Abstract In this paper,the sequences of second exon of DQA1 and DQB1 were amplified byPCR primers,and hybridized with 17 or 23 sequence specific oligonucleotide probes labeled byisotope respectively.The distribution of 10 DQA1 alleles and 17 DQB1 alleles was observed inHunan Hans population.The results showed that the most of DQAI and DQB1 alleles were ex-pressed in Hans,however,their frequencies are different from Caucasoids.

The establishment of professional master degree in pathology is a new branch of the professional degree graduate education.It is an important measure to bring the clinical pathology into the teaching system of clinical medicine specialty in China.In response to a wave of education reform in the new century, our department has positively converted the traditional postgraduate training and teaching mode, innovated the teaching idea and content, and continued to improve the teaching method and means.On the basis of fully respecting students' subject consciousness, we have combined traditional and modern teaching means, made full use of modern education information technology, actively built and promoted pathology specialized degree graduate student research, teaching and clinical trinity management: the new training mode.Through the practice of bilingual, network, clinical problems and cases of teaching and combined with the leading edge forum of famous pathologists' research and the professional standardized training of clinical pathology, we have extensively carried out the education training, which is based on the theory, practice, research, and sub specialist guidance.

Objective　To explore the effect of nordihydroguaiaretic acid (NDGA) on the expressions of vascular endothelial growth factor (VEGF) and its receptor, kinase-inserted domain containing receptor(KDR) and the possible mechanism. Methods　The expression of VEGF in human malignant glioma cell line SHG-44 and that of KDR in human umbilical vein endothelial cell (HUVEC) line ECV-304 were observed 1～3 d after NDGA treatment with immunohistochemistry, in situ hybridization and image analysis. Results　The expression of VEGF was declined at protein or mRNA levels in SHG-44 cells after treated with 100 μmol/L NDGA for 1 to 3 d. The expression of KDR in endothelial cells with 100 μmol/L NDGA treatment for 1 to 3 d was decreased too, in a more obvious way compared with the decline of VEGF expression in SHG-44 cells. Conclusion　The results suggest that NDGA inhibits the expression of VEGF in glioma cells as well as that of VEGF receptor KDR in endothelial cells, which may be the important molecular mechanism of anti-angiogenesis of NDGA.

Objective　To investigate the changes and their significance of bcl-2 and c-myc in nordihydroguaiaretic acid (NDGA)-induced apoptosis of human malignant glioma cell line SHG-44. Methods　The apoptosis of SHG-44 cells was observed with light and electron microscopy and TUNEL method. The expression of bcl-2 and c-myc gene was measured with immunohistochemistry, in situ hybridization and image analysis. Results ① The SHG-44 cell apoptosis was induced by NDGA at a concentration lower than 200 μmol/L in a time-dependent manner. ② The expressions of bcl-2 and c-myc gene in SHG-44 cells were decreased after the treatment of 100 μmol/L NDGA with the elapse of time, indicating a close association with cell apoptosis. ③ The expressions of bcl-2 and c-myc mRNA in SHG-44 cells were decreased after the treatment with 100μmol/L NDGA, which was apparently consistent with the immunohistochemical results. Conclusion　The NDGA-induced apoptosis in human malignant glioma cells might be related with the down-regulated expressions of bcl-2 and c-myc gene. The exact mechanism needs further research.

Objective　To investigate roles of cyclin-dependent kinase 4 (CDK4) in nordihydroguaiaretic acid (NDGA)-induced inhibitory effect on proliferation of human malignant glioma cells. Methods　The techniques of cell culture, cell counts, flow cytometry, immunoprecipitation, immunohistochemistry and image analysis were employed in this study. Results　①A concentration-dependent inhibition of proliferation was demonstrated in the SHG-44 cells incubated for 24 hours in the presence of NDGA, and cell proliferation was blocked in the G1→S phase. ②The activity of CDK4 was decreased apparently in the SHG-44 cells treated for 24 hours with 10 to 200 μmol/L NDGA in a concentration-dependent way. ③The expression of CDK4 gene was downregulated in the cells after NDGA treatment. Conclusion　CDK4 plays an important role in NDGA-induced inhibition of glioma cell proliferation.

75dB SPL or no response. RESULTS The individual differences of CM absolute amplitudes among the normal hearing ones at different frequencies are distinct, but bilateral CM amplitudes in same person are almost uniform and stable. Enlargedand prolongated CM was found in ears with loudness recruitment .Of 104 cases with unilateral hearing loss, CM were enlarged and prolongated at corresponding frequencies with loudness recruitment in 95 cases(91.3 %).The prognosis of cases with CM type Ⅰ was better than those with types Ⅱ and Ⅲ. The effective rates of treatment were 78.19 % in type Ⅰ , 2.5 % in type Ⅱ, and none in type Ⅲ. There were significant differences among them. CM was slightly enlarged during sleep, and the detected threshold of CM is less than that of the awake condition. CONCLUSION CM offers the reliable information for the mechanism of loudness recruitment and is also useful for understanding the relationship among loudness recruitment, cochlear microphonics and prognosis. CM may be taken as a valuable parameter for evaluating prognosis.

Objective To probe the diagnostic efficacy of fetal eehocardiography for characterizing complex congenital heart diseases.Methods Fetal echocardiography was performed on 49 cases of fetal complex congenital heart disease, the ultrasonic diagnosis was compared retrospectively with pathological results after autopsy.Results Antenatal sonographic diagnosis was in agreement with the pathological results in 42 cases (85.71 %), 7 cases were disagreed with pathological diagnosis (antenatal sonographic diagnosis was discrepancy in 3 cases, 4 cases were partially mis-classified).Twenty-four cases were combined with extra-cardiac malformations.Nine cases had chromosomal abnormality.Conclusions Fetal echocardiography is highly accurate for antenatal diagnosis of complex congenital heart disease, but it is hard to detect some type of cardiac anomalies.

Objective:To study the influence of different culture conditions on charcic and inhibition activity of nature killer cells ( NK) ,whether to join the modified K562 cells with IL-6 cytokine.Methods:According to the 5′end of the human IL-6 cDNA sequence,PCR primers designed to amplificate,express and transfect K562 cells cDNA library as a template for DNA.Genetic modified K562 cells as stimulating cells were prepared by expressing IL-6.To extract peripheral blood mononuclear cells( PBMC) from human peripheral blood.PBMC were explanted by genetic modified K562 stimulated.The expansion was initiated by CO-culture of PBMC and irradiate genetic modified K562 cell.The number of NK cell increased by directed induced generation of genetic modified K562 cell.Immunophenotypic analysis of NK cell surface markers was performed by flow cytometry (FCM).51Cr release assay was employed to measure the specific lysis skilling of NK cell target K562 cells.Results:We have constracted genetic modified K562 cells by genetic engineering.As stimulated cell added into the PBMC,an average of 760 ±18 fold expansion of CD56+CD16+CD3-cells was observed after 3 weeks of co-culture system.The NK cells population could proliferated more 91%±2% after expansion comparing with 6%± 0.4%in PBMC before expansion by FCM.The cytotoxical activity of NK cells which was induced by genetic modified K562 cell was the strongest than induced by IL-6 cytokine alone.The expanded NK cells lysed 92%±2% of K562 targets in a 5∶1 effector to target ratio.In this case,the NK cells induced by genetic modified K562 cells against tumor cells was more lethal.Conclusion:PBMC based in vitro expansion of natural killer cells was set up by genetic modified K562 cells.The cytotoxicity of NK cells was the strongest induced by genetic modified K562 cell treated.These results had important guiding significance for the the NK large number of amplification and used in clinical.

Objective:To explore the operative effect and treatment strategies for the low distal humerus fracture.Methods:A retrospective analysis was conducted on the clinical data of 16 patients with the low distal humerus fracture between January 2016 and January 2018 at Department of Orthopaedics, Shanghai Tongji Hospital Affiliated to Tongji University.All fractures were coronal fractures of humeral head, partly combined fractures of humeral trochlea or posterior part. Fractures were classified according to Dubberley classification as 9 cases in type Ⅰ, 3 cases in typeⅡ and 4 cases in type Ⅲ.Selection of the kind of operative approach and internal fixation was made according to the fracture type and shape. For simple coronal fractures of humeral head or combining humeral trochlea fractures, which were relatively stable, single or multiple countersunk screws fixation by the lateral approach were chosen.For humeral head coronal fractures, which combining obviously displaced comminuted humeral trochlea factures, posterolateral locking plates and countersunk screws internal fixation by the olecranon osteotomy approach were chosen. The incision and elbow soft tissues were observed within 2 weeks after operation.The radiographic evaluation of fracture reduction, bone healing, internal fixation, arthritis and elbow range of motion were made at 3, 6, 12 months after operation. The Mayo elbow functional scores were documented for analysis of elbow joint function, and compared between different surgery groups by Kruskal-Wallis H test. Results:The follow-up time was (22.1±9.2)months(range: 15 to 39 months). The incisions healed well in 2 weeks after operation without soft tissue infection, necrosis or vascular complications. There was no fracture reduction loss or internal fixation loosening according to radiographic evaluation 3 months after operation. One case of ectopic ossification was observed 6 months after operation and inhibited by the treatment of non steroidal anti-inflammatory drugs.One case of ulnar neuritis occurred after operation and released after removing the long screw and loosing the ulnar.Osteoarthritis images were observed at the end of follow-up.Arc of motion was (120.4±11.2) ° in flexion and (5.5±1.9) ° in extension. The Mayo score was 88.7±9.1, including 11 excellent, 4 good, and 1 fair.The Mayo score was 90.1±3.7 in Dubberley classification type Ⅰcases, 89.7±4.6 in type Ⅱ cases and 84.5±5.8 in type Ⅲ cases. There were no significant differences in Mayo scores between 3 types cases according to Kruskal-Wallis H test. Conclusion:Choosing the appropriate surgical approach and composite internal fixation according to the fracture types and shapes of low distal humerus fracture, anatomic reduction of the articular surface and early functional exercise are the keys to obtain ideal curative effect.

Objective:To explore the operative effect and treatment strategies for the low distal humerus fracture.Methods:A retrospective analysis was conducted on the clinical data of 16 patients with the low distal humerus fracture between January 2016 and January 2018 at Department of Orthopaedics, Shanghai Tongji Hospital Affiliated to Tongji University.All fractures were coronal fractures of humeral head, partly combined fractures of humeral trochlea or posterior part. Fractures were classified according to Dubberley classification as 9 cases in type Ⅰ, 3 cases in typeⅡ and 4 cases in type Ⅲ.Selection of the kind of operative approach and internal fixation was made according to the fracture type and shape. For simple coronal fractures of humeral head or combining humeral trochlea fractures, which were relatively stable, single or multiple countersunk screws fixation by the lateral approach were chosen.For humeral head coronal fractures, which combining obviously displaced comminuted humeral trochlea factures, posterolateral locking plates and countersunk screws internal fixation by the olecranon osteotomy approach were chosen. The incision and elbow soft tissues were observed within 2 weeks after operation.The radiographic evaluation of fracture reduction, bone healing, internal fixation, arthritis and elbow range of motion were made at 3, 6, 12 months after operation. The Mayo elbow functional scores were documented for analysis of elbow joint function, and compared between different surgery groups by Kruskal-Wallis H test. Results:The follow-up time was (22.1±9.2)months(range: 15 to 39 months). The incisions healed well in 2 weeks after operation without soft tissue infection, necrosis or vascular complications. There was no fracture reduction loss or internal fixation loosening according to radiographic evaluation 3 months after operation. One case of ectopic ossification was observed 6 months after operation and inhibited by the treatment of non steroidal anti-inflammatory drugs.One case of ulnar neuritis occurred after operation and released after removing the long screw and loosing the ulnar.Osteoarthritis images were observed at the end of follow-up.Arc of motion was (120.4±11.2) ° in flexion and (5.5±1.9) ° in extension. The Mayo score was 88.7±9.1, including 11 excellent, 4 good, and 1 fair.The Mayo score was 90.1±3.7 in Dubberley classification type Ⅰcases, 89.7±4.6 in type Ⅱ cases and 84.5±5.8 in type Ⅲ cases. There were no significant differences in Mayo scores between 3 types cases according to Kruskal-Wallis H test. Conclusion:Choosing the appropriate surgical approach and composite internal fixation according to the fracture types and shapes of low distal humerus fracture, anatomic reduction of the articular surface and early functional exercise are the keys to obtain ideal curative effect.