Objective To explore the potential of converting the chloramphenicol resistant clinical E. Coli isolates to chloramphenicol sensitive ones by employing external guide sequence(EGS) technique in vitro. Methods Recombinant plasmids with EGScat 1+2 and tetracycline resistant gene, named PAlterl EGScat1+2, was constructed. Routine Cacl 2 method was used to introduce recombinant plasmid into the chloramphenicol resistant clinical isolates E. Coli 4758. Colony PCR was used to test and A600 was used to detect growth rates in liquid and solid culture of various concentrations of chloramphenicol. Results The chloramphenicol resistant clinical isolates E. Coli 4758 grew well in chloramphenicol(35 ?g/ml, 70 ?g/ml, 105 ?g/ml, 170 ?g/ml) plates whereas the transformants tE4758 with PAlter1 EGScat 1+2 failed to grow in these concentrations, which indicated its resistance to the chloramphenicol was reversed. Conclusions EGS molecules are able to convert the drug resistance in clinical E. Coli isolates in vitro.

OBJECTIVETo demonstrate the effects and mechanisms of Huangkui capsule (HKC) on renal fibrosis in rats with diabetic nephropathy (DN).

METHODRats were randomly divided into 5 groups, the sham-operated group (Sham group, n = 5), the vehicle-given group (Vehicle group, n = 7), the low dose of HKC-treated group (L-HKC group, n = 7), the high dose of HKC-treated group (H-HKC group, n = 7) and the lipoic acid (LA)-treated group (LA group, n = 7). DN models were induced by intraperitoneal injection of streptozotocin (STZ,35 mg x kg(-1)) twice and unilateral nephrectomy. After models were successfully established, the rats in HKC and LA groups were daily administrated with HKC suspensions (0.75, 2 g x kg(-1)) or LA suspensions (60 mg x kg(-1)) respectively, and at the same time, the rats in Vehicle group were daily administrated with distilled water (2 mL) for 8 weeks. All rats were sacrificed at the end of week 8 to collect blood and renal tissues. UAlb, renal function, renal fibrotic morphologic characteristics, as well as oxidative stress (OS)-related markers, the protein expressions of the key signaling molecules in p38 mitogen-activated protein kinase (p38MAPK) signaling pathway, fibrogenic cytokines and inflammatory factors were examined respectively.

RESULTHKC, similar to LA, improved the general state of health, body weight, UAlb, BUN, UA and Alb in DN model rats. Of note, renal fibrosis was ameliorated in HKC groups,especially in H-HKC group which was better than that in LA group. In addition, HKC not only improved the main indexes of OS in the kidney like LA, but also down-regulated the protein expressions of phosphorylated-p38MAPK (p-p38MAPK), transforming growth factor (TGF)-β1 and tumor necrosis factor(TNF)-α in the kidney, whereas, LA only decreased the protein expression of TNF-α in the kidney in DN model rats.

CONCLUSIONHKC, similar to LA, has the actions of anti-OS in vivo. Moreover, HKC could attenuate renal fibrosis by suppressing the activation of p38MAPK signaling pathway and the protein expressions of fibrogenic cytokines and inflammatory factors in the kidney in DN model rats, which is different from LA.

Abelmoschus ; chemistry ; Animals ; Capsules ; Diabetic Nephropathies ; drug therapy ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; Kidney ; drug effects ; pathology ; MAP Kinase Signaling System ; drug effects ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

In the development of diabetic nephropathy (DN), reactive oxygen specie (ROS) over much in vivo leads to oxidative stress(OS)-related renal injuries, which are characterized by the structural and functional changes in glomerular and renal tubular cells in morphology. The regulative approaches of OS involve the several signaling pathways, in which, both p38 mitogen-activated protein kinase (MAPK) signaling pathway and adenosine monophosphate-activated protein kinase (AMPK) signaling pathway play the important roles as the target of anti-oxidants. The interventional actions of Chinese herbal compound prescriptions and the extracts of single Chinese herbal medicine (CHM) on OS in the kidney in DN include regulating the balance between ROS and antioxidants, reducing the production of AGEs, inhibiting the expression of growth factors and intervening the activity of signaling pathways.
Animals ; Diabetic Nephropathies ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Kidney ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Signal Transduction ; drug effects ; Treatment Outcome

Objective · To investigate the impacts of hepatitis B virus (HBV) infection on the metabolomic phenotype of HepG2 human hepatoma cells.Methods · With gas chromatography-mass spectrometry (GC-MS), metabolite composition of HepG2 and HepG2.2.15 cells (derived from HepG2 cells transfected with a plasmid containing HBV) were analysed. Results · GC-MS analysis mainly found 34 metabolites in both HepG2 and HepG2.2.15 cells,including glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), serine (Ser), threonine (Thr), methionine (Met), cysteine (Cys), cystine, aspartic acid (Asp), glutamic acid (Glu), pyroglutamic acid, phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), hypoxanthine, uracil,myo-inositol, lactic acid, succinic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid, urea, cholesterol, etc. These metabolites were involved in multiple metabolic pathways including glycolysis and metabolism of fatty acids, amino acids, purines and pyrimidines. Compared with HepG2 cells,HepG2.2.15 cells had significantly higher levels in lactic acid, linolenic acid, Ala and Cys, but lower levels in Leu, Ile, Val, Phe, Met, Trp, Pro, Tyr, myoinositol and uracil. Conclusion · HBV infection dysregulates the metabolism of amino acids and fatty acids in hepatocytes. GC-MS analysis provides complimentary information about HBV-induced metabolic changes of host cells.

OBJECTIVETo determine the prevalence and types of GJB2 mutations and to investigate the genetic mechanism in Chinese autosomal recessive deafness.

METHODSThe subjects were four Chinese pedigrees (39 individuals) and 50 normal adults. GJB2 was amplified by PCR. The products were digested with restriction enzyme Apa I, then sequenced.

RESULTSHomozygous deletion C at position 232-235 of GJB2 (235delC),which resulted in frameshift mutation, was found in four affected individuals of two pedigrees; the compound heterozygous deletions (235delC/232G to A) were found in two affected individuals in one pedigree. One carrier with 235delC was found in normal controls (1% allele). Two kinds of polymorphisms 79G to A(V27I) and 3 41A to G(E114G) were found in both affected and normal controls. The frequencies of allele for 79G to A and 341A to G in normal controls were 30%, 21%, respectively.

CONCLUSION235delC mutation of GJB2 was related with Chinese autosomal recessive deafness, and the 232G to A(Ala78Thr) missense mutation was found to be a novel mutation.

Base Sequence ; China ; Connexin 26 ; Connexins ; genetics ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Deafness ; genetics ; Family Health ; Female ; Humans ; Male ; Mutation ; Mutation, Missense ; Pedigree ; Sequence Deletion

OBJECTIVETo investigate the expression of four hepatocellular cancer antigen (HCA) gene mRNA in hepatocellular carcinoma.

METHODSThe expression of HCA90, HCA519, HCA520, HCA587 mRNA was detected using RT-PCR in HCC tissues and the corresponding adjacent non-HCC tissues from 46 HCC patients, cirrhosis tissues from 10 samples and normal liver tissues from 10 samples. The relationship between positive expression rate of HCA gene and clinical and lab data was evaluated.

RESULTSOf 46 HCC tissues, HCA90, HCA519, HCA520 and HCA587 mRNA were detectable in 65.2%, 76.1%, 45.7% and 32.6%, respectively. At least one HCA gene mRNA was positive in 82.6% of HCC tissues. Only weak expression of HCA519 could be detectable in 6.5% of the corresponding adjacent non-HCC tissues. None of 10 samples of cirrhosis and normal liver tissues expressed any HCA gene mRNA. No correlation was found between the expression of HCA and clinical date such as age, sex, tumor size, tumor differentiation, serum alpha-fetoprotein level and hepatitis B virus infection or hepatitis C virus infection (P > 0.05). However, in some patients with normal serum alpha-fetoprotein (< 25 ng/L), specific expression of HCA genes was observed.

CONCLUSIONHCA gene mRNA is expressed with a high percentage and specificity in hepatocellular carcinomas and their products are new potential promising targets for immunotherapy of HCC.

Antigens, Neoplasm ; genetics ; Carcinoma, Hepatocellular ; genetics ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Liver ; metabolism ; Liver Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVESTo evaluate the efficacy of intrameatal application of low dosage alprostadil (PGE1) cream (300 mcg) for the treatment of erectile dysfunction (ED).

METHODSA total of 43 ED patients were selected in the study based on the inclusion criteria. All of the patients signed informed consent forms and entered a 4-week open-label clinical study. A dosage of 300 mg PGE1 in 75 mg cream was applied intrameatally.

RESULTSThe results showed that the primary efficacy (IIEF Q3 + Q4) reached 70.73% after application of the cream. The successful intercourse rate was 86.41%. Based on the GAQ (global assessment Question); 73.17% of the patients were satisfied with their sexual life. At the same time, all of the secondary criteria supported the primary efficacy results. Two patients withdrew during the study period. Six patients (14.63%) had urethral pain or penile redness, which were mostly mild and transient.

CONCLUSIONSWith intrameatal low dosage (300 mcg PGE1) of the PGE1 cream can achieve an equivalent efficacy as that with the full dosage.

Adult ; Aged ; Alprostadil ; administration & dosage ; therapeutic use ; Erectile Dysfunction ; drug therapy ; Humans ; Male ; Middle Aged ; Penile Erection ; drug effects ; Treatment Outcome ; Vasodilator Agents ; administration & dosage ; therapeutic use

It is reported, in the process of diabetic nephropathy (DN), inflammatory-related p38 mitogen-activated protein kinase (MAPK) signaling pathway has a close relationship with renal injury. On the one hand,many factors in the upstream including hyperglycemia, abnormal hemodynamics, oxidative stress, and pro-inflammatory cytokines could activate p38MAPK signaling pathway. On the other hand,the activated p38MAPK signaling pathway could lead to renal damage via activating inflammatory cells, inducing the expression of inflammatory mediators, and intervening cytokines production. CHM could intervene p38MAPK signaling pathway through multi-ways, including inhibiting inflammatory cytokines expression, regulating phosphorylated p38MAPK (p-p38MAPK) expression, and reducing fibrogenic factors expression.
Diabetic Nephropathies ; drug therapy ; enzymology ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Inflammation ; drug therapy ; metabolism ; Inflammation Mediators ; pharmacology ; Kidney ; drug effects ; enzymology ; metabolism ; Signal Transduction ; drug effects ; p38 Mitogen-Activated Protein Kinases ; metabolism

BACKGROUNDHuman amniotic epithelial cells (HAECs), which have characteristics of both embryonic and pluripotent stem cells, are therefore a candidate in cell therapy without creating legal or ethical problems. In the present study, we aimed to investigate the effects of intracerebroventricular transplantation of HAECs on doubly transgenic mice of Alzheimer's disease (AD) coexpressing presenilin-1 (PS1) and mutant Sweden amyloid precursor protein (APPswe) genes.

METHODSThe offspring mice genotypes were detected using PCR identification of APPswe and PS1 gene. The doubly transgenic (TG) mice (n = 20) and wild-type (WT) mice (n = 20) were randomly divided into two groups respectively: the transplantation group treated with HAECs and the control group with phosphate buffered saline. Six radial arm water maze test was used to assess the spatial memory in the TG and WT mice. Amyloid plaques and neurofibrillary tangles were analyzed using congo red and acid-silver methenamine staining respectively. Immunofluorescence cytochemistry was used to track the survival of HAECs. Immunohistochemistry was used to determine the expression of octamer-binding protein 4 (Oct-4) and Nanog in the HAECs. High performance liquid chromatography was used to measure acetylcholine in hippocampus. The density of cholinergic neurons in basal forebrain and nerve fibers in hippocampus was measured using acetylcholinesterase staining.

RESULTSAmyloid deposition occurred in hippocampus and frontal cortex in the double TG mice aged 8 months, but not in WT mice. The results also showed that transplanted HAECs can survive for at least 8 weeks and migrate to the third ventricle without immune rejection. The graft HAECs can also express the specific marker Oct-4 and Nanog of stem cell. Compared with the control group, transplantation of HAECs can not only significantly improve the spatial memory of the TG mice, but also increase acetylcholine concentration and the number of hippocampal cholinergic neurites.

CONCLUSIONSThese results demonstrate that intracerebroventricular transplantation of HAECs can improve the spatial memory of the double TG mice. The higher content of acetylcholine in hippocampus released by more survived cholinergic neurites is one of the causes of this improvement.

Acetylcholine ; metabolism ; Alzheimer Disease ; genetics ; metabolism ; therapy ; Amnion ; cytology ; Amyloid beta-Protein Precursor ; genetics ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Epithelial Cells ; cytology ; transplantation ; Genotype ; Hippocampus ; metabolism ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Memory Disorders ; genetics ; metabolism ; therapy ; Mice ; Mice, Transgenic ; Nanog Homeobox Protein ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Polymerase Chain Reaction ; Presenilin-1 ; genetics ; metabolism

OBJECTIVETo evaluate the diagnostic value of serum angiopoietin-2 (Ang-2) level in pancreatic cancer patients.

METHODSSerum Ang-2 level was measured by enzyme-linked immunosorbent assays (ELISA) in samples from 116 patients with pancreatic cancer, 50 patients with chronic pancreatitis, and 50 normal control subjects.

RESULTSThe serum Ang-2 level in patients with pancreatic cancer [(1539.0 ± 449.3) ng/L] was significantly higher than those in patients with pancreatitis [(1044.6 ± 246.1) ng/L, P < 0.01] and normal control subjects [(1075.6 ± 228.2) ng/L, P < 0.01]. There was no significant difference of serum Ang-2 levels between patients with pancreatitis and normal controls. Pancreatic caner patients with lymph node metastasis had a significantly higher serum Ang-2 level [(1890.1 ± 354.9) ng/L] than those without metastasis [(1212.1 ± 224.2) ng/L, P < 0.01]. The area under ROC curve of serum Ang-2 level for diagnosis of pancreatic cancer was 0.819.

CONCLUSIONThe serum Ang-2 level can be a useful indicator for diagnosis of pancreatic cancer.

Adult ; Aged ; Angiopoietin-2 ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Pancreatic Neoplasms ; blood ; diagnosis ; pathology ; Pancreatitis ; blood ; ROC Curve ; Retrospective Studies