Correlations Between Single Nucleotide Polymorphisms, Cognitive Dysfunction, and Postmortem Brain Pathology in Alzheimer's Disease Among Han Chinese / 神经科学通报·英文版

In this study, the distribution of five Alzheimer's disease (AD)-related single nucleotide polymorphisms (SNPs) in the Han population was examined in combination with the evaluation of clinical cognition and brain pathological analysis. The associations among SNPs, clinical daily cognitive states, and postmortem neuropathological changes were analyzed in 110 human brains from the Chinese Academy of Medical Sciences/Peking Union Medical College (CAMS/PUMC) Human Brain Bank. APOE ε4 (OR = 4.482, P = 0.004), the RS2305421 GG genotype (adjusted OR = 4.397, P = 0.015), and the RS10498633 GT genotype (adjusted OR = 2.375, P = 0.028) were associated with a higher score on the ABC (Aβ plaque score, Braak NFT stage, and CERAD neuritic plaque score) dementia scale. These results advance our understanding of the pathogenesis of AD, the relationship between pathological diagnosis and clinical diagnosis, and the SNPs in the Han population for future research.

Expression pattern of congenital chloride diarrhea pathogenic gene Slc26a3 in the reproductive tract of male rodents / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To determine the expression pattern of Slc26a3 gene in reproductive tract of male rodents to clarify whether the expression pattern is related to the subfertility observed in congenital chloride diarrhea (CLD) disease.</p><p><b>METHODS</b>The expression of Slc26a3 in mouse and rat epididymis has been studied with immunohistochemistry and Western blotting. Its developmental expression pattern in rat testis was detected by Western blotting, while both of immunofluorescence and Western blotting were used to localize the expression of Slc26a3 in mouse sperms. The potential change of Slc26a3 expression in CFTR (cystic fibrosis transmembrane conductance regulator) knockout mice and CFTR mutant mice was also detected with Western blotting.</p><p><b>RESULTS</b>The expression level of Slc26a3 gradually decreased along epididymis from its caput to corpus, then to its cauda part. This gradually decreasing expression pattern was also found in rat testis during development. Slc26a3 was localized mainly on the trunk of mouse sperm tail. In the testis and epididymis of CFTR knockout mice and CFTR mutant mice, no significant change of Slc26a3 expression was found.</p><p><b>CONCLUSION</b>Slc26a3 is expressed in male reproductive tract, and its expression pattern is related to the function. Thus, the subfertility observed in CLD disease may be related to the important role of SLC26A3 in acid-base regulation of epididymis.</p>

Lysophosphatidic acid increases SLC26A3 expression in inflamed intestine and reduces diarrheal severity in C57BL/6 mice with dextran-sodium-sulfate-induced colitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Diarrhea is a common clinical feature of ulcerative colitis resulting from unbalanced intestinal fluid and salt absorption and secretion. The Cl(-)/HCO3(-) exchanger SLC26A3 is strongly expressed in the mid-distal colon and plays an essential role in colonic Cl(-) absorption and HCO3(-) secretion. Slc26a3 expression is up-regulated by lysophosphatidic acid (LPA) in vitro. Our study was designed to investigate the effects of LPA on SLC26A3 expression and the diarrheal phenotype in a mouse colitis model.</p><p><b>METHODS</b>Colitis was induced in C57BL/6 mice by adding 4% of dextran sodium sulfate (DSS) to the drinking water. The mice were assigned to LPA treatment DSS group, phosphate-buffered saline (PBS) treatment DSS group, DSS only group and untreated mice with a completely randomized design. Diarrhea severity was evaluated by measuring mice weight, disease activity index (DAI), stool water content and macroscopic evaluation of colonic damage. The effect of LPA treatment on Slc26a3 mRNA level and protein expression in the different groups of mice was investigated by quantitative PCR and Western blotting.</p><p><b>RESULTS</b>All mice treated with DSS lost weight, but the onset and severity of weight loss was attenuated in the LPA treatment DSS group. The increases in stool water content and the macroscopic inflammation score in LPA treatment DSS group were significantly lower compared to DSS control group or PBS treatment DSS group ((18.89±8.67)% vs. (28.97±6.95)% or (29.48±6.71)%, P = 0.049, P = 0.041, respectively and 2.67±0.81 vs. 4.5±0.83 or 4.5±0.54, P = 0.020, P = 0.006, respectively), as well as the increase in DAI (P = 0.004, P = 0.008, respectively). LPA enema resulted in higher Slc26a3 mRNA and protein expression levels compared to PBS-treated and untreated DSS colitis mice.</p><p><b>CONCLUSION</b>LPA increases Slc26a3 expression in the inflamed intestine and reduces diarrhea severity in DSS-induced colitis, suggesting LPA might be a therapeutic strategy in the treatment of colitis associated diarrhea.</p>

Effects of weile powder on bicarbonate transporters CFTR SLC26A3 and SLC26A6 in gastric ulcers of rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effects of Weile Powder (WLP) on bicarbonate transporters in rats with gastric ulcers, and to probe its functional mechanisms.</p><p><b>METHODS</b>The 48 SD rats were randomly divided into the normal control group, the model group, the low dose WLP group (at the daily dose of 0.075 g/mL), the middle dose WLP group (at the daily dose of 0.150 g/mL), the high dose WLP group (at the daily dose of 0.030 g/mL), and the ranitidine group (at the daily dose of 0.030 g/mL), 8 in each group. The gastric ulcer rat model was prepared by the glacial acetic acid cauterization method. Rats in each medication group were administered from the 2nd day of modeling. Rats were sacrificed after 14-day successive medication. The protein was extracted from the ulcer tissue. The protein expressions of solute carrier26A3 (SLC26A3)and solute carrier26A6 (SLC26A6) were detected using Western blot. The gastric ulcer and its peripheral tissue were sectioned. The changes of cystic fibrosis transmembrane conductance regulator (CFTR) were measured by immunofluorescence.</p><p><b>RESULTS</b>Compared with the model control group, the expression levels of SLC26A3 increased in the high dose WLP group and the ranitidine group with statistical difference (P < 0.05). The expression levels of SLC26A6 increased in the high and middle dose WLP groups and the ranitidine group with statistical difference (P < 0.05). The expression level of CFTR also obviously increased in the high and middle dose WLP groups (P < 0.01).</p><p><b>CONCLUSION</b>WLP could elevate the expression levels of SLC26A6, SLC26A3, and CFTR, increase the secretion of bicarbonate, thus protecting the gastric mucosa.</p>

Gene mutations and clinical manifestations in children with glycogen storage disease type Ib / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Glycogen storage disease type Ib (GSDIb) is caused by a deficiency of glucose-6-phosphate translocase (G6PT) activity due to SLC37A4 gene mutations. Most GSDIb patients have recurrent infections and inflammatory bowel disease, with poor prognosis. Detection of SLC37A4 gene mutations is of great significance for the diagnosis, subtyping and outcome prediction of GSD patients. This study aims to analyze SLC37A4 gene mutations in Chinese GSDIb patients and to investigate the relationship between its genotypes and clinical manifestations.</p><p><b>METHODS</b>All exons and their flanking introns of SLC37A4 gene in 28 Chinese children with a primary diagnosis of GSDIb were screened by PCR combined with direct DNA sequencing to detect SLC37A4 gene mutations.</p><p><b>RESULTS</b>Five SLC37A4 gene mutations were detected in 7 (25%) of the 28 children, i.e., p.Gly149Glu (9/13, 69%), p.Gly115Arg (1/13, 8%), p.Pro191Leu (1/13, 8%), c.959-960 insT (1/13, 8%) and c.870+5G>A (1/13, 8%).</p><p><b>CONCLUSIONS</b>In this study, c.959-960 insT is a novel mutation and p.Gly149Glu is the most common mutation. p.Gly149Glu may be associated with severe infections in children with GSDIb.</p>

Organic anion transporting polypeptide (oatp4a1) mRNA and protein expressions in high fat and over-fatigue impairing Pi rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study organic anion transporting polypeptide (OATP) superfamily member 4a1 (oatp4a1) mRNA expression in the Pi deficiency model rats, thus exploring its mechanism for transporting and transforming the dampness.</p><p><b>METHODS</b>Six SD rats of SPF grade were used to prepare over-fatigue impairing Pi model. Another 12 SD rats were randomly divided into the blank control group and the high fat diets group, 6 in each. The special binding tube was used for the over-fatigue impairing Pi model group on the odd day, 3 h each time. Then the rats were forced to swim in the cold water (10 degrees C +/- 1 degrees C) for 7 min on the even day, for 2 successive weeks. Rats in the model group and the blank control group were granulated feed for 12 weeks, while rats in the high fat group were fed with high fat diet for 12 weeks. All rats were free to take food and drink water. The mRNA and protein expressions of oatp4al were detected in the Fei, Pi, Gan, Shen, Wei, Xiaochang, and Dachang using Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blot.</p><p><b>RESULTS</b>Rats in the model group were idled together with lousy defecate and coarse skin. They ate and drank less, and lost body weight (P<0.05). They were consistent with clinical manifestations of Pi deficiency syndrome, indicating that the over-fatigue impairing Pi animal model was successfully established. Rats in the high fat group started to have poor appetite and languish spirit, move lazily and addict to sleep, have coarse, dark, and colorless hair 9 weeks later, indicating phlegm dampness syndrome. Compared with the blank control group, the average body weight increased in the high fat group at the 9th week (P<0.05). The oatp4a1 mRNA expressed in the Fei, Pi, Gan, Shen, Wei, Xiaochang, and Dachang. There was no statistical difference in the oatp4al mRNA expression among all tissues (P>0.05). The oatp4al mRNA expressions were higher in the Fei and Shen of the high fat group than in the Gan (P<0.05).</p><p><b>CONCLUSIONS</b>oatp4al might be one of the basic substances in the transportation and transformation of phlegm dampness. Of them, Fei, Shen, and Dachang might play important roles in the transportation and transformation of phlegm dampness.</p>

Detection and sequencing of plasmid encoded tetracycline resistance determinants (tetA and tetB) from food-borne Bacillus cereus isolates

OBJECTIVE@#To investigate the detection and sequencing of plasmid encoded tetracycline resistance genes (tetA and tetB) from food-borne and standard strains of Bacillus cereus (B. cereus).@*METHODS@#A PCR was carried out to detect the tetracycline resistance genes (tetA and tetB) in food-borne B. cereus strains and the amplified products were sequenced.@*RESULTS@#The phenotypic resistance against tetracycline was observed in 39 of the 118 food-borne isolates and two reference strains (MTCC 430 and MTCC 1307) of B. cereus. Among the phenotypically resistant isolates, tetA was detected in 36 food-borne isolates and two reference strains (MTCC 430 and MTCC 1307), whereas, tetB was detected in 12 food-borne isolates and MTCC 1307 strain.@*CONCLUSIONS@#A close association was therefore found between phenotypic resistance against tetracycline and presence of tetracycline resistance genes. The tetA and tetB gene fragments were amplified, purified and sequenced. The gene sequences of the isolates studied herein were found similar to tetA and tetB gene sequences of other bacteria available in NCBI. The occurrence of tetA and tetB genes in B. cereus indicate the horizontal transfer of antibiotic resistance determinants from other bacteria into B. cereus. The transfer of these resistant determinants to other potentially pathogenic bacteria may be a matter of great concern.

Primary superficial small cell neuroendocrine carcinoma of esophagus: clinicopathologic and immunohistochemical analysis of 15 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic and immunohistochemical features of primary superficial esophageal small cell neuroendocrine carcinoma (ESCNC).</p><p><b>METHODS</b>The clinical, pathologic and immunohistochemical features were retrospectively analyzed in 15 cases of superficial ESCNC. An immunohistochemical study for chromogranin A, neuron-specific enolase, synaptophysin, CD56, TTF-1, 34βE12, AE1/AE3, and CK10/13 was performed.</p><p><b>RESULTS</b>Superficial ESCNC accounted for 4.8%(15/312) of all cases of superficial carcinoma of the esophagus encountered during the same period. The median survival time was 19 months and the mean survival time was 23.7 months after diagnosis. The one, two and five-year survival rates were 10/15, 5/15 and 1/15, respectively. The immunophenotypic profile was as follows: neuron-specific enolase (15/15), synaptophysin (15/15), AE1/AE3 (15/15), CD56 (14/15), TTF-1 (9/15), chromogranin A (8/15), 34βE12 (1/15) and CK10/13 (0/15).</p><p><b>CONCLUSIONS</b>Superficial ESCNC is a rare and aggressive malignant tumor with poor prognosis. Surgical resection coupled with post-operative chemoradiotherapy is the mainstay of treatment. The immunohistochemical study is valuable in pathologic diagnosis and differential diagnosis.</p>

Retroperitoneal laparoscopic radical nephrectomy in the treatment of renal cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the safety and efficacy of retroperitoneal laparoscopic radical nephrectomy in the treatment of renal cancer.</p><p><b>METHODS</b>The clinical data of 53 cases who underwent retroperitoneal laparoscopic radical nephrectomy were analyzed retrospectively.</p><p><b>RESULTS</b>Fifty-two cases achieved successful retroperitoneal laparoscopic radical nephrectomy, a conversion to open surgery was required in one case because of severe adhesion. The operation time was 75 min to 220 min (mean, 125 min), the blood loss was 50 ml to 420 ml (mean, 120 ml), and the postoperative hospital stay was 6 d to 12 d. Complications occurred in 4 cases. Pathological examination showed that 47 cases were of renal clear cell carcinoma, 5 of chromophobe carcinoma, and 1 of cystic renal cell carcinoma. Follow-up for 1 month to 5 years showed no tumor recurrence and metastasis.</p><p><b>CONCLUSION</b>Retroperitoneal laparoscopic radical nephrectomy is a safe and effective treatment for patients with stage T1 - 2N0M0 renal cell carcinoma.</p>

Mutation in the SLC37A4 gene of glycogen storage disease type Ib in 15 families of the mainland of China / 中华儿科杂志

<p><b>OBJECTIVE</b>Glycogen storage disease type Ib (GSDIb, MIM: 232220) is an autosomal recessive inborn error of metabolism caused by deficiency of the glucose-6-phosphate translocase. The clinical manifestations include symptoms and signs of both the typical GSDIa, including hepatomegaly, fasting hypoglycemia, lactic acidemia and hyperlipidemia, and the dysfunction of neutrophils of recurrent infection and neutropenia. More than 84 mutations have been identified since the discovery of the SLC37A4 gene as the disease causing gene. Up to date, 5 mutations in 4 Chinese patients were reported from Hong Kang and Taiwan. In order to see the spectrum of the SLC37A4 gene mutations and the correlation between genotype and phenotype in patients with GSDIb of the mainland of China, the authors investigated 17 GSDIb patients from 15 families in this study.</p><p><b>METHOD</b>Data of 17 patients from 12 provinces, 11 male and 6 female, aged 6 months to 35 years, were collected from the genetic clinics of Peking Union Medical College Hospital from Oct. 2006 to Mar. 2009. All of them were Han Chinese in ethnicity. Consanguineous status was confirmed in 2 unrelated patients. All patients were presented with hepatomegaly, fasting hypoglycemia, lactic acidemia, hyperlipidemia and neutropenia with variable frequency of infections. The full coding exons, their relevant exon-intron boundaries, and the 5'- and 3'-flanking regions of the SLC37A4 gene were amplified and directly sequenced. RT-PCR was performed to verify the effect of the 2 novel splicing mutations.</p><p><b>RESULT</b>A total of 11 mutations were identified in 15 families. Four mutations, p.Gly149Glu, p.Pro191Leu, p.Arg415X and c.1042_1043 del CT, were previously reported, and seven mutations, p. Leu23Arg, p.Gly115Arg, p.Gly281Val, p.Arg415Gly, c.784 + 1G > A, c.870 + 5G > A and c.1014_1120del107, were novel. The frequent mutations are p.Pro191Leu, p.Gly149Glu and c.870 + 5G > A, accounting for 37%, 15% and 11% of mutant alleles respectively. RT-PCR analysis of novel mutation c.784 + 1G > A confirmed the splicing of exon 5 of 159 bp, causing inframe deletion. While mutation c.870 + 5G > A was proved to cause exon 6, 86 bp, deletion causing frame-shift. Among 15 families, 12 genotypes were identified, including 3 with homozygous mutation and 9 with compound heterozygous mutations. Homozygous p.Pro191Leu mutation was the only genotype detected in more than 1 family and was found in 4 unrelated families, including 1 patient from consanguineous marriage.</p><p><b>CONCLUSION</b>A total of 11 SLC37A4 gene mutations were identified in 15 families of the mainland of China. The frequent mutations are p.Pro191Leu, p.Gly149Glu and c.870 + 5G > A. The number of Chinese SLC37A4 gene mutations was extended from 5 to 14.</p>

Structure and function of tonoplast Cation/H+ antiporters in plant: a review / 生物工程学报

Cation transporters play important roles in modulating the concentration of intracellular metal ions. The vacuole is an important storage organelle for many ions. Cation (Ca+)/H+ antiporters (CAXs) located at vacuolar membrane are mainly involved in the Ca2+ flux into the vacuole, and appear to be capable of transporting various divalent cations to some degree. Several CAX genes have been isolated and characterized from various plants in recent years. Four domains of plant CAXs have been identified: NRR regulates Ca2+ transport by a mechanism of N-terminal autoinhibition; Ca domain and C domain confer Ca2+ and Mn2+ specificity among CAX transporters, respectively; D domain plays a part in the regulation of cytosolic pH. AtCAXs identified in Arabidopsis thaliana are involved in the growth, development and stress adaption of plant. AtCAX3 is the mainly Ca2+/H+ transporter in response to salt stress; AtCAX2 and AtCAX4 participate in transportation and detoxicification of heavy metal ions (Cd2+, Zn2+, and Mn2+) in cells under heavy metal stress, and impact root/shoot Cd partitioning in plant. These suggest that CAX genes may be useful for nutritional enhancement of plants, and for increasing phytoremediation potential. Here, the classification, structure and function of CAXs in plants are reviewed.

Tetracycline and oxytetracycline resistance determinants detected in Bacillus cereus strains isolated from honey samples / Detección de determinantes de resistencia a tetraciclina y oxitetraciclina en cepas de Bacillus cereus aisladas de muestras de miel

The aim of this study was to investigate the presence of tetracycline and oxytetracycline resistance determinants in Bacillus cereus strains isolated from honey samples. Of a total of 77 isolates analyzed, 30 (39%) exhibited resistance to tetracyclines according to the results of a disk diffusion method. Resistant strains (n=30) were screened by PCR for the presence of the resistant determinants tetK, tetL, tetM, tetO, tetW, otrA and otrB and their MIC values for tetracycline, oxytetracycline and minocycline were assessed. According to the PCR results, 23 isolates (77%) presented at least one tetracycline or oxytetracycline resistance determinant. The tetK genotype was present in 10 isolates while the tetL, tetM, and otrA genotypes were present in 3, 2, and 5 isolates, respectively. In addition, 2 isolates of the tetK plus tetM genotype, 1 of the tetK plus tetL genotype, and 1 of the tetK plus otrA genotype were found. All isolates were tetW, tetO and otrB negatives. On the other hand, 7 isolates (23%) showed a tetracycline-resistant and/or minocyclineresistant phenotype (MIC) but did not carry any of the tet or otr determinants investigated in this study. This research has shown that B. cereus isolates from honey samples contain a variety of tetracycline and oxytetracycline resistance genes, including the tetK and tetL determinants which encode for efflux proteins, and tetM and otrA, which encode for ribosomal protection proteins. These findings indicate that strains isolated from honeys could represent a reservoir for tetracycline resistance genes. To our knowledge, this is the first report of tetracycline-resistant and oxytetracyclineresistant B. cereus strains carrying the tetK determinant, and also the first report of oxytetracycline-resistant and tetracycline- resistant Bacillus species carrying the otrA determinant.

El objetivo del presente estudio ha sido investigar la presencia de diversos determinantes de resistencia a tetraciclina y oxitetraciclina en las poblaciones de Bacillus cereus presentes en la miel. De un total de 77 aislamientos evaluados, 30 (39%) resultaron resistentes a tetraciclina y/o minociclina de acuerdo con los resultados de las pruebas de difusión en disco. Dentro del grupo que presentó un fenotipo resistente, se investigó la presencia de los determinantes tetK, tetL, tetM, tetO, tetW, otrA y otrB por PCR y se determinaron los valores de CIM para tetraciclina, oxitetraciclina y minociclina. De acuerdo con los resultados obtenidos por PCR, 23 aislamientos (77%) presentaron al menos un determinante de resistencia a tetraciclina o a oxitetraciclina; el genotipo tetK se encontró en 10 de esos aislamientos, mientras que los genotipos tetL, tetM y otrA se hallaron en 3, 2 y 5 aislamientos, respectivamente. Ningún aislamiento presentó los genotipos tetW, tetO ni otrB. Adicionalmente, se encontraron los genotipos tetK plus tetM (2 aislamientos); tetK plus tetL (1 aislamiento) y tetK plus otrA (1 aislamiento). Por otra parte, 7 cepas (23%) resultaron resistentes a tetraciclina, oxitetraciclina y/o minociclina por CIM, pero no presentaban ninguno de los determinantes tet u otr estudiados. Estos resultados indican la existencia de un alto porcentaje de cepas de B. cereus aisladas de miel con genes de resistencia a tetraciclina y oxitetraciclina, incluyendo los determinantes tetK, tetL, tetM y otrA. Este estudio constituye el primer registro de la presencia del determinante tetK de resistencia a tetraciclina en B. cereus, como así también la presencia del determinante otrA dentro del género Bacillus.

Isolation and molecular characterization of a cax gene from Capsella bursa-pastoris

A new cation exchangers (CAXs) gene was cloned and characterized from Capsella bursa-pastoris by rapid amplification of cDNA ends (RACE). The full-length cDNA sequence of cax from C. bursa-pastoris (designated as Cbcax51) was 1754 bp containing a 1398 bp open reading frame encoding a polypeptide of 466 amino-acid residues with a calculated molecular mass of 50.5 kDa and an isoelectric point of 5.69. The predicted CbCAX51 contained an IMP dehydrogenase/GMP reductase domain, two Na+/Ca2+ exchanger protein domains. Comparative and bioinformatics analyses revealed that CbCAX51 showed extensive homology with CAX from other plant species. The expression analysis by different treatments indicated that Cbcax51 could be activated by cold triggering and was related to the cold acclimation process, but its expression is regulated negatively by drought and not affected by ABA or salt.

The involvement of tetA and tetE tetracycline resistance genes in plasmid and chromosomal resistance of Aeromonas in Brazilian strains

This study analyzed the involvement of tetA and tetE genes in the tetracycline resistance of 16 strains of genus Aeromonas, isolated from clinical and food sources. Polymerase chain reactions revealed that 37.5 percent of the samples were positive for tetA, and also 37.5 percent were tetE positive. One isolate was positive for both genes. Only the isolate A. caviae 5.2 had its resistance associated to the presence of a plasmid, pSS2. The molecular characterization of pSS2 involved the construction of its restriction map and the determination of its size. The digestion of pSS2 with HindIII originated two fragments (A and B) that were cloned separately into the pUC18 vector. The tetA gene was shown to be located on the HindIII-A fragment by PCR. After transforming a tetracycline-sensitive strain with pSS2, the transformants expressed the resistance phenotype and harbored a plasmid whose size was identical to that of pSS2. The results confirmed the association between pSS2 and the tetracycline resistance phenotype, and suggest a feasible dissemination of tetA and tetE among strains of Aeromonas. This study suggests the spreading tetA and tetE genes in Aeromonas in Brazil and describes a resistance plasmid that probably contributes to the dissemination of the resistance.

Abnormal kinetics of erythrocyte sodium lithium countertransport in patients with diabetic nephropathy in Thailand.

BACKGROUND: In essential hypertension and diabetic nephropathy, sodium-lithium counter transport (Na/Li CT) is an inherited marker for metabolic influences of cardiovascular risk. The kinetics of Na/Li CT are modified by two types of thiol group in the membrane. In choline medium, the type 1 thiol reacts with N-ethtyl maleimide (NEM) to cause a decrease in Km and increase Vmax/Km ratio. However in the presence of external Na or Li both the type 1 or type 2 thiols react so that both Km and Vmax are reduced. Low Km of Na/Li CT has been previously reported to be a major abnormality in diabetic nephropathy (DN) and can be used to identify diabetic patients who are at high risk for DN. A recent study showed that the type 1 thiol protein controlling the Km of Na/Li CT was a 33-kD protein and the gene for this protein is going to be cloned. OBJECTIVE: The authors sought to identify Na/Li CT kinetic abnormalities in Type 2 diabetes in Thai patients. MATERIAL AND METHOD: Erythrocyte Na/Li CT kinetics and their modulation by thiol proteins were measured in erythrocytes from 22 patients with Type 2 diabetes and 42 normal control subjects. RESULTS: The kinetics of Na/Li CT in untreated erythrocytes were similar Thiol protein alkylation with NEM generally caused both Vmax and Km to fall, but caused Km to rise in erythrocytes of diabetic patients, whose native Km was low. Thus, abnormalities in the regulation of Na/Li CT by key thiol proteins were found in about one-third of subjects with Type 2 diabetes in Thailand. CONCLUSION: Membrane abnormalities may indicate a common pathway of pathological mechanism found in essential hypertension and diabetic nephropathy and may be used as a phenotype for further genetic studies of this transporter.

Analysis of the interaction between human kidney anion exchanger 1 and kanadaptin using yeast two-hybrid systems

Kidney anion exchanger adaptor protein (Kanadaptin) is a protein which interacts with the cytoplasmic N-terminal domain of kidney anion exchanger 1 (kAE1) and was first detected in mice using the yeast two-hybrid system and was also found to co-localize with kAE1 in rabbit a-intercalated cells. Impaired trafficking of human kAE1 can result in the kidney disease-distal renal tubular acidosis (dRTA), and defective interaction between human kAE1 and kanadaptin may cause this trafficking impairment and be the basis for dRTA pathogenesis. However, it is unknown whether kAE1 can really interact with kanadaptin in humans. We have thus investigated the interaction between human kAE1 and human kanadaptin by using both Gal4 and LexA yeast two-hybrid systems. It was found that co-expression of Gal4DBD fused to the cytoplasmic N-terminal domain of kAE1 and Gal4AD fused to kanadaptin could not activate the transcription of the ADE2, HIS3 and lacZ reporters in the Gal4 system. A similar result was obtained for the interaction between B42AD fused to the cytoplasmic N-terminal domain of kAE1 and LexA fused to kanadaptin in activation of lacZ transcription in the LexA system. The absence of interaction between the fusion proteins in both yeast two-hybrid systems raises the possibility that kAE1 may not interact with kanadaptin in human cells. Considerably different structures of both kAE1 and kanadaptin in mice and humans may lead to different binding properties of the proteins in these two species.

Clinicopathologic and immunohistochemical study of pulmonary epithelioid hemangioendothelioma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinical and pathological characteristics of pulmonary epithelioid hemangioendothelioma.</p><p><b>METHODS</b>Four cases of pulmonary epithelioid hemangioendothelioma were studied by histopathologic and immunohistochemical examination of lung biopsy specimens.</p><p><b>RESULTS</b>There were 3 female and 1 male, age 28 to 40 years. Clinically the tumor presented as multiple bilateral small nodules in the lung. Histologically, crown-like clusters of epithelioid tumor cells were obtained which filled in the alveoli locating at the periphery of the tumor nodules, while the central part of the nodules contained myxoid to hyaline matrix. The overall architecture of the lung was still preserved. Additionally, intracytoplasmic vacuoles were seen in tumor cells within which red blood cells were sometimes identified. Tumor cells generally lacked pleomorphism, mitotic activity and necrosis. They were immunohistochemically positive for CD31 and CD34. AE1/AE3 staining was positive in some cases.</p><p><b>CONCLUSIONS</b>Pulmonary epithelioid hemangioendothelioma often occurs in a middle-aged woman and represents a distinct clinical pathological entity.</p>

Lithium clearance and sodium lithium counter transport in insulin dependent diabetes mellitus

Our study was conducted on 24 insulin dependent diabetic patients in comparison to 10 normal healthy control subjects. the group of the patients were classified according to the results obtained into 3 groups. The list was eight diabetic patients without nephropathy, the second was eight diabetic patients with nephropathy and the third was eight diabetic hypertensive patients, Both diabetics and control were subjected to full clinical examination and laboratory investigations [plasma lithium, lithium clearance, fractional Iithium clearance [FcLi] and sodium-lithium counter transport [Na [+] /Li [+] CT]. The results showed mild elevation of serum sodium in IDDM groups compared to control. FcLi showed a highly reduction in IDDM groups compared to control group. Na [+] /Li [+] CT showed a statistically significant elevation in IDDM groups compared to control group

Effect of glandular kallikrein on distal bicarbonate transport. Role of basolateral Cl-/HCO3- exchanger and vacuolar H(+)-ATPase

The luminal membrane of collecting duct cells, specially the intercalated cells, is normally exposed to active kallikrein. This is due to the specific localization of renal kallikrein in the connecting tubule cells. We have previously reported inhibition of distal bicarbonate secretion by renal kallikrein. The present study was performed to evaluate the participation of basolateral Cl-/HCO3- exchanger and luminal H(+)-ATPase activity of cortical collecting duct segments (CCD) in the mechanism involved in the inhibition of bicarbonate secretion induced by the enzyme. The effect of orthograde injections of 1 microgram/ml (250 U/6.3 mg) pig pancreatic kallikrein, in the absence and presence of 1 mM DIDS (stilbene-disulfonic acid) in the renal tubule system, was evaluated. Urine fractions were collected after two-minutes stop-flow. Changes in the urine fraction (Fr) related to those in free-flow urine samples (Ff) were related to the respective polyfructosan (Inutest) ratio. Renal kallikrein activity (Fr:Ff kallikrein/Fr:Ff polyfructosan) increased significantly in the first 120 microliters urine fraction collected after glandular 1 microgram/ml kallikrein, P < 0.05, (first stop-flow) and after glandular 1 microgram/ml kallikrein plus 1 mM. DIDS P < 0.05 (second stop flow). Bicarbonate secretion rate (Fr:Ff HCO3-/Fr:Ff polyfructosan) of collecting ducts was significantly reduced in the first 120 microliters urine fraction collected, related to control, during the first and second stop-flow periods. No difference was shown in bicarbonate excretion between the first 120 microliters urine fractions collected after administration of glandular kallikrein and glandular kallikrein plus DIDS. To measure H(+)-ATPase activity, rat microdissected cortical collector tubules (CCD) were incubated in the presence of increasing glandular kallikrein doses (A: 93, B: 187 and C: 375 mU/200 microL) in the presence of ouabain (4 microM) and omeprazole (100 microM) to inhibit Na(+)-K(+)-ATPase and H(+)-K(+)-ATPase, respectively. In CCD, bafilomycin-sensitive H(+)-ATPase activity (pmol/mm/min) after increasing kallikrein doses did not differ significantly from control...

Ca2+ transport and oxidative damage of mitochondria

1. Mitochondria from a wide range of sources have the ability to accumulate Ca2+ down their electrochemical gradient mediated by a uniport mechanism. 2. Ca2+ efflux occurs via two separate pathways: a Na+/Ca2+ exchanger that predominates in mitochondria from excitable tissues and a Na(+)-independent pathway that predominates in mitochondria from non-excitable tissues. 3. The kinetic characteristics of these calcium influx-efflux pathways appear to be incompatible with any role for mitochondria as cytosolic Ca2+ buffers, under resting normal physiological conditions. Instead, the biological role of this Ca(2+)-transporting system seems to be the regulation of matrix Ca2+ in a range that permits the regulation of three intramitochondrial Ca(2+)-dependent dehydrogenases which catalyze rate-limiting reactions of the Krebs cycle. 4. Under conditions in which a high cytosolic Ca2+ concentration is sustained, the matrix Ca2+ concentration may attain levels that lead to impairment of mitochondrial functions such as inhibition of oxidative phosphorylation and increase in inner membrane permeability. 5. Accumulation of Ca2+ by mitochondria under conditions of oxidative stress induces an increase in inner membrane permeability by a mechanism that appears to be mediated by protein polymerization due to thiol cross-linking