Objective: To construct the packaging plasmid pSNAV-hEndostatin-CMV-EGFP of adeno-associated virus vector (AAV) encoding human Endostatin (hEndostatin) cDNA. Methods: The hEndostatin eDNA obtained from plasmid pCD-sEndostatin was subcloned into the packaging plasmid pSNAV of AAV by molecular clone ways. The recombinant plasmid pSNAV-hEndostatin-CMV-EGFP was identified by PCR analysis, restriction enzymes analysis and sequencing analysis.Results: The recombinant pSNAV-hEndostatin-CMV-EGFP was correctly constructed and confirmed by PCR analysis, restriction enzymes analysis and sequencing analysis. Conclusion: The constructed AAV-hEndostatin packaging plasmid pSNAV-hEndostatin-CMV-EGFP could be the packaging plasmid of rAAV-hEndostatin-EGFP.

AIM: To evaluate the changes in central corneal thickness (CCF) caused by mydriasis with Mydrin-P.METHODS: A total of 106 eyes of 53 patients with refractive errors were studied. Each eye had instillation of Mydrin-P to obtain mydriasis. CCT was examined before and after mydriasis using ultrasonic pachymeter.RESULTS: CCT increased significantly after mydriasis with Mydrin-P.CONCLUSION: Mydrin-P can affect the measurement of CCT.

Objective:To investigate the effect of human interferon-α-2b(IFN-α-2b) in the treatment of endometriosis by detecting the levels of CA125 and endometrial antibody(EMAb). Methods: Forty-five cases with endometriosis were divided into three groups. Fifteen cases in test group (operation+IFN-α-2b), 20 cases in control group A (operation+Diphereline), 10 cases in control group B(only operation). The blood serum level of CA125 was detected by microparticle enzyme immunochemiluminescent at pre-operation, the first month, second month and third month of the post-operation; and the EMAb level was measured by enzyme linked immunosorbent assay (ELISA). The follow-up of patients was carried out after operation to evaluate the clinical appearances and record the adverse effects of the human IFN-α-2b. Results: Compared with pre-operation, the levels of CA125 and EMAb were obviously degraded in the first to third months after surgery in test group and control group A(P < 0.05). There were no significant difference in the levels of CA125 and EMAb were at pre-operation,the first month,second month and third month of the post-operation between the test group and control group A(P > 0.05). There were no significant difference in the levels of CA125 and EMAb at pre-operation between the test group and control group B(P > 0.05). However, the level of EMAb was lower the first month after operation in the test group than that of control group B. In the follow-up, the adverse effects of the human interferon-α-2b were observed including low-grade fever, articular muscle soreness and gastrointestinal tract complaints. These appearances completely disappeared after drug withdrawal. It had less impact on the menstruation. Conclusion: The levels of CA125 and EMAb were similar in treatment of endometriosis with human IFN-α-2b and with Diphereline. Both of the treatments are better than the pure surgery. It has less impact on the menstruation and can avoid menostasia.

Objective To investigate the clinical significance of I-FISH for detection of genomic abnormalities in MM. Methods Twenty newly diagnosed MM patients(seven cases at stage Ⅰ , five cases at stage Ⅱ and eight cases at stage Ⅲ according to Bataille staging) were analyzed by combining the technique of CC (R-binding stain) and I-FISH [ including GLP13q14 (RBI gene), GLP17p13. 1 (P53 gene),GLP13q14. 3(D13S319) ,GLP1q21 ,GLP14q32(IgH gene) DNA sequence probes]. These two methods were compared for the detection rates of chromosomal and genomic abnormalities in MM and the association between genomic abnormalities and Bataille stages was also analyzed. Results CC examination showed only 1 case [5% (1/20) ] was found complex chromosomal abnormalities--46,XX,-2,del(3) (p21) ,add(6)(q26) ,der(10)(q26),der(14)(q32), + mar, inc[6]. While I-FISH assay showed that 12 cases [60%(12/20) ] were found genomic abnormalities. The frequencies of RB1, D13S319 and P53 were all 30%(6/20), and the frequencies of IgH gene and 1q21 were both 20% (4/20). The detection rate of the I-FISH was much higher than CC (χ2 = 9. 09, P = 0. 001) according to paired χ2 test. Of 20 patients,6 cases had RB1 gene abnormality, 1 case at stage Ⅰ , 2 cases at stage Ⅱ and 4 cases at stage Ⅲ. Of 20 patients, 6 cases had D13S319 gene abnormality, 2 cases at stage Ⅰ , 1 case at stage Ⅱ and 3 cases at stage Ⅲ. Of 20 patients, 6 cases in 20 had P53 gene abnormality, 2 cases at stage Ⅰ and 4 cases at stage Ⅲ. Of 20 patients, 4 cases had 1q21 gene abnormality, 2 cases at stage Ⅰ and 2 cases at stage Ⅲ. Of 20 patients, 4 cases had IGH gene abnormality, 1 case at stage Ⅰ and 3 cases at stage Ⅲ. Conclusion Ⅰ-FISH has higher detection rate for the genomic abnormalities in MM and can be used in detection of MM patients in different Bataille stages.

Objective To investigate expression of CD56, CD95(Fas), Ki-67, p53, bcl-2, HMB45 and S-100 in mucosa amelanotic malignant melanoma in order to improve the pathological diagnosis level reduce wrong diagnosis and avoid missing dignosis, and afford objective factors for prognosis and therapy. Methods The techniques of tissues chips and immunohistochemical lablling were used for analyzing 48 cases of mucosa amelanotic malignant melanoma. Results The positive rates of HMB45 and S-100 were 100 % (48/48) and 85.4 % (41/48) respectively. The positive rate of CD56 was 91.6 % (44/48), there was not statistical difference between original cases and metastatic cases. The positive rate of CD95 was 85.4 %(41/48). In which it is 100 % (11) in 11 cases of having lymphanoid metastasis. The positive rates of Ki-67 and p53 were 79.2 % (38/48) and 58.3 % (28/48) respectively. The positive distribution of Ki-67 was almost same as CD95. The positive rate of bcl-2 was 39.6 % (19/48), the positive expression was significantly different between p53 and bcl-2(P

Objective To investigate the expression and secretion of mice DNaseI gene plasmid transfected into bone marrow (BM-MSCs) mesenchymal stem cells. Methods The plasmids of mouse DNaseI gene had been transfected into the BM-MSCs of mice by liposomes. The expression of DNaseI gene in the BM-MSCs was detected by western blotting and the DNaseI activity was measured by DNA-methyl green substrate colorimetry. Results About 30% BM -MSCs were transfected with mice plasmid DNaseI gene, DNaseI was expressed in the transfected BM-MSCs and active DNaseI could be detected in the supernatant of cell culture. Conclusion The mice DNaseI gene plasmid can be transfected into mice BM -MSCs by liposomes and DNaseI gene can be expressed by the transfected BM-MSCs and active DNaseI can be secreted. This may provide potential target for the treatment of SLE.

Objective To study the therapeutic effect of recombinant adeno-associated virus carrying human endostatin gene therapy on endometriosis in mice model. Methods Recombinant adeno-associated virus vector carrying human endostatin gene and enhanced green fluorescent proteins gene (rAAV2-endostatin-EGFP) was constructed. Endometrium was from 12 patients with leiomyoma undergoing hysterectomy in Second Hospital, Tianjin Medical University between November and December 2008. Endometriosis models of nude mice were established by transplanting human endometrial fragments intooperitoneal surface. After 1 week, those 60 mice were divided into 3 groups: treatment group including 20 mice injected with rAAV2-endostatin-EGFP to ectopic lesion, control group including 20 mice injected with rAAV2-EGFP to ectopic lesion and blank control group including 20 mice injected with phosphate buffered saline (PBS) to the ectopic lesion. At 1, 2 and 3 weeks after treatment, those mice underwent laparotemy to observe the location and size of ectopic lesion in abdominal cavity. The expression of endostain protein, number of gland, microvessel density (MVD) and vascular endothelial growth factor (VEGF) were measured in ectopic lesions. The serum level of estradiol and progesterone were detected in nude mice among every groups. Results (1) All endometriosis of nude mice models were established successfully through peritoneum transplanting. After 1 week's treatment, flat lesion nodes, decreased gland number and narrow and atrophy glandular cavity were observed by light microscope. (2) The endostatin gene was transferred into nude mice successfully and expressed effectively. It was observed that endostatin protein expression was shown with enhanced green fluorescent proteins in ectopic lesion. (3) Glands number of ectopic lesion in rAAV2-endostatin-EGFP group(7.8±1.9,7.0±1.5 and 5.5±1.7) were significantly less than 10.1± 1.7, 10.2±2.0 and 9.8±2.4 in rAAV2-EGFP control group and 10.2±2.2,10.0±2.0 and 9.7±2.2 in PBS control group at 1,2 and 3 weeks after treatment(all P<0.05). Glands number of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P<0.05). (4) MVD of ectopic lesion in rAAV2-endostatin-EGFP group (12.2±1.5,11.4±2.1 and 9.0±1.4) was significantly less than those at rAAV2-EGFP control group (16.5±1.7,16.5±1.9 and 16.9±1.9) and PBS control group (16.2±1.6,16.0±1.6 and 16.3±1.7) at 1,2 and 3 weeks after treatment (all P<0.05) . MVD of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment(P<0.05). (5) The rate and density of VEGF expression at ectopic lesion in rAAV2-endostatin-EGFP group (35%, 30%, 25% and 1.60±0. 43,1.33± 0. 30,1.03±0.36) were significantly less than those at rAAV2-EGFP control group (80% ,75% ,85% and 2.43±0.53,2.43±0.29,2.66±0.45) and PBS control group (85% ,90% ,90% and 2.36±0.53,2.64± 0.57,2.53±0.52) at one 1, 2 and 3 ,weeks after treatment (all P<0.05). The expression of VEGF at ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P<0.05). (6) The level of estradial and progesterone in serum of nude mice of rAAV2-endostatin-EGFP group [ E_(2)> : (48±7 ) pmol/L, P: (61±8 ) nmol/L ] did not reach statistical difference when compared with those at rAAV2-EGFP control group [ E_(2): (50±9) pmol/L, P: (60±10) nmol/L] and PBS control group [E_(2):(48±7)pmol/L,P: (58±10)nmol/L,P>0.05]. Conclusions The recombinant adeno-asseciated virus carrying human endostatin gene therapy could inhibit angiogenesis at endometriotic lesions and not influence steroid level. The antiangiogenic gene therapy might become a novel option for endometriosis.

Objective To analyze the related rules and prognosis analysis for lymphatic metastasis in patients for Siewert Ⅱ and Ⅲ adenocarcinoma of the gastroesophageal junction.Methods This study retrospectively reviewed 65 patients who underwent radical surgery for Siewert Ⅱ and Ⅲ adenocarcinoma of the gastroesophageal junction in Beijing Friendship Hospital between July 2013 and March 2017.The primary indicators were sex,age,tumor site,tumor size,macroscopic type,histological type,depth of invasion,operation,tumor emboli and carcinoma nodes.The clinical characteristics and pathological features were analyzed by using Logistic regression and Kaplan-Meier survival analysis.Follow-tup using outpatient examination and telephone interview was performed to detect patients' diseases and postoperative survival up to April 2017.Results Lymph node metastasis was observed in 41 of 65 patients (63.1％).Lymph node metastasis frequency was found in No.1,No.2,No.3,No.7,No.11 and No.110 lymph nodes.The incidences of those lymph node metastasis were 45.3％,32.5％,28.8％,22.5％,19.4％ and 8.2％.The metastasis rate of the tumors with diameter over 2 cm was higher than the tumors which were less than 2 cm (71.9％ vs 0;P =0.000);The observed rate of lymph node metastasis was 0,12.5％,69％,95.2％ in early cancer (T1) and advanced cancer (T2-T4) (P=0.000) respectively.The metastasis rate of highly differentiated Siewert Ⅱ and Ⅲ adenocarcinoma of the gastroesophageal junction was 45.2％ lower than poorly differentiated tumors 79.4％ (P =0.009).Thirty-three cases were found with lymphatic embolus,28 of them (84.8％) were found to be combined with lymph node metastasis among them (P =0.001).All the differences were statistically significant (P ＜ 0.05).The results of multivariable analyzed by binary logistic regression analysis revealed that neoplasms histologic type and lymphatic embolus were independent risk factors for lymph node metastasis of Siewert Ⅱ and Ⅲ adenocarcinoma of the gastroesophageal junction.Sixty-five patients were followed up for 1-45 months,with a median time of 18.81 months.Although there was no statistically significant difference,3 years-overall survival rate showed tumor without lymph node metastasis has better prognosis than with lymph node metastasis and early stage tumor also has better prognosis than advanced tumor.Conclusions Lymph node metastasis of Siewert Ⅱ and Ⅲ adenocarcinoma of the gastroesophageal junction is related with histological type of tumor and lymphatic embolus.For Siewert Ⅱ and Ⅲ adenocarcinoma of the gastroesophageal junction,the incidence of lymphatic metastasis were higher appear in abdominal cavity,which included No.1,No.2,No.3,No.7 and No.11;Meanwhile,the metastasis of mediastinal lymph node is given priority to No.110 lymph node.Therefore,radical total gastrectomy combined with standard D2 lymphadenectomy and No.110 lymphadenectomy are performed in order to improving the prognosis.For patients with lymph node metastasis and tumor in advanced stage,long-term prognosis remains to be further study.

To retrospectively analyze the safety and efficacy of low dose subcutaneous decitabine regimen in patients with acute myeloid leukemia (AML) and intermediate-or higer-risk myelodysplastic syndrome (MDS).Of 6 AML cases,2 achieved complete remission (CR),2 with partial remission(PR),1 with stable disease(SD),1 with progressive disease(PD).As to the 8 MDS patients,one achieved CR and 6 with hematologic improvement (HI),1 case SD.Low dose subcutaneous decitabine regimen could be an alternative choice of older AML or MDS patients.

Objective To improve the cognition of sever liver injury of treating Philadelphia positive acute lymphoblastic leukemia (Ph+ ALL) with salvage chemotherapy of dasatinib combined with high-dose methotrexate (HD-MTX) and L-asparaginase (L-Asp).Methods Severe drug-induced liver injury caused by dasatinib with HD-MTX and L-Asp in one patient with Ph+ ALL was reported.Results Severe drug-induced liver injury happened on the seventh day after treatment,TBIL 221.7 μmol/L,DBIL 156.1 μmol/L,IBIL 65.6 μmol/L,ALT 111 U/L,AST 131 U/L,ALP 354 U/L,GGT 256 U/L,TBA 199.2 μmol/L.Through proper treatment,the patient recovered quite good,and the patient achieved complete remission after this chemotherapy.Conclusion Salvage chemotherapy which contains dasatinib,MTX and L-Asp can be effectively used in Ph+ ALL,but they are all of the hepatotoxicity,so drug-induced Liver injury may happen while they are used together.