1.The discovery of the interaction between augmenter of liver regeneration and Na~+, K~+-ATPase with yeast two-hybrid system
Minghua TONG ; Siqiang CHEN ; Ruhua YAO ; Xiangpin KONG
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To screen the proteins interacting with human augmenter of liver regeneration (hALR) by yeast two-hybrid system and to study the mechanism of hALR action. METHODS: hALR bait plasmid was constructed by ligating the gene of hALR into pGBKT7, then transformed into yeast AH109. The yeast strain AH109 containing pGBKT7-hALR was mated with yeast Y187 containing human liver cDNA library plasmid. Diploid yeast was plated on SD/-trp-leu-his-ade (QDO) for screening and on QDO containing X-?-gal for further selection.The AD/library inserts were amplified by PCR and the PCR products were characterized by digesting with Sau3AⅠ and HaeⅢ restriction enzyme to eliminate the duplicates. After sequencing, the positive clones were analysed by bioinformatics. RESULTS: Several positive clones were obtaind. The sequencing and analysis shown that one of them is 669 bp DNA fragment encoding ? subunit of Na +,K +-ATPase. The 224 bp 3′terminal DNA fragment is non-encoder region, and the 445 bp 5′terminal DNA encodes C-terminal 147 amino acid residues of Na +, K +-ATPase ? subunit. CONCLUSION: The results of screening proteins using yeast two-hybrid system showed that hALR could interact directly with Na +, K +-ATPase in the yeast cell.
2.A multicenter study of effect of pidotimod on the immune function in children with respiratory tract infection
Yanyuan YANG ; Hua ZHANG ; Zhanling LI ; Xiuhui XU ; Huiyao ZHONG ; Ruhua CHEN
Chinese Journal of Biochemical Pharmaceutics 2014;37(7):132-134,137
Objective To explore the clinical curative effect of pidotimod on patients with respiratory tract infection and effect on immune function. Methods 120 children with recurrent respiratory tract infection in the Third Hospital of Qinhuangdao,the Third Staff Hospital of Baogang Group,the Third Hospital of Wulanchabu were selected,and were divided into two groups according to random number table.60 cases in control group were treated with routine treatment of anti-infection,relieving cough,eliminating phlegm,antipyretic;60 cases in experimental group were treated with pidotimod on the basis of routine treatment,oral with boiled water,0.4g per times,2 times a day,with a course of 60 days.Clinical curative effect after treatment and serum immunoglobulin (IgG,IgA,IgM)levels,T lymphocyte subsets (CD3+,CD4+,CD8+)levels and NK cells relative activities before and after treatment were compared between two groups.Results After treatment,the total effective rate of experimental group (95.00%)was significantly higher than that of control group (81.67%),and the difference was statistically significant (P<0.05);the immune indexes before treatment had no significant difference,and levels of serum immunoglobulin and T lymphocyte subsets were improved,and levels of serum immunoglobulin (IgG,IgA,IgM)and T lymphocyte subsets (CD3+,CD4+,CD8+)of experimental group were more higher than those of control group,and the difference was statistically significant (P<0.05 );relative activity of NK cells in both groups improved after treatment,but relative activity of NK cells in experimental group was significantly higher than that in control group,and the difference was statistically significant (P<0.05 );adverse reactions according minor rashes and anemia were observed in two groups,and there was no significant differece in the incidence of adverse reactions,and ADR was tolerable after symptomatic treatment.Conclusion Pidotimod could significantly improve the clinical curative effect of patients with respiratory tract infections and effectively improve the immune function of patients with recurrent respiratory tract infections with high security,which has a clinical significance.
3.Studies on pharmacokinetics of Musk Protecting Heart Pellets with pH-dependent gradient-release and Musk Protecting Heart Pills
Hongtao SONG ; Tao GUO ; Ruhua ZHANG ; Haiyang HU ; Xian LI ; Xianyin CHEN
Chinese Traditional and Herbal Drugs 1994;0(09):-
Object To investigate the pharmacokinetic parameters of Musk Protecting Heart Pellets with pH-dependent gradient-release (MPHP-pH) and Musk Protecting Heart Pills (MPHP). Methods The cardiac muscle nutritional blood flow in rat was measured as effective index. Results It was one-compartment model when the rat was ig MPHP. The minimal effecting dose was 0.54 mg/kg, the present half-life, the elimination half-life, medicinal effect and the peak time of effective action were 0.53, 1.21, 3.48 and 1.13 h, respectively. The absorption half-life, elimination half-life and peak time of effective dose were 0.23, 1.47 and 0.88 h, respectively. Statistical moment analysis showed that the mean residence time mean residence time (MRT) of effective action were 5.05 h for MPHP-pH and 2.33 h for MPHP, the MRT of effective dose were 7.70 h for MPHP-pH and 3.21 h for MPHP. The relative bioavailability of effective dose for MPHP-pH was 104.03%. Conclusion MPHP has the characteristics of fast absorption, fast eliminationand short effective action time. Whereas MPHP-pH has the characteristics of fast absorption, protonged and relaxed effective action compared with MPHP.
4.Efficacy and safety of single-bolus tenecteplase compared with front-loaded alteplase in Chinese patients with acute myocardial infarction
Feng LIANG ; Dayi HU ; Xubo SHI ; Mingshu GAO ; Jiaping WEI ; Hong ZHAO ; Sanqing JIA ; Hongyu WANG ; Ruhua LIU ; Yundai CHEN ; Yanling LU
Journal of Geriatric Cardiology 2007;4(3):137-141
Background and Objective Previous study showed tenecteplase and alteplaxe were equovalent for 30-day mortality in the treatment of acute myocardial infarction. The purpose of this open-label, randomized, multi-center, angiographic trial was to assess the efficacy and safety of tenecteplase compared with alteplase in Chinese patients with acute myocardial infarction. Methods We recruited patients with acute ST-elevation myocardial infarction presenting within 6 hours of symptom onset from October, 2002 to March,2004, in 5 hospitals in Beijing. After giving informed consent, patients were randomly assigned a single-bolus injection of tenecteplase (30-50 mg according to body weight) or front loaded alteplase (100 mg), and underwent coronary angiography at 90 min after starting the study drug. All patients received aspirin and heparin (target activated partial thromboplastin time 50-70 s). The primary efficacy end point was the rate of TIMI grade 3 flow at 90 minutes. Other efficacy end points included TIMI grade 2/3 flow at 90 minutes. Safety end points included all stroke, intracranial hemorrhage (ICH), moderate/severe hemorrhage (except for ICH), all-cause mortality at 30-days, and major non-fatal cardiac events at 30 days. Results Overall 110 patients were eligible for statistical analysis, with 58 patients assigned to receive tenecteplase and 52 patients to alteplase. Tenecteplase produced a rate of TIMI grade 3 flow at 90 minutes after the start of thrombolysis(68.4%) similar to that of alteplase (66.7%, P=1.0); the rates of TIMI grade 2 or 3 were similar for patients treated with tenecteplase versus alteplase (89.5% versus 80.4%, respectively, P=0.278). At 30 days, rates for all strokes were similar for the two groups (5.17% for tenecteplase and 1.92% for alteplase, P=0.62); rates of ICH were 3.45% and 1.92% (tenecteplase and rt-PA,P=1.00) respectively. The rate of moderate/severe hemorrhage was 8.62% with tenecteplase and 5.77% with alteplase (P=0.72); total mortality was almost identical in the two groups (13.8% versus 9.6%, respectively, P=0.565) while the rates of non-fatal cardiac complications were 10.35% and 11.54% (tenecteplase and alteplase, P=1.0). Conclusions The efficacy of a single-bolus, weightadjusted tenecteplase fibrinolytic regimen is equivalent to front-loaded alteplase in terms of the rates of TIMI grade 3 flow, and TIMI 2 or 3 flow, but the 30-day mortality and ICH in both groups was so high that the use of tenecteplase is not permitted in China. These negative safety results might be due to the high rate of percutaneous coronary intervention (PCI) and high dose of bolus heparin and suboptimal concomitant medical therapy during hospitalization, so further studies are needed to confirm the safety for tenecteplase in Chinese patients.
5.Effect of small nuclear ribonucleoprotein polypeptide A on the malignant biological behavior of hepatocellular carcinoma cells and its mechanism
YAO Menglin ; WANG Ruhua ; CUI Xiaomeng ; CHEN Yifei ; GUO Dan ; HE Shuixiang ; LI Yarui
Chinese Journal of Cancer Biotherapy 2023;30(12):1074-1081
[摘 要] 目的:探究小核核糖核蛋白多肽A(SNRPA)在肝细胞癌(HCC)组织和细胞中的表达及其调控HCC细胞HepG2和Hep3B恶性生物学行为的作用及其机制。方法: 数据库分析SNRPA在泛癌组织中的表达及其与病理分期、HCC患者预后的相关性。常规培养HepG2和Hep3B细胞,将si-NC,si-SNRPA#1、si-SNRPA#2转染HepG2和Hep3B细胞,实验分为si-NC组、si-SNRPA#1组和si-SNRPA#2组;将SNRPA-vector和SNRPA-oe载体转染LO2细胞,分为SNRPA-vector组和SNRPA-oe组。qPCR法检测正常肝细胞和肝癌细胞以及转染各组HepG2和Hep3B细胞中SNRPA mRNA的表达,MTT法、Transwell法和WB法分别检测转染后各组HepG2和Hep3B细胞的增殖、迁移和侵袭能力以及EMT相关蛋白表达的变化。结果: 数据库分析显示,SNRPA mRNA在多数肿瘤组织中均呈高表达(均P<0.001)且与病理分期有关联(P<0.05或P<0.01)。SNRPA在HCC组织和细胞中均呈高表达(P<0.05或P<0.01),且与HCC患者的预后有关联(P<0.01)。敲减SNRPA表达明显抑制HepG2和Hep3B细胞增殖(P<0.05或P<0.01)而过表达SNRPA则能促进LO2细胞增殖(P<0.01),敲减SNRPA表达明显抑制HepG2和Hep3B细胞的迁移和侵袭能力(均P<0.01),明显促进E-cadherin的表达上调(P<0.01),而抑制N-cadherin、vimentin的表达(P<0.01)。结论: SNRPA在HCC组织及细胞中呈明显高表达,其可能通过调控上皮间质转化(EMT)进程进而促进HepG2和Hep3B细胞的增殖、迁移和侵袭。
6.Role of lncRNA SNHG5 in the migration and invasion of hypoxia-induced hyhepa‐ tocellular carcinoma cells
LI Yarui ; GUO Dan ; CHEN Yifei ; WANG Ruhua ; LU Guifang ; REN Mudan ; LU Xinlan ; HE Shuixiang
Chinese Journal of Cancer Biotherapy 2020;27(9):978-983
[Abstract] Objective: To explore the regulatory effect of long non-coding RNA (lncRNA) SNHG5 on invasion and migration of
hypoxia-induced hepatocellular carcinoma (HCC) cells. Methods: A total of 20 pairs of cancer and para-cancerous tissue specimens
resected from HCC patients in the First Affiliated Hospital of Xi'an Jiaotong University from January 2017 to June 2018, and human
HCC cell lines (HepG2, MHCC-97L, MHCC-97H , Huh7) as well as immortalized human liver LO2 cells were collected for this study.
Bioinformatics methods were used to analyze the binding sites between hypoxia-inducible factor 1α (HIF-1α) and SNHG5. pCMVHIF-1α and shRNA-SNHG5 (sh-SNHG5) plasmids were transfected into HCC cells, respectively. qPCR was used to detect the expres‐
sion level of SNHG5 in HCC tissues and hypoxia-induced HCC cells. Western botting was used to detect the expression level of HIF-1α
protein in HCC cells, and Transwell chamber method was used to detect the migration and invasion ability of HCC cells after SNHG5 si‐
lence under normoxia and hypoxia condition. Results: Compared with para-cancerous tissues and immortalized human liver LO2 cells,
the expression of SNHG5 was significantly up-regulated in HCC tissues and cell lines (all P<0.01). Hypoxia promoted the expression
level of SNHG5 in HCC cells, and its mechanism might be related to the combination of hypoxia-activated HIF-1α and SNHG5
promoter to promote its transcription. Hypoxia promoted the invasion and migration ability of HepG2 and MHCC-97L cells (all P<
0.01), but knockdown of SNHG5 significantly inhibited the invasion and migration ability of HepG2 and MHCC-97L cells under hy‐
poxic conditions (all P<0.01). Conclusion: SNHG5 is highly expressed in HCC tissues and cell lines and plays an important role in the
invasion and migration of HCC cells induced by hypoxia.