Carrier Proteins ; metabolism ; physiology ; HIV-1 ; physiology ; Models, Biological ; Proteins ; metabolism ; physiology ; Retroviridae Infections ; virology

Objective To investigate the distribution of surfactant protein-C( SP-C) gene single nu-cleotide polymorphisms and to study the association between the SP-C gene polymorphisms and neonatal respiratory distress syndrome( NRDS) in infants. Methods Fifty-one infants with NRDS( NRDS group) and 51 infants without RDS( control group) were selected. PCR gene analysis and polymerase chain reaction were used to establish the genotype and allele frequencies of SP-C exon 4(T138N) and exon 5(S186N),SP-C exon 4 and 5 for the mutation,and then the association between the polymorphisms and NRDS was analyzed. Results SP-C gene mutations were not found in exon 4 and 5. In the Mongol nationality of the Inner Mon-golia region,SP-C exon 4(T138N) genotypes could check out three genotypes:namely AA,AC and CC. The frequencies of allele A and allele C of SP-C exon 4(T138N) were not statistically different between NRDS group and control group(χ2 =0. 454,P=0. 797). In the Mongol nationality,SP-C exon 5(S186N) genotypes could check out three genotypes:namely AA,AG and GG. The frequencies of allele A and allele G of SP-C exon 5(S186N) were not statistically different between NRDS group and control group(χ2 =0. 493,P =0. 782). Conclusion SP-C exon 4(T138N) and exon 5(S186N) gene polymorphism in Inner Mongolia newborns displays no significant correlation with sex,birth weight or gestational age. SP-C gene mutations are not found in exon 4 and 5. SP-C gene exon 4(T138N) and exon 5(S186N) polymorphisms are not found to be associated with NRDS in Mongol nationality of the Inner Mongolia.

DNA Fingerprinting ; DNA, Mitochondrial ; Homicide ; Humans ; Mitochondria

OBJECTIVETo investigate the expression and significance of Cyclin D1 in oral squamous cell ma (OSCC).

METHODSA immunohistochemistry method, Envosion, was employed to test the manifesting Cyclin D1 in pathological slices of 50 OSCC cases and 10 normal cases, and the results was treated with statistical lysis.

RESULTSIn 50 OSCC cases, Cyclin D1 mainly manifested in karyon, and a little in cytoplasm. manifesting rates of Cyclin D1 in the samples was 80.0%, which was significantly higher than the manifesting of 20.0% in normal oral mucous membrane (P < 0.01). The manifestation of Cyclin D1 was correlated with rent pathological grades, clinical phases and lymph node metastasis (P < 0.05).

CONCLUSIONThe abnormal tation of Cyclin D1 is closely related with the occurrence and development of OSCC. Therefore, it can subsidiary index for OSCC treatment and prognosis.

Carcinoma, Squamous Cell ; Cyclin D1 ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Mucosa ; Mouth Neoplasms ; Prognosis

OBJECTIVETo investigate the effect of resistin overexpression on 3T3-L1 adipocyte lipid and glucose metabolism.

METHODSExpression vector for rat resistin gene was constructed and transfected into 3T3-L1 adipocytes. Cell differentiation and lipid accumulation was determined by Oil Red O staining. Differentiation marker genes (pref-1, C/EBPalpha, FAS) and glucose transporter 4 (GLUT4) gene mRNA expressions were evaluated by reverse transcription-PCR (RT-PCR). Triglyceride (TG) and free fatty acids (FFAs) in adipocytes were measured by colorimetric kit.

RESULTS(1) In resistin-overexpressed adipocytes, the lipid droplets presented at the second day which was earlier than the control cells. (2) The expression of C/EBPalpha and FAS genes in resistin-overexpressed adipocytes were up-regulated and the pref-1 was down-regulated compared with that of the control cells. (3) In resistin-overexpressed adipocytes, cellular TG and FFAs levels were significantly increased (P<0.05). (4) There was no difference in the expression of GLUT4 gene between 3T3-L1 adipocytes and resistin-overexpressed adipocytes (P> 0.05).

CONCLUSIONOverexpression of resistin can affect 3T3-L1 adipocyte lipid metabolism and thereby result in obesity and insulin resistance, but have no effect on GLUT4 gene expression.

3T3-L1 Cells ; Adipocytes ; metabolism ; Animals ; Cell Differentiation ; genetics ; Fatty Acids, Nonesterified ; metabolism ; Gene Expression ; Glucose Transporter Type 4 ; genetics ; Lipid Metabolism ; genetics ; Mice ; Rats ; Resistin ; genetics ; metabolism ; Triglycerides ; metabolism

Objective To analyze the surveillance results and grasp the situation of Keshan disease in Wudalianchi city Heilongjiang province.Methods In 2009,Kaifa village was selected as the surveillance point in Wudalianchi city,total resident population were monitored by routine clinical examination and 12-lead electrocardiogram(ECG) tracing.Suspected cases with Keshan disease were taken chest X-ray,and Keshan disease was diagnosed based on Keshan Disease Diagnostic Criteria (WS/T 210-2011).Results A total of 795 people were investigated,including 397 males and 398 females.Eighteen people were found to be the patients with Keshan disease,of which 13 cases were latent Keshan patients,5 cases were chronic Keshan patients.The overall detection rate was 2.27％,aged 24 to 83 years old.There was no acute type and subacute type of Keshan disease in the surveillance point.Twenty nine cases of abnormal ECG were detected,the detection rate was 3.65％ (29/795),of which the 18 patients with Keshan disease were all had abnormal ECGs,mainly taken the form of ST-T changes and completely right bundle branch blocked.Six cases of male patients with Keshan disease were detected,the detection rate was 1.52％ (6/397); 12 cases of female patients with Keshan disease were detected,the detection rate was 3.01％ (12/398).Conclusions There is still potential and chronic Keshan disease cases in Wudalianchi city.We must keep on the monitoring on Keshan disease,master the dynamical changes of the disease conditions,and carry out the targeted prevention and control of Keshan disease.

BACKGROUNDMonilethrix is an autosomal dominant hair disorder characterized clinically by alopecia and follicular papules. In this study, we collected a Han monilethrix family to detect the mutations in patients and investigated the correlation between the genotype and phenotype of monilethrix.

METHODSIn this study, we identified a Chinese family with monilethrix through light microscopic and scanning electron microscopic (SEM) examination. Genomic DNA from peripheral blood samples was prepared. DNA samples from controls and monilethrix patients were subject to polymerase chain reaction (PCR) amplification. Two pairs of primers were used to amplify the seventh exon of KRT86. Mutation screening of the PCR products was detected using direct sequencing.

RESULTSLight microscopic examination showed a regular alternate enlargement and narrow area. SEM examination showed that part of the cuticle of the nodules shed and disappeared gradually in the narrow area with granular protrusions on the surface similar to the erosion-like structure. Parallel longitudinal ridge and groovepattern appeared, and the ridges varied in width, like dead wood. A heterozygous transversion mutation c.1204G > A (p.E402K) in the seventh exon of KRT86 was identified in both patients.

CONCLUSIONSThe mutation of extron 7 of KRT86 identified plays a major role in the pathogenesis of this pedigree with monilethrix, and is a mutation hot spot of KRT86. Further research is needed to explore the relationship between the phenotype and the mutation of the type II hair keratin gene KRT86 of monilethrix.

Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Humans ; Keratins, Hair-Specific ; genetics ; Keratins, Type II ; genetics ; Microscopy, Electrochemical, Scanning ; Monilethrix ; etiology ; genetics ; pathology ; Mutation

OBJECTIVETo identify the self-preparation monoclonal antibody which target to clenbuterol, and set up the standard curve to clenbuterol (CL) detection.

METHODSThe affinity constants and activity of the monoclonal antibody which target to CL were determined by ELISA. ELISA was also used to confirm whether the monoclonal antibody had any across-reaction with BSA and CL analogues. The rat ascites which contains the monoclonal antibody target to CL was purified by (NH4)2SO4 salt-out method and further by affinity column. At last, the CL detection standard curve which based on indirect competition ELISA was established.

RESULTSThe ELISA experiment showed that the antibody titer was 10(6) and the monoclonal antibody affinity constants was 2.90 x 10(10) L/mol. The result of the indirect competition ELISA confirmed that the monoclonal antibody had no cross-reaction with BSA and a few kind of CL analogue. CL detection standard curve based on indirect competition ELISA was established, which R2 was 0.9812, and the lowest detectable limit was 1.0 ng/ml.

CONCLUSIONThe standard curve based on indirectly competitioning ELISA was established. The self-preparation monoclonal antibody which target to CL has high affinity and high specific to CL, which had established the foundation to the advanced development of the CL immune test paper and CL ELISA kit.

Animals ; Antibodies, Monoclonal ; chemistry ; isolation & purification ; Antibody Affinity ; Clenbuterol ; immunology ; Cross Reactions ; Enzyme-Linked Immunosorbent Assay ; Limit of Detection ; Rats

OBJECTIVETo explore feasible and reliable methods for estnbolishment and of acute lung injury model in animal models.

METHODSTwenty-four healthy adult mongrel dogs with oleic acid-induced acute lung injury were evaluated for early-stage physiological response to acute lung injury using a balloon catheter system for measuring esophageal and gastric pressure.

RESULTSIn canine models of early-stage oleic acid-induced acute lung injury that sustained spontaneous breathing, in terms of respiratory mechanics, some parameters obviously increased including the respiratory rate (RR), minute ventilation (VE), peak inspiratory volume (Vinsp, peak), mean inspiratory volume (VT/Ti), inspiratory airway resistance (Raw, insp) (P<0.001 for all the parameters), with also significantly increased peak transdiaphragmatic pressure (Pdi, peak, P=0.0185). The tidal volume (VT) and dynamic lung compliance (CL,dyn), however, were significantly decreased (P<0.001), and significant variation occurred in the ratio of inspiratory time to duration of one breath (Ti/Ttot, P=0.163). In terms of gas exchange, the pH, PaO(2), SaO(2), PaO(2)/FiO(2), and end tidal partial pressure of carbon dioxide (PETCO(2)) all evidently declined (P<0.001), but PaCO(2) and ratio of alveolar dead space to tidal volume [VD(alv)/VT] increased significantly (P<0.001).

CONCLUSIONApplication of balloon catheter system for measuring esophageal and gastric pressures allows objective evaluation of the various physiological responses in early stage of acute lung injury.

Acute Lung Injury ; physiopathology ; therapy ; Animals ; Catheterization ; Disease Models, Animal ; Dogs ; Esophagus ; Feasibility Studies ; Female ; Male ; Pressure ; Reproducibility of Results ; Respiratory Mechanics ; physiology ; Stomach ; Time Factors

Loop-mediated isothermal amplification (LAMP) assay is a novel method of gene amplification with high specificity, sensitivity and rapidity, which can be applied for disease diagnosis in shrimp aquaculture. The method is performed under isothermal conditions with a set of four specially designed primers that recognize six distinct sequences of the target. In the present study, according to the conservative regions of non-structural protein gene NS1, a set of four specific primers were designed, and a rapid detection of IHHNV was established by LAMP assay. The parameters of reaction time and temperature were optimized, and its specificity and sensitivity were assessed. The reactions were carried out at 60 degrees C, 62 degrees C, 63 degrees C, 64 degrees C, 65 degrees C, 66 degrees C, 67 degrees C, 68 degrees C for different time (0 min; 15 min; 30 min; 45 min; 60 min; 75 min). A plasmid pMDIHHNV carrying target sequence of LAMP detection was constructed. Ten-fold serially diluted pMDIHHNV (10(7)-10(0)copies/microL) was used as template for LAMP assay to investigate the detection limit. To determine the specificity, LAMP assays were carried out with DNA templates from other pathogens (White spot syndrome virus; WSSV, Taura Syndrome Virus; TSV, Aeromonas. hydrophila, V. alginolyticus, Vibrio. parahaemolytious, Escherichia. coli). The results showed the optimized LAMP assay for the rapid detection of IHHNV was performed at 65 degrees C for 60 min. The LAMP assay had an unequivocal detection limit of 100 copies/microL, and it was 1,000 times lower than that of PCR. The nucleic acids of other pathogens were not amplified by this LAMP system with the specific primers, which showed a good specificity. The resulting amplicons were detected using visual observation after the addition of SYBR Green I and gel electrophoresis. We investigated the efficacy of UNG (uracil-N-glycosylase) and dUTP in avoiding carry-over contamination in the LAMP assay procedure and explored its effect on the amplification efficiency. Products of LAMP with dUTP adding could be lysed by UNG to avoid LAMP products carry-over contamination effectively. The LAMP assay could be finished within an hour, requiring only a regular laboratory water bath or heat block for reaction and the result could easily be detected using visual observation. Clinically suspected IHHNV-infected shrimp samples were detected by both LAMP and PCR assay, and the result indicated that IHHNV was detected rapidly by LAMP instead of by PCR.
Animals ; DNA Primers ; genetics ; Densovirinae ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Penaeidae ; virology ; Viral Proteins ; genetics