Objective To investigate the alteration of protein kinase C (PKC) and endothelin system in early diabetic rats, and the effect of specific PKC inhibitor on the expression of retinal endothelin-1 (ET-1). Methods The rats model with streptozotocin(STZ)-induced diabetes were set up. The expression of retinal PKC was detected by enzyme-linked immunoabsorbent assay (ELISA). The expression of retinal ET-1, ET-3, ET-A and ET-B receptor mRNA was determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The alteration of retinal ET-1 mRNA after intravitreal injection of PKC inhibitor GF109203X in diabetic rats was also observed. Results The activities of membranous PKC were significantly increased in 2-week diabetic rats compared with that in normal rats(t=3.296, P=0 008), while activities of cytosolic PKC were unchangeable(t=0 138, P=0 894). The expression of retinal ET-1 mRNA was significantly increased(P=0 008), while no change was found in expression of ET-3, ET-A and ET-B mRNA(P=0 918,P=0 889,P=0 500). After intravitreal injection of 10 -5、10 -6、10 -7 mol/L PKC inhibitor GF109203X in diabetic rats, the expression of retinal ET-1 mRNA was decreased in a dose-dependent manner compared with the control rats. Conclusion Activation of PKC and increased expression of ET-1 could be found in the retina of early diabetic rats, and PKC inhibitor could inhibit the expression of retinal ET-1.

BACKGROUND: Bone marrow mesenchymal stem cell (MSC) is a kind of stem cell with potential of self-repair and multi-differentiation. It may differentiate into neuron, adipose cell and osteoblasts.OBJECTIVE: To observe the transforming efficacy of human bone marrow mesencymal stem cells (hMSCs) into neurons in vitro in different generations so as to provide reliable experimental data for the clinical application of MSCs.DESIGN: Single sample was designed.SETTING: Department of Neurology, Sino-Japan Friendship Hospital,Jilin UniversityPARTICIPANTS: Marrow tissue was collected from 9 cases of spinal fusion in Department of Orthopedics of First Hospital affiliated to Jilin University. Of 9 cases, all of them were in known of the experiment.METHODS: The experiment was performed in Sino-Japan Hospital affiliated to Jilin University from September 2002 to February 2003. The primary and generative culture of hMSCs was given. Experimental and the control groups were divided.-mercaptoethanol was taken as inducer. hMSCs of the 2rd, 4th, 6th and 8th generations were selected for induction in vitro for 6 h. Cytochemistry staining and immunohistochemistry staining were used to assay the expressions of neuronal and astrocytic marked proteins.MAIN OUTCOME MEASURES: ① Growth curve analysis on genera tive culture of hMSCs. ② Nissel staining. ③ Immunohistochemical staining.RESULTS: ① Common characters of generative culture: The latent phase of generative culture was 12-24 hours, exponential phase was 7-10 days and 11-13 days later, cell culture entered the platform phase. ② After induction of the 2nd, 4th and 6th generated hMSCs, deep blue granular Nissl body presented in cytoplasm. In 6 hours on the 8th induction, there was no obvious deep blue Nissl structure presented in cytoplasm.③ Except GFAP, NSE and NF-M were expressed in hMSCs of different generations after induction for 6 hours. There was no significant difference in positive rates of the 2nd, 4th and 6th generations (P ＞ 0.05), but the significant difference presented in comparison between the 8th generation and the 2nd, 4thand 6th generations (P ＜ 0.05). CONCLUSION:-mercaptoethanol can induce hMSCs differentiating into neuronal cells in vitro. The positive rates of the 2nd, 4th and 6th generationsare higher remarkably than the 8th generation.

Objective To identify and confirm a novel HLA allele.Methods A new human leukocyte antigen A allele was found during routine HLA genotyping by polymerase chain reaction-sequence specific oligonucleotide probes(PCR-SSOP) and sequencing-based typing (SBT).HLA-A locus was amplified from exon 1 through exon 8,and the nucleotide sequence of exon 2 to exon 4 for HLA-A were sequenced in both directions.Results The novel HLA-A * 31 allele is identical to A * 31 ∶ 01 ∶ 02 with an exception of one base substitution at position 245 of exon 2 where an ' A' change to ' C' resulting in codon 82 changed from GAG to GCG.Conclusion A novel HLA allele,A * 31 ∶ 22,was identified,and was named officially by the WHO Nomenclature Committee for factors of the HLA system.

Objective:To express and purify recombinant human collagen type Ⅲ and evaluate its properties.Methods:The recombinant genetic engineering strain pET30a(+)-1880/pACYCDuet-hy726/bL21(DE3) was constructed to stably co-express recombinant human type Ⅲ collagen (rhCol) and prolyl hydroxylase. rhCol was prepared and purified by E. coli high-density fermentation, salting out and column chromatography protein purification technology. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to determine the purity of rhCol. The N-terminal amino acid sequences of rhCol were determined by automatic protein polypeptide sequencing instrument. The hydroxyproline content of rhCol was determined by ultraviolet spectrophotometry. The cellular compatibility of rhCol was evaluated by MTT assay. Results:The final wet weight of high-density fermentation was about 200 g/L. The expression level was about 3 g/L. The purity of rhCol by affinity chromatography was over 95%. The results showed that the hydroxyproline content of rhCol was 11.44%, and the rhCol products have good water solubility and cell compatibility.Conclusions:RhCol can be widely applied to the field of skin care and biomedicine as an excellent biological material.

Visfatin was recently reported as an adipokine and was found to exert insulin-mimicking effects.The results showed that the expression of visfatin parallelled with obesity and insulin resistance in long term high fat chow-fed rats.The expression of visfatin mRNA was decreased and the insulin resistance improved after rennin-angiotensin system was blocked.Visfatin may play an important role in the pathogenesis of insulin resistance.

Objective To discuss the clinical efficacy of video-assisted thoracoscopic surgery(VATS)for bilateral bullous lung disease operation.Methods VATS procedures were performed on 120 patients with bilateral ballous lung disease.56 patients were operated in one-stage with traditional incision surgery.Results 120 patients were cured.The mean duration of chest drainage after surgery was 4.7 days.Postoperative complications occurred in 14 cases,including pulmonary edema,pneumothorax again respiratory failure and so on.The mean postoperative hospital stay was 14.5 days.The patients were followed up 1 to 60 months.Conclusion VATS is an effective and preferred method for bilateral bullous lung disease.compared with conventional open techniques for bilateral bullous lung disease,the advantages of VATS include shorter hospital stays,less postoperative pain and earlier return to work.

Objective To observe the effect of exposure of emphysema with intermittent hypoxia on oxidative stress injury of myocardial cells in rats. Methods Sixty male Wistar rats were divided randomly into four experimental groups(each n=15). The normal control group was bred normally. The emphysema group was exposed to cigarette smoke twice a day(once 30 minutes). The intermittent hypoxia(IH)group was exposed to intermittent hypoxia circumstance 8 hours/day,and the emphysema with IH group was exposed to cigarette smoke twice a day (once 30 minutes)and intermittent hypoxia circumstance 8 hours/day. Each group was exposed for 8 weeks. At the beginning of 9 weeks,the blood gas analysis was performed in 5 rats selected randomly from each group,and the rest rats were sacrificed and their hearts and lungs were taken. Under light microscope,the lung tissues stained with hematoxylin-eosin(HE)were examined. The lung pathology and the results of blood gas analysis showed that the emphysema with IH rat model was established successfully. The levels of malonaldehyde(MDA)and superoxide dismutase(SOD)in rat myocardium were measured by enzyme-linked immunosorbent assay(ELISA),and the subunit p22phox mRNA expressions of nicotinamide adenine dinucleotide phosphate(NADPH)-oxidase were detected by real-time reverse transcription-polymerase chain reaction(RT-PCR). Results Compared with the normal group, the MDA levels and p22phox mRNA expressions were increased obviously in emphysema group, IH group and emphysema with IH group〔MDA(μmol/g):2.93±0.54, 3.58±0.63, 4.51±0.72 vs. 1.75±0.56, p22phox mRNA:0.043±0.004,0.067±0.015,0.123±0.016 vs. 0.018±0.002,all P<0.05〕,but the activities of SOD were decreased significantly(U/mg:36.07±4.79,33.51±7.12,24.29±5.36 vs. 46.08±5.12,all P<0.05). In emphysema with IH group,the increase of MDA levels and p22phox mRNA expressions and decrease of SOD levels were more remarkable compared with those in emphysema group and IH group(all P<0.05). The expression of p22phox mRNA was positively correlated with MDA level(r=0.734,P<0.001). Conclusion The myocardial tissue oxidative stress injury in rats induced by emphysema with intermittent hypoxia exposure is more serious than that induced by exposure of either emphysema or intermittent hypoxia alone,NADPH oxidase possibly being the important medium of myocardial cell response to oxidative stress.

OBJECTIVETo investigate serological and genetic characteristics for an individual with cisAB06, an ABO blood subtype.

METHODSAntigens on red blood cells from the ABO blood group discordant individual were validated by monoclonal antibodies. The ABO antibodies in serum were validated by standard A, B, O cells. ABO genes were analyzed by polymerase chain reaction-sequence specific primer (PCR-SSP). Exons 6-7 of A/B genes were amplified with specific PCR, and the products were directly sequenced.

RESULTSBoth A and B antigens were detected on red blood cells from the proband. There was also anti-A antibody in the serum. The result of PCR-SSP has suggested a B/O02 phenotype. Direct sequencing revealed that the gene was cisAB06. Compared with B101 allele, the cisAB06 allele featured a single nucleotide change (G>C) at position 526 which resulted in an amino acid substitution (G176R).

CONCLUSIONG>C at nt526 of B allele can produce a cisAB06 allele. The serological phenotype of the specimen is therefore type AB.

ABO Blood-Group System ; genetics ; immunology ; Adult ; Alleles ; Base Sequence ; Female ; Humans ; Molecular Sequence Data ; Mutation ; Sequence Analysis, DNA

Actins ; metabolism ; Aged ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Fasciitis ; metabolism ; pathology ; Female ; Fibroma ; metabolism ; pathology ; surgery ; Humans ; Keratin-5 ; metabolism ; Mastectomy, Modified Radical ; Neoplasms, Muscle Tissue ; metabolism ; pathology

Purpose To investigate the expression of the protein of Ezrin and AQP5 in the normal, dysplasia gastric and carcinoma of esophagogastirc junction tissues and to find out their relationship with the biologic behavior of the carcinoma of esophagogastirc junction in the southern area of Hebei Province. Methods Ezrin and AQP5 proteins were detected by immunohistochemical staining in 10 ca-ses of normal gastric tissue, 40 cases dysplasia and 165 cases of the carcinoma of esophagogastirc junction tissues, respectively. Re-sults All of the expression levels of Ezrin and AQP5 protein were increasing according to the order of normal-dysplsia-carcinoma. The expression rate of Ezrin was 10%, 45% and 72. 7%, while the rate of AQP5 with 20%, 32. 5% and 65. 5% in the normal, dysplasia and carcinoma subgroup. The expression rates of each protein were significantly different among the three groups (P<0. 05). The ex-pression of them in the subgroup of poorly-differentiated with serosa invasion and lymph nodes metastasis was significantly higher than the other subgroup of well-differentiated with non serosa invasion or lymph nodes metastasis (P<0. 05). There was a positive correla-tion between the expression of Ezrin and AQP5 proteins in the carcinoma of esophagogastirc junction (P<0. 05). Conclusions Over-expression of Ezrin and AQP5 is detected in the carcinoma of esophagogastirc junction, with positive relationships, which probably helps the invasiveness and metastasis in the carcinoma of esophagogastirc junction synergistically.