 77 ± 0. 12, respectively, and no evidential correlation was found between these two values ( r =0. 074, P=0. 499 ). The RS value was associated with daily visual acuity,mean deviation(MD) of visual field and the history of trabeculectomy. Neither daily visual acuity nor MD showed a significant correlation with the TTO value. Age, work status and educational background contributed to higher utility value for the TTO method. After adjusting for age, work status and educational level,patients with visual acuity in the worse-seeing eye better than 0. 3 showed a higher TTO value than those with less than 0. 3. Conclusions Utility analysis possesses the advantages of convenience and sensitivity. RS utility value is easily affected by the Objective visual status and surgery history in glaucomatous patients,which reflects the subjective assessment of patients'visual quality. However, TTO value is primarily associated with age,work status and education level rather than visual function in glaucoma patients,which is therefore subjective assessment of the disease-related quality of life. These Results indicate that visual function impairment is not a determining factor for the QOL of glaucoma patients.

Two hundred thirty specimens of wild birds were collected from some areas in Heilongjiang Province during the period of 2003~2004, including two batches of specimens collected randomly from a same flock of mallards in Zhalong Natural Reserve in August and December, 2004, respectively. Primary virus isolation and identification for avian influenza virus (AIV) and Newcastle disease virus (NDV) were performed. The results showed that only two specimens of young mallards collected from Zhalong Natural Reserve in August, 2004 were positive to AIV (isolation rate 0.9%), and one strain (D57) of these two virus isolates was identified to be H9 subtype by hemagglutination inhibition test. Meanwhile, the two batches of blood serum samples of mallards from Zhalong were also examined for antibodies against AIV and NDV. Among 38 blood serum samples collected in August, antibodies against the hemagglutinin of H1, H3, H5, H6 and H9 subtypes of AIV were found in 1, 0, 2, 0 and 8 samples, respectively; and 11 samples were found with antibody against NDV. Whereas the NDV isolation in both two batches of specimens of mallard was negative, all of the 32 blood serum samples collected in December were negative for antibodies against AIV and NDV.
Animals ; Animals, Wild/*virology ; Antibodies, Viral/isolation&purification ; Birds/virology ; China/epidemiology ; Hemagglutination Tests ; Influenza A virus/*isolation&purification ; Influenza in Birds/epidemiology/immunology/*virology ; Newcastle Disease/epidemiology/immunology/*virology ; Newcastle disease virus/*isolation&purification ; Reverse Transcriptase Polymerase Chain Reaction

To investigate the difference in absorptive of tetrahydropalmatine (THP) and l-tetrahydropalmatine (l-THP) in rat intestine as well as the mechanism of the absorption of THP, in situ single pass perfusion model was used and the concentration of THP in perfusate was determined by HPLC. The absorption rate constant (k(a)) and effective permeability values (P(eff)) of THP had no significant difference (P > 0.05) at concentration of 8, 16 and 32 microg x mL(-1) in perfusion or in four different regions of intestine of rat (duodenum, jejunum, ileum, colon). The absorption of l-THP and THP in jejunum had significant difference (P < 0.05). The k(a) and P(eff) of THP increased obviously when verapamil was co-perfused with THP, while those of l-THP were not influenced by verapamil. The absorption of THP in intestine showed the passive diffusion process, and without a special absorption region. The stereoselective absorption difference may result from stereoselective combination of P-glycoprotein with d-THP.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Animals ; Berberine Alkaloids ; chemistry ; pharmacokinetics ; Colon ; metabolism ; Duodenum ; metabolism ; Female ; Ileum ; metabolism ; Intestinal Absorption ; drug effects ; Jejunum ; metabolism ; Male ; Perfusion ; Permeability ; Rats ; Rats, Sprague-Dawley ; Sex Factors ; Stereoisomerism ; Verapamil ; pharmacology

OBJECTIVETo evaluate the long-term effects of delayed hyperbaric oxygen (HBO) therapy on neonatal rats with hypoxic-ischemic brain injury (HIBD).

METHODPostnatal 7 days newborn rats (n = 52) were randomly set to three groups: control (n = 18, sham operation), HIBD (n = 17), or HBO (n = 17). Pups in the HBO group were subjected to hyperbaric oxygen treatment with 2 atmosphaera absolutus, 5 x 30 min at a 24 h intervals since 48-72 h after the HIBD model. All the animals were tested for the spatial learning and memory ability in the Morris water maze from postnatal days 37 to 41. At day-42, rats were decapitated and the brains were analyzed for morphological and histological changes, including brain shapes and weights, survival neurons, percentage of AchE positive area and NOS positive neurons in hippocampal CA1 region.

RESULTSRats in HBO and HIBD groups displayed significant morphological and histological damages, as well as severe spatial learning and memory disability. The average escape latency of Morris water maze in HBO group [(56 +/- 23) s] and HIBD group [(56 +/- 22) s] were longer than the control [(23 +/- 16) s] (P < 0.05). The swimming time in HBO group [(30 +/- 5) s] and HIBD group [(29 +/- 6) s] were shorter than the control [(51 +/- 5) s] (P < 0.05). The swimming length in HBO group [(572 +/- 92) cm] and HIBD group [(548 +/- 92) cm] were shorter than the control [(989 +/- 101) cm] (P < 0.05). The weight of left brains in HBO group [(598 +/- 46) mg] and HIBD group [(601 +/- 59) mg] were lighter than the control [(984 +/- 18) mg] (P < 0.05). The survival neurons of hippocamal CA1 region in HBO group [(97 +/- 27)/mm] and HIBD group [(100 +/- 27)/mm] were less than the control [(183 +/- 8)/mm] (P < 0.05). The percentage of AchE-positive fibers in HBO group [(18.4 +/- 2.2)%] and HIBD group [(18.5 +/- 2.2)%] were less than the control [(27.5 +/- 2.2)%,] (P < 0.05). NOS-positive neurons in HBO group [(21 +/- 5)/mm(2)] and HIBD group [(19 +/- 4)/mm(2)] were also less than the control [(34 +/- 6)/mm(2)] (P < 0.05).

CONCLUSIONDelayed HBO therapy resulted in no protection against either HIBD-induced brain morphological and histological deficits or spatial learning and memory disability.

Acetylcholinesterase ; analysis ; Animals ; Animals, Newborn ; Brain ; pathology ; Female ; Hippocampus ; pathology ; Hyperbaric Oxygenation ; Hypoxia-Ischemia, Brain ; drug therapy ; pathology ; Male ; Maze Learning ; Nitric Oxide Synthase ; analysis ; Rats ; Time

OBJECTIVETo study the validity of criteria currently used in China for the classification of symmetric small for gestational age infants (SGA) as compared with its definition.

METHODSThis study included 417 inpatients diagnosed as SGA in authors' hospital from January 1998 to June 2002. Symmetric SGA was diagnosed by the following three criteria: (1) the Ponderal Index (PI), (2) the crown-heel length-to-head circumference ratio (BL/HC) issued in Chin J Pediatr (1988;26:164 - 165), as well as (3) the SGA definition. The definition criterion was considered as the "gold standard". The sensitivity, specificity, false positive and negative values, positive and negative predictive values, exact agreement ratio, diagnosis index, and Cohen's Kappa value were used to evaluate the validity and agreement of the methods of PI and BL/HC. Receiver Operating Characteristic (ROC) analysis was used to evaluate the validity of the diagnosis.

RESULTSOf 417 SGA infants, 376 (90.17%), 376 (90.17%) and 187 (44.84%) subjects were diagnosed as symmetric type with PI, BL/HC and the definition criteria, respectively. (2) The agreement rate and Kappa value between PI and BL/HC was 80.82% and -0.093 (SEM 0.026), respectively. And the agreement rates between PI or BL/HC and the definition criterion were 49.88% and 50.84%, respectively. As compared with the definition criterion, the PI and BL/HC methods had sensitivities of 91.8% - 96.4%, specificities of 9.3% - 25.9%, positive predictive values of 45.8% - 51.1%, negative predictive values of 72.7% - 82.8%, diagnosis indices of 4.9% - 17.7% and Kappa values of 0.070 - 0.167. (3) The areas under the ROC curves in full-term and preterm infants by PI method were 0.635 (95% CI, 0.573 - 0.697) and 0.698 (95% CI, 0.622 - 0.725), respectively. PI cutoffs at 2.47 in full-term SGA, at 2.43 in preterm SGA, and BL/HC cutoff at 1.43 produced the maximum diagnosis indices that were 24.7%, 39.6% and 33.7%, respectively. When the PI at 2.50 (full-term), PI at 2.31 (preterm) and BL/HC values at 1.46, the sensitivity closed mostly to the specificity. The sensitivities and specificities in full-term and preterm infants were 59.4% and 59.3%, 65.3% and 65.5%, and 66.3% and 65.5%, respectively.

CONCLUSIONIn the classification of SGA, the results showed a poor agreement between PI or BL/HC and the definition criterion. The results suggested that the current cutoffs of PI and BL/HC might not be appropriate for the diagnosis of symmetric SGA. Low AUC suggested that PI and BL/HC could not give a valid diagnosis at any cutoffs.

Anthropometry ; methods ; Birth Weight ; Body Height ; China ; Female ; Gestational Age ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Reproducibility of Results ; Sex Factors

OBJECTIVETo study the genetic heterogeneity of autosomal dominant polycystic kidney disease (ADPKD) in Chinese.

METHODSUsing polymerase chain reaction (PCR) and non-denatured polyacrylamide gel electrophoresis, the authors analyzed eight microsatellite markers closely linked to PKD1 or PKD2 genes respectively in a Chinese ADPKD family.

RESULTSSeven informative markers were found in this family, including KG8, SM6, CW4 and CW2 which are tightly linked to PKD1, and D4S1563, D4S414 and D4S423 which are linked to PKD2. After the process of genotyping, the haplotypes were estimated with Cyrillic 2.0, and the linkage-based analysis suggested that the disease is not linked to PKD1 other than PKD2.

CONCLUSIONIn China this non-PKD1 family is the second one, but it is the first reported PKD2 family showing the genetic heterogeneity of ADPKD in Chinese. In the family the affected mother transmits the disease and the affected members' phenotypes are eterogeneous. In addition, the existing "anticipation" and the presence of the disease in a child of this family suggest that non-PKD1 linked families may have early-onset of the disease in child.

Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; China ; Family Health ; Female ; Haplotypes ; genetics ; Humans ; Microsatellite Repeats ; genetics ; Middle Aged ; Pedigree ; Polycystic Kidney, Autosomal Dominant ; ethnology ; genetics ; Polymerase Chain Reaction ; TRPP Cation Channels ; genetics