Objective To observe the transfection efficiency of transferring plasmids pEGFP-N1 and pIRE-EGFP encoding enhanced green fluorescent protein(EGFP) to rabbit corneal endothelial cells(RCECs) by cationic polymer and the change of cell activity by detecting the activity of Na+-K+-ATPase.Methods RCEC was cultured by digestion and identification by neurone specific enolase(NSE) stain.Sofast TM mediated plasmid pEGFP-N1 and pIRE-EGFP to transfect RCECs and we compared the transfection efficiency at different time and in different groups by calculating the number of fluorescent cells.We observed the activity change of transfection and non-transfection RCEC by detecting the activity of Na+-K+-ATPase.Results After pEGFP-N1 and pIRE-EGFP transfected RCEC with Sofast TM by different ratio,RCEC expressed EGFP in 24-48 h.After transfection 48 h the group of SofastTM∶pEGFP-N1=3.2∶1 had the max transfection efficiency 3.6% and the max transfection efficiency of group SofastTM∶pIRE-EGFP =3.2∶1 was 3.5%,at 24 h after transfection.There were no obvious differences between the transfection and non-transfection groups of cell number and activity of Na+-K+-ATPase.Conclusion Sofast TM can mediate exogenous gene trasfering to RCEC effectively.pEGFP-N1 and pIRE-EGFP are the suitable non-virus vectors for gene transferring to RCECs.

Objective To compare the therapeutic effect of three treatment regimens on H 22 tumor-bearing mice. Methods H22 tumor-bearing mice were treated by local injection of compound chemotherapeutic agents (5-FU, mitomy-cin and cisplatin ) , oral administration of sorafeni , or the both combined , respectively .Standardized and quantitative syn-drome differentiation methods were used to assess the syndromes and tumor-inhibition rate in the mice .Results All these three therapeutic regimens were effective in suppressing the tumor growth in mice .Among them, the combined therapeutic regimen of local injection of compound chemotherapeutic drugs plus oral administration of sorafeni was the best .Conclu-sion The effect of sorafenib combined with local injection of compound chemotherapeutic agents is better than the other two regimens used separately .

Objective To study the effect of tumor size on the expression of adrenal cholesterol homeostasis mole -cules in H22 hepatoma-bearing mice.Methods Two hundred and twenty mice were injected with H 22 hepatoma cells to their right armpit.On the 11th day after injection, the mice were sorted according to the tumor size .18 mice with large tumor (large tumor group) and 18 mice with small tumor (small tumor group) were sacrificed, and the tumors were weighed.A control group consisting of 18 normal Kunming mice was also included in this study .The plasma TC, TG and HDL-C were detected using total cholesterol , triglycerides or HDL-C assay kits , respectively .The mRNA expressions of Srb1, Ldlr, Npc1, Npc2, Stard3, Hmgcr, Lipe, Acat1, Abca1, Abcg1, Srebp-1c, Lxrα, Lxrβ, Rxrα, Apoa1 and Apoe were tested by real-time quantitative RT-PCR ( qRT-PCR ) , and Gapdh and β-actin were used for normalization .SRB1 and ApoA1 proteins were analyzed by Western blot .Results The tumor weight was significantly higher in the large tumor group than that in the small tumor group (P<0.05).Compared with the control group , the plasma HDL-C was significant-ly decreased in the two hepatoma groups (P<0.05).The expression levels of Srb1, Ldlr, Apoa1 mRNA and SRB1 protein were significantly increased in the large tumor group (P<0.05 for all).The ApoA1 protein level was significantly higher in the large tumor group than that in the small tumor group (P<0.05).The expressions of Acat1, Lipe, Abca1 and Abcg1 mRNA were significantly lower in the large tumor group than those in the small tumor group (P<0.05 for all).However, the expressions of Srebp-1c, Lxrαand RxrαmRNA were not significantly changed , then, Srebp-1c, Lxrβand RxrαmRNA expressions were significantly up-regulated in the small tumor group (P<0.05).The expressions of Hmgcr and Apoe mR-NA were not significantly different in the two groups .Conclusions In hepatoma-bearing mice , due to the adaptation to tumor-induced chronic stress response , the adrenal cortical cells can effectively utilize intracellular cholesterol to synthetize cortical hormones .

Objective To evaluate the value of high frequency ultrasound in the postoperative rehabilitation treatment of rotator cuff tear.MethodsEighty-seven patients suspected as rotator cuff tear were detected by ultrasound.Fifty four patients suffered from part or entire tear were gone under the knife.Twenty-seven patients were control group with the autonomic functional training,the other twenty-seven patients were rehabilitation group with the regular rehabilitation training,Clinical functional and ultrasound scoring were performed through the two groups to evaluate the effects of rehabilitation treatment.Efficacy evaluation assessments were taken one year after operation.ResultsThe clinical symptoms were consistent with the ultrasonographic results in different time after operation.The clinical functional and ultrasound scores in rehabilitation group had no statistical difference with control group when 2 and 4 weeks after operation (P ＞ 0.05),the clinical functional and ultrasound scores in rehabilitation group were better than control group when 8,12 and 16 weeks after operation( P＜0.05 or P＜0.01 ).The efficacy evaluation assessments in rehabilitation group were much better than control group(P＜0.01).ConclusionsHigh frequency ultrasound has great significance in the postoperative rehabilitation treatment of rotator cuff tear.

It is a brief review of our research on the role of metallothioneins (MTs) in the occurrence and development of hepatocellular carcinoma (Hcc) in more than a decade,and the mechanism of prevention and treatment for Hcc in the field of TCM.It was found that MTs were required for the malignant proliferation of Hcc cells and their expressions significantly increased under different levels of stress.MTs may exert their effects on promoting proliferation via increasing ribosomal protein gene or other gene expressions.The over-expressions of MTs directly altered the balance of ions in the cell nucleus,and interacted on the genes promoting malignant proliferation.Hepatotoxicity and hepatic damage caused by DEN were reduced and relieved to different extents by the representative formula or constitute,which under the common therapeutic principle,by which MTs expressions were down-regulated as well.In conclusion,both effects above are important in the liver protection,inhibition of hyperplasia and carcinogenesis.

Objective On the issues of students' lack of representative collection of literature in problem-based learning (PBL) courses of TCM integrated experiments, to try to explore the litera-ture studying and reading route. Method Totally 41 full-time seven-year medical postgraduate of Shanghai University of TCM were randomly divided into two PBL classes, that are class A (n=20) an class B (n=21). Three PBL units were carried out in class A and class B by parallel; the oriented literatures were provided by tutors alternately, and after PBL courses, the appraisal were performed by students and tutors independently, using SPSS 19.0 for data analysis, t test to compare between groups. Results First, the students' assessment indicated that the 75% (15/20) students of Class A and 80.9%(17/21) students of Class B of class gave a good evaluation of the teaching method. Two classes had more than 80% of the students who thought the literature teachers provided was representative and instructive and over 60.0% students of both classes thought that the teaching measures could improve the classroom efficiency and after-school learning efficiency. The tutors' assessment revealed that there were no significantly differences between the quantity of these references in PPT (P>0.05) made by students in A class and B class in PBL unit 2 and unit 3; but an increasing trend did exist after the leading literatures were provided and the discussions among students were focusing and rele-vant and the teaching goal was achieved . Conclusion The route of providing a small amount of oriented literatures by tutor is positive in PBL courses of TCM integrated experiments, while the design of teaching experiment, the specific way of providing oriented literatures, the quantity and quality of literatures, and even the internal connection between them, all needs further optimization.

Objective To explore the effect of modified type B lynch uterine suture combined with calcium in the prevention and treatment for uterine atony postpartum hemorrhage.Methods 63 pregnant women with postpartum hemorrhage risk factors which would be underwent cesarean delivery were randomly divided into the study group (32 cases) and the control group (31 cases).The study group was given intravenous calcium gluconate (10ml) 10 min before the operation,and modified type B lynch uterine suture in the operation.The control group was only given the same dose of oxytocin of the study group.Results The amount of bleeding intraoperation and 24h after the operation was (182.6 ± 47.7) ml and (214.1 ± 63.8) ml,respectively,the time of intraoperative uterine contractions was (3.8 ± 1.3)min,the degree of hemoglobin decreased 24h after the operation was (9.2 ±2.0) min in the study group,which were less than those of the control group(t =7.26,2.97,2.39,4.01,all P ＜ 0.05).There was no statistically significant difference in puerperal morbidity and neonatal asphyxia between the two groups (all P ＞ 0.05).Conclusion The modified type B lynch uterine suture combined with calcium could effectively prevent and treat uterine atony postpartum hemorrhage,and could not increase the rate of neonatal asphyxia and puerperal morbidity.

Objective To study the expression features of hydrolase genes related to the secretion of thyroid hormone of H22 hepatoma mice with different symptoms in early stage. Methods Firstly, The quantitative diagnosis and syndrome differentiation methods were used in H22 tumor-bearing mice in early stage, the expression profile of Tg and related hydrolase genes in poisonous pathogenic factors syndrome group (PPFS) and qi-deficiency syndromes (QDS) were got, and the major differential expression were selected. Secondly, the experiment was repeated and ELISA were used to detect T3 and T4 in serum, RT-PCR were applied to detect gene transcription level of genes including Tg, Ctsb, Ctsd, Ctsl, Napsa and Tpp1. Results ① Based on gene chip, the expression of Tg, Ctsb, Ctsd, Ctsl, Napsa and Tpp1were decreased in the first batch of experiment, the exactly ratio was Tg(0.77 in PPFS;0.84 in QDS), Ctsb(0.83 in PPFS, 0.91 in QDS), Ctsd(0.79 in PPFS;no notable change in QDS), Ctsl(no notable change in PPFS; 0.65 in QDS), Napsa(0.78 in PPFS; no notable change in QDS), and Tpp1 (0.75 in PPFS; no notable change in QDS), respectively. ② T3 and T4 downregulated in PPFS (the T3 value was 1.519±0.162ng/ml, T4 value was 2.194±0.305mg/dl) and in QDS (the T4 value is 4.366±0.727μg/dl) in early stage (P<0.01), especially in PPFS, which was in accordance with the change of Tg in both batches. ③the same trend happened in the validation of Tg(0.22 in PPFS;0.38 in QDS), Ctsb(0.31 in PPFS;0.55 in QDS), Ctsd(0.36 in PPFS;0.78 in QDS) and Napsa(0.24 in PPFS;0.59 in QDS) ,while ctsl(1.24 in PPFS;2.11 in QDS) and Tpp1 (2.85 in PPFS;0.85 in QDS)werethe opposite;even this, the total trend of the expression in QDS was still higher than that in PPFS. Conclusion All the results showed that the thyroid function of H22 hepatoma mice was inhibited in early stage especially in PPFS.

OBJECTIVE: To reveal the characteristics of gene expression in adrenal gland of H22 tumor mice with typical syndromes and in different liver cancer stages. METHODS: By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array, we observed adrenal gland gene expression in H22 tumor mice with pathogenic factor-toxin predominance syndrome and qi deficiency syndrome in the earlier stage, yang-qi deficiency syndrome in the intermediate stage, and qi-yin-yang deficiency syndrome in the advanced stage. Genes highly expressed and remarkably different were analyzed in this study. RESULTS: A total of seventy-three up-regulated coincident genes and twenty-six down-regulated coincident genes in different stages were investigated in the study. Up-regulated coincident genes included Hp, C3, Anxa1, Procr, C2, Il4ra, Cd14, Ptprc, Cd52, C4b, Eno3, Xdh, Gpx3, and so on. Down-regulated coincident genes included nervous system function-related genes such as Plp1, Mbp, Aldh1a1, Cck, Atn1, genes associated with electrolyte metabolism such as Aldh1a1 and Slc22a17, genes related to signal transduction such as Cxcr4, Spag5 and Stmn3, etc, and genes related to transcriptional control and protein biosynthesis such as Hspa1a, Dnajb1, Thra, Hhex and so on. CONCLUSION: With the development of the tumorigenesis, the symptoms and signs and differentially expressed genes in adrenal gland of H22 tumor mice can be measured. Up-regulated and down-regulated coincident genes may be the features of H22 tumor mice different from those of normal mice.

Objective:To understand the genotyping and molecular epidemiological characteristics of Norovirus (NoV) GⅡ in adult acute gastroenteritis outpatients in Shanghai from 2015 to 2016.Methods:A total of 912 stool specimens from adult patients with acute diarrhea from March 2015 to December 2016 (431 in 2015 and 481 in 2016) were collected. The Norovirus GⅡ type was detected by one-step quantitative reverse transcription PCR assay and the RdRp region of open reading frame (ORF) 1 and 5′end of ORF2 were amplified. The region was sequenced and classified.Results:From March 2015 to December 2016, NoV GⅡ were detected in 17.76% (162/912) of the samples, 15.08% (65 /431) in 2015 and 20.17% (97/481) in 2016. Based on sequence analysis of the RdRp and capsid sequences, 145 identified NoV strains were divided into 10 genotypes: GⅡ.Pe_GⅡ.4 Sydney 2012 (60), GⅡ.P17_GⅡ.17 (45), GⅡ.P16_GⅡ.13 (10), GⅡ.P12_GⅡ.3 (10), GⅡ.P7_GⅡ.6 (9), GⅡ.P21_GⅡ.21 (5), GⅡ.P16_GⅡ.4 Sydney 2012 (2), GⅡ.Pe_GⅡ.17 (2), GⅡ.P2_GⅡ.2 (1), and GⅡ.P7_GⅡ.14 (1).Conclusions:The main epidemic NoV GⅡ genotype in Shanghai was GⅡ.P17_GⅡ.17 in the spring of 2015. GⅡ.Pe_GⅡ.4 Sydney 2012 and GⅡ.P17_GⅡ.17 were identified as predominant genotypes during the winter of 2015 and spring of 2016. The most common genotype was GⅡ.Pe_GⅡ.4 Sydney 2012 in autumn of 2016. Continuous NoV outbreak surveillance is important for identifying changing trends in genotype distribution and emerging new strains.