A hammerhead RZ DNA was designed and synthesized, which can specifically cleave the bcl-2 mRNA. After demonstration of right sequences by sequencing and cleavage activity of RZ by in vitro cleaving experiment, The RZ DNA was recombinated into the pDOR - neo vector to form the recombinant pDOR - RZ. Using lipofectin - mediated DNA transfectionpDOR-RZ was successfully introduced into HL - 60 cells. The RZ expression was observed by Southern, RNA dot blot hybridization and flow cytometry (FCM) . The results demonstrated that (a) the RZ was expressed in 72 hours after transfection; (b) the synthesis of Bel - 2 protein was inhibited by the expression of RZ; (c) apoptotic peak appeared in FCM.

Objective To study the role of down regulation of bcl 2 expression of human cholangiocarcinoma cells with bcl 2 mRNA cleaving ribozyme (RZ). Methods Synthetic gene of bcl 2 mRNA cleaving RZ was recombined with the expression vector of eukaryotic cells. The recombined vector was used to transfect human cholangiocarcinoma cells. The expression of bcl 2 in human cholangiocarcinoma cells was observed by immunocytochemistry and flow cytometry. Results The expression of bcl 2 in human cholangiocarcinoma cells transfected with RZ gene was lower than that of control group. Spontaneous apoptosis was observed in a few of the cells. Conclusion bcl 2 mRNA cleaving RZ can down regulate the expression of bcl 2 of cholangiocarcinoma cells notably and can also induce spontaneous apoptosis.

Objective To investigate the relationship between the serum ferritin levels and disease activity of patients with systemic lupus erythematosus (SLE).Methods One hundred and forty-six patients with SLE and 65 healthy volunteers were involved.Serum ferritin,C-reactive protein (CRP),erythrocyte sedimentation rate (ESR) and anti-double-stranded DNA antibodies (dsDNA) were measured in two groups.The activity of SLE was evaluated by systemic lupus erythematosus disease activity index (SLEDAI) score.The data were recorded and analyzed.SPSS 19.0.statistical software was used in statistical analysis.Analysis of variance was employed in comparison between groups using,t test was used in further pairwise comparisons,Pearson correlation test was adopted to evaluate the correlation between two groups.Results The level of serum ferritin in patients with SLE group was significantly higher than that of control group (505.4 ±408.9) ng/ml and (72.4 ±42.8) ng/ml,respectively,t =6.67,P ＜0.01.57.5％ (84/146) patients with SLE had elevated serum ferritin.Patients with high SLEDAI scores had significantly higher ferritin concentrations than other patients (807.6 ± 412.3) ng/ml and (96.0 ± 44.7) ng/ml,t =6.56,P ＜0.01.The levels of serum ferritin in SLE patients were positively correlated with SLEDAI score and serum CRP (r =0.396,P ＜ 0.01 ; r =0.351,P ＜ 0.01),and it was not related with ESR or dsDNA (r =0.111,P=0.09;r =0.078,P =0.23).Conclusion The level of serum ferritin could reflect the disease activity of patients with SLE,and it might be used as a new biomarker for disease activity of patients with systemic lupus erythematosus.

Objective To evaluate the serum concentrations of estradiol and testosterone in hyperuricemia patients and possible correlations of the two factors and uric acid (UA) in hyperuricemia (HUA) patients.Methods This was a case control study.We involved 90 hyperuricemia patients,103 healthy controls.Estradiol,testosterone,UA,serum glucose,lipid profile,creatinine and body mass index (BMI) were estimated in two groups.The statistical analysis of the data were performed using SPSS version 19.0 software.The estradiol,testosterone,UA,serum glucose,lipid profile,serum creatinine and BMI levels between the two groups were compared using the Student's t-test.Pearson correlation test was used to assess the correlation between serum UA levels and these indexes.Results Serum levels of estradiol in healthy subjects and hyperuricemia patients were 82.2 (55.6-108.8) pmol/L,65.8 (36.6-95.0) pmol/L respectively ;the serum levels of UA were 300.8(207.2-394.4) μmol/L,426.9(370.1-483.7) μmol/L,respectively.The levels of estradiol were higher in the healthy control group than those in the hyperuricemia group.There were significant differences of estradiol levels between these two groups (P ＜ 0.05).Estradiol was negatively correlated with UA (r =-0.319,P ＜ 0.01),so was testosterone (r =-0.312,P ＜ 0.01).Conclusion The findings of the present study suggest that in hyperuricemia patients,there are associations between estradiol and UA and the levels of serum estradiol might be used as biomarkers in hyperuricemia.

Objective Experimental study on effects of Yangxinshi tablets in the prevention and treatment of chronic is-chemic heart failure and acute myocardial ischemia reperfusion injury in order to provide a theoretical basis for its clinical appli-cation .Method Rat model of chronic ischemic heart failure and acute ischemia reperfusion mouse model of myocardial infarc-tion were constructed through coronary artery ligation .Those animals were randomized to sham operation group ,model group , positive drug group ,high Yangxinshi tablet dose group ,middle Yangxinshi tablet dose group ,low Yangxinshi tablet dose group .Rat electrocardiogram ,ultrasound ,ACE ,ACD ,TNF-a ,mouse myocardial infarction area ,CK ,LDH ,SOD and relat-ed factors were recorded during pre-dosing and post-dosing after modeling .Morphological changes were observed with cardiac pathological section .Results The biochemical indicators in model group were significantly different from sham operation group with statistical significance (P<0 .001) .Compared to the model group ,positive drug group and high Yangxinshi tablet dose group exhibited biochemical differences with statistical significance (P<0 .001 ,P<0 .01 ,P<0 .05) ,significant myocardial in-farction area reduction (P<0 .01 ,P<0 .05) ,improvement in myocardial cells and reduction in the cell infiltration .Conclusion Yangxinshi tablets have preventive and therapeutic effects on chronic ischemic heart failure and acute ischemia reperfusion in-jury .

Liver cirrhosis is an end-stage fibrotic disease of chronic liver injury induced by various factors. Many studies have shown that gut microbiota (GM) are involved in the development and progression of liver cirrhosis and related complications, such as portal hypertension, hepatic encephalopathy, spontaneous bacterial peritonitis, and hepatocellular carcinoma. In addition, impaired intestinal barrier, compromised liver immunity, and bacterial translocation play a critical role in the progression of liver cirrhosis. This article summarizes the role of GM in liver cirrhosis and related complications and research advances and problems in GM as a therapeutic target, in order to provide new thoughts for clinical diagnosis and treatment.

OBJECTIVETo investigate the possibility of multi-unit ribozymes to purge bone marrow of chronic myelocytic leukemia (CML), its in vitro cleavage ability and the reversal effect on CML cell's malignant phenotype.

METHODSAs bcr-abl fusion gene plays an important role in CML pathology, three single-unit ribozymes were designed and synthesized in 44 base pairs near the fusion point, two enzyme cleavage sites on bcr gene and one on abl gene. Multi-unit ribozymes' in vitro transcription and retroviral vector through gene recombination were constructed. Then, its in vitro cleavage ability was tested and the retroviral vector was transfected into K562 cell. Through MTT assay, the incorporation rate of (3)H-TdR, RT-PCR, Southern and Northern blot hybridization, flow cytometry, transmission and scanning electron microscopy were used to study the effect of multi-unit ribozymes on CML cell proliferation, cell structure, cell cycle and the induction of apoptosis.

RESULTSMulti-unit ribozymes had in vitro cleavage efficiency of 70.8%. After the transfection of multi-unit ribozymes retroviral vector into K562 cell, cell proliferation and DNA synthesis were greatly reduced with an inhibition rate of about 50% after 96 hours of transfection. Multi-unit ribozymes could cleave K562 cell's RNA with a reduction rate about one 1 000 th of the original. By flow cytometry (FCM), 18.4% cells underwent apoptosis after 72 hours transfection with most of the cells blocked in the G phase. Here, the ratio in S phase was lowered by 41.9%. Under transmission and scanning electron microscope, compaction of nuclear chromation and apoptosis bodies were observed in the transfected cells.

CONCLUSIONMulti-unit ribozymes possess high cleavage ability in vitro. The ribozymes, whose retroviral vector being transfected into CML cell, are able to express a lasting ability to cleave the fusion gene, induce apoptosis, reduce cell proliferation, revert the malignant phenotype. It is possible to make use of multi-unit ribozymes to purge CML bone marrow. Therefore, multi-unit ribozymes may very well be valuable in the gene therapy of CML.

Apoptosis ; Cell Division ; drug effects ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Humans ; K562 Cells ; RNA, Catalytic ; metabolism ; pharmacology

Objective:To investigate the relationship between the E2 and E4 alleles of apolipoprotein E (apoE) gene and myocardial infarction (MI) in type 2 diabetes Mellitus (T2DM) patients, and to explore the relationship between apoE polymorphism and blood lipid metabolism.Methods:This case control study was conducted from August 2016 to March 2020 in China-Japan Friendship Hospital, 3 459 inpatients with T2DM were included including 3 044 patients without MI (T2DM group) and 415 patients with MI (T2DM+MI group). Real time fluorescent quantitative PCR was used to detect apoE polymorphism. Automatic biochemical analyzer was used to detect lipid levels. Logistic regression analyses were performed to determine the association of apoE with risk of MI in patients with T2DM.Results:(1) The frequency of E4 allele in T2DM+MI group (12.29%, 102/830) was significantly higher than in T2DM group (9.13%,556/6 088), while the frequency of E2 allele in T2DM+MI group (7.35%,61/830) was significantly lower than that in T2DM group (8.21%,500/6 088), P=0.012. Logistic regression analyses showed that E4 allele carrier (E3/E4+E4/E4) faced a higher risk for MI in T2DM patients ( OR=1.48, 95% CI 1.14-1.92, P=0.003), while E2 allele carrier(E2/E3+E2/E2)did not face a higher risk of MI in T2DM patients ( OR=0.88, P=0.642). (2) The levels of apoE polymorphism and blood lipid: The levels of TC, LDL-C and apoB increased in the order of E4 allele, wild type and E2 allele ( P<0.05). The levels of HDL-C, apoA1 and apoE decreased in the order of E4 allele, Wild type and E2 allele ( P<0.05). Conclusion:The E4 allele is a risk factor for MI in T2DM patients, and apoE polymorphism can affect blood lipid level in this patent cohort.

【Objective】 To investigate the risk factors of transfusion-related circulating overload (TACO) in hospitalized patients and to analyze its impact on clinical outcome. 【Methods】 The clinical data of 295 patients with blood transfusion admitted to our hospital from June 2020 to June 2022 were retrospectively analyzed. The patients were divided into TACO group (n=23) and control group (n=272) according to the incidence of TACO. The risk factors of TACO were analyzed by Logistic regression, and the differences of hospital stay and mortality between the TACO group and the control group were compared. 【Results】 TACO occurred in 23 of 295 patients, accounting for 7.80% of all transfusion reactions. The incidence of TACO in different transfusion components was different. Elder age, history of heart failure, history of chronic kidney disease, large mean blood transfusion volume, positive fluid balance [OR(95%CI)): 2.022 (1.212-3.372), 1.917(1.258-2.922), 1.719 (1.155-2.560), 2.252 (1.256- 4.039), 2.221 (1.358-3.633)] were the main risk factors for TACO (P<0.05). 【Conclusion】 Elder age, history of heart failure, history of chronic kidney disease, large blood transfusion volume and positive fluid balance were risk factors for TACO, and TACO was associated with increased length of stay and mortality during hospitalization.