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.

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.

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.

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 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.

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.

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

Objective To investigate the value of contrast-enhanced ultrasound(CEUS) to diagnose thyroid nodules with different calcifications,and analyze its correlation with age and gender.Methods The image characteristics of high frequency ultrasound and pathological results of 522 patients with calcified thyroid nodules were analyzed retrospectively.CEUS enhancement patterns of different age and gender were observed and compared.Results There were no malignant nodules in patients under the age of 10.In the group of male and female,malignant nodules patients constituted the highest proportion between 40-49years old and a minimal proportion between 10-19 years old.The CEUS for weak enhancement with micro calcifications were mostly malignant nodules;the high or equal enhancement with macrocalcifications were mostly benign nodules.But a part of micro calcifications in young women patients (20 to 29 years old) with high or equal enhancement were mostly malignant nodules.Conclusions The CEUS for weak enhancement with microcalcifications are mostly malignant nodules,but a part of microcalcifications in young women patients (20 to 29 years old) with high or equal enhancement are mostly malignant nodules.

Objective To investigate the effect of intermittent hypoxia (IH) with pulmonary emphysema on the ex-pression of hypoxia inducible factor-1α(HIF-1α),Bax and Bcl-2, and the mechanism underlying the role of HIF-1αin he-patocyte apoptosis thereof. Methods Sixty rats were randomly divided into four groups: normal control group, rats were treated normally;IH group, rats were treated by 30 s nitrogen and then 90 s air, and rats were treated by from 9:00-17:00 daily;pulmonary emphysema group, rats were treated by smudging for half an hour, twice a day (8:00 and 18:00);IH with pul-monary emphysema group, rats were treated by 30 s nitrogen and then 90 s air from 9:00-17:00 daily. After exposure four-teen weeks, rats were killed. qRT-PCR assay was conducted to detect the expression of HIF-1α mRNA, Bax mRNA and Bcl-2 mRNA in live tissues. Results The expressions of HIF-1αmRNA, Bax mRNA and Bax/Bcl-2 were significantly higher in IH with pulmonary emphysema group than those in control group,IH group and pulmonary emphysema group (P<0.05). The expression level of Bcl-2 mRNA was significantly lower in IH with pulmonary emphysema group than that of con-trol group and pulmonary emphysema group (P < 0.05), but no significant difference compared with that of IH group (P >0.05). The levels of HIF-1αand Bax were positively correlated with the level of Bax/Bcl-2 (r=0.732 and 0.699),but the lev-els of HIF-1αand Bax were negatively correlated with the level of Bcl-2 (r=-0.705). Conclusion The expression levels of HIF-1αmRNA, Bax mRNA and Bcl-2 mRNA were over-regulated in hepatocytes induced by intermittent hypoxia with pul-monary emphysema. The HIF-1αexpression was correlated with Bax and Bcl-2, suggesting that HIF-1αmay promote the hepatocyte apoptosis through transcriptional co-activators, Bax and Bcl-2.

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.