Objective To investigate the relationship between single nucleotide polymorphisms of surfactant protein C(SP-C)gene and respiratory distress syndrome(RDS)in the Han nationality newborns in Inner Mongolia and whether there is a mutation occurs on SP-C gene exon 4 and 5.Methods One hundred newborns with RDS(case group)and 100 newborns without RDS(control group)were selected.PCR gene analysis was used to establish the genotype and allele frequencies of exon 4 (T138N)and 5 (S186N)on SP-C.Results In the Han nationality newborns of Inner Mongolia region,there was no mutation on SP-C gene exon 4 and 5.Exon 4(T138N)on SP-C could be checked out three genotypes:namely AA,AC and CC.The genetic polymorphisms of exon 4 on SP-C were not statistically different between the case group and the control group(χ2 ﹦0.744,P ﹦0.689).Besides,exon 5(S186N)on SP-C could also be checked out three genotypes:namely AA,AG and GG.The genetic polymorphisms of exon 5 on SP-C were also not statistically different between the case group and the control group(χ2 ﹦0.770,P ﹦0.681 ).Conclusion There is no mutation on SP-C gene exon 4 and 5.The genetic polymorphism of exon 4 and 5 on SP-C displays no signifi-cant correlation with RDS of the Han nationality newborns in Inner Mongolia.

Objective To explore the effect of carvedilol on ACE2 gene and protein expression in chronic heart failure rats after myocardial infarction.Methods The heart failure model was induced by acute myocardial infarc- tion (AMI) through ligating the left anterior descending coronary artery.One month after operation,rats were randomized to receive placebo or carvedilol 2 mg/(kg?d),by gavage.Sham-operated rats were used as the control group.Hemodynamies,body mass and left ventrieular mass index,plasma and myocardial level of angiotensin Ⅱ were determined.ACE2 gene and protein expression was assessed by using RT-PCR and Western Blot.Results The mortality of placebo and Carvedilol groups were 20%,compared with 0% in sham operated rats.Carvedilol significantly improved LVEDP,LVSP,+dp/dt_(max) and-dp/dt_(min) in CHF rats but all the hemodynamics data were still inferior than that of controls.Plasma and myocardial angiotensin Ⅱ level were increased significantly in CHF placebo rats than those of control rats (plasma Ang Ⅱ:CHF:194?19 vs controls:132?15 ng/L,myocardium Ang Ⅱ:CHF:6.7?0.4 vs control:3.8?0.3 ng/g,P

Objective To study the correlation between the surfactant protein C ( SP-C) gene mutation in exon 5 area and respiratory distress syndrome(RDS) in premature infants. Methods From January 2013 to January 2015, nonconsanguineous premature infants [28 weeks ≤gestational age(GA)< 37 weeks] of Han ethnicity with RDS admitted to our hospital were selected as the RDS group, and nonconsanguineous Han premature infants without RDS as the control group. SP-C gene exon 5 mutation were detected using the gene sequencing method. Results SP-C gene exon 5 region c. 715G > A heterozygous mutations were detected in 17 cases among 60 patients in the RDS group. The mutation frequency was 28. 3% . SP-C gene exon 5 region c. 715G > A heterozygous mutations were detected in 8 cases among 60 patients in the control group. The mutation frequency was 13. 3% . The mutation frequency in the RDS group was statistically significantly higher than the control group (χ2 = 4. 093,P =0. 043) . In RDS group, c. 715G > A heterozygous mutation had no significant correlation with RDS grades, oxygen therapy, pulmonary surfactant dose nor treatment outcome (P > 0. 05). Conclusions A correlation may be existed between SP-C gene exon 5 area c. 715G > A heterozygous mutation and RDS in premature infants.

Objective To investigate the effects of aging on procollagen α polypeptide gene transcription and protein expression in rat vascular smooth muscle cells.Methods Vascular smooth muscle cells from thoracoabdominal aorta in neonate and 9 months old healthy Wistar rats were cultured in vitro.Results Transcription polymerase chain reaction(RT PCR) and Western blotting were used to detect type Ⅰ and Ⅲ pro-collagen α polypeptide mRNA and protein.The RT-PCR displayed that type Ⅰ procollagen α polypeptide mRNA expression had no significant difference between young group and adult group [(76.62±1.05) vs.(78.37±2.42),P＞0.05].Type Ⅲ procollagen α polypeptide mRNA expression was (105.40 ± 2.66) in young group and (123.10 ± 3.81) in adult group(P＞0.05).Type Ⅰ procollagen α polypeptide mRNA expression was (3.13 ±0.54) in young group and (4.63 ± 1.03) in adult group (P=0.05).Type Ⅲ procollagen α polypeptide mRNA expression had no significant difference between the adult and young groups[(6.86 ±0.41) vs.(7.68±0.63),P＞0.05].Type Ⅰ and type Ⅲ procollagen α polypeptide protein expressions were increased significantly in adult group as compared with the young group [(0.10 ± 0.03) vs.(0.06±0.03),(0.58±0.06) vs.(0.40±0.02),both P＜0.05].Conclusions Aging increases the procollagen α polypeptide level in vascular smooth muscle cell,which may involve in the development of vascular remodeling and atherosclerosis.

Aged ; Carcinoid Tumor ; pathology ; Humans ; Laryngeal Neoplasms ; pathology ; Male ; Neoplasm Metastasis

Aged ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Carcinoma, Merkel Cell ; metabolism ; pathology ; Carcinoma, Small Cell ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Lymphoma ; metabolism ; pathology ; Melanoma ; metabolism ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Synaptophysin ; metabolism

To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals ; Animals, Genetically Modified ; genetics ; Female ; Fibroblasts ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Goats ; genetics ; Humans ; Lactoferrin ; genetics ; Lactoglobulins ; genetics ; Milk ; chemistry ; Nuclear Transfer Techniques ; Plasmids ; Pregnancy ; Transfection

Objective To evaluate the feasibility of reconstructing horizontal periodontal bone defects by tissue engineering based on bone marrow stromal cells(BMSCs)as seed cells and enamel matrix proteins(EMPs)as growth factors. Methods Two healthy rhesus monkeys were selected, and BMSCs were isolated from iliac marrow and serial subcultivation was conducted. The cells of induced BMSCs at passage 3 were harvested and mixed with Bio-oss collagen. The models of horizontal periodontal bone defects were established surgically in each buccal side of the posterior teeth, and were divided into four groups (blank control group, material group, cells/material group and cells/material/EMPs group). The histological and Micro-CT observation were carried out 8 weeks later. Results In the blank control group, the defects were filled with fibrous connective tissue. There was newly-formed alveolar bone in the material group. In the cells/material group, periodontal regeneration could be observed, while the newly-formed cementum was irregular and less in quantity. In the cells/material/EMPs group, the amount of newly-formed alveolar bone was larger, and the newly-formed cementum was continuous and regular. Conclusion The tissue engineering technique of BMSCs as seed cells in combination with EMPs induction can significantly promote the regeneration of periodontal tissue.

Objective To investigate the reversal effect of MDR1 and MDR3 gene silencing on resistance of A2780/taxol cells to paclitaxel.Methods shRNA plasmid vector specifically targeting MDR1 and MDR3 genes was transfected into A2780/taxol cells.The early stage cell apoptosis and the effect of intracellular rhodamine 123(Rh123)accumulation were detected by flow cytometry(FCM).The late stage cell apoptosis rate was detected by terminal deoxynucleotidyl transferase(TdT)-mediated deoxyuridine triphosphate(dUTP)nick end labeling(TUNEL).The 50% inhibition concentration(IC_(50))of paclitaxel on A2780/taxol cells was determined by methyl thiazolyl tetrazolium(MTT)assay.MDR1 and MDR3 mRNA were assessed by RT-PCR,and caspase-3 protein was detected by western blot.Results After treatment with MDR1 and MDR3 shRNA plasmid vector,early apoptosis rate of A2780/taxol cells was (20.21?0.56)% and(10.87?1.24)%,respectively.MDR1 and MDR3 shRNA could increase cellular Rh123 accumulation(116.6?8.1 and 98.4?3.8,respectively).The late stage apoptosis rates detected by TUNEL displayed the same tendency as FCM results did.The IC_(50)for paclitaxel of A2780/taxol cells was decreased significantly.The mRNA levels of MDR1 and MDR3 in A2780/taxol cells were decreased by (73.3?0.8)% and(51.6?0.4)% of control,and the reduction of MDR1 and MDR3 mRNA was in a time-dependent manner.The expression of caspase-3 protein of MDR1 and MDR3 shRNA vector transfected group in A2780/taxol cells was significantly increased [(80.8?2.6)% and(72.0?4.7)%, respectively ].Conclusion MDR1 and MDR3 gene silencing could recover sensitivity of A2780/taxol cells to paclitaxel and induce cell apoptosis,thus reversing cell resistance to paclitaxel.

OBJECTIVETo investigate the toxicological mechanism of microcystin-LR (MCLR) on L-02 cells.

METHODSL-02 cells was treated with MCLR at different concentrations and the subsequent changes such as cell proliferation (MTT assay), morphology, lactate dehydrogenase (LDH) leakage, apoptosis rate and apoptosis-related gene expression were examined.

RESULTSMTT assay showed that MCLR mildly inhibited the cell growth within the initial 24 h of treatment but enhanced the cell viability after that till 60 h in a time- and dose-dependent manner. LDH leakage underwent no marked changes in response to 48-hour MCLR treatment but increased upon prolonged treatment for 60 h, indicating the presence of oxidative damage. After a 48-h treatment with MCLR at 50 microg/ml, obvious apoptosis of L-02 cells occurred as manifested by cell rounding, detachment from the substrate, cell shrinkage and membrane blebbing. The apoptosis rates were rather low (between 22% and 29%) after treatment with MCLR at different concentrations for 36 h, and increased to as much as 80% after a 60-h treatment with 50 microg/ml MCLR. The expressions of p53 and bcl-2 increased in the cells after treatment with high-concentration MCLR, suggesting that MCLR up-regulated the expression levels of the two proteins.

CONCLUSIONMCLR can induce apoptosis and up-regulate p53 and bcl-2 expressions in human normal liver cell line L-02.

Apoptosis ; drug effects ; Cell Line ; Hepatocytes ; cytology ; Humans ; Microcystins ; toxicity ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics