1.Reconstruction of adenovirus vector contained canstatin gene and hTERT gene core promoter and its anti-tumor effect in human ovarian cancer
Baoju ZHU ; Yuhuan QIAO ; Yin LI
Chinese Journal of Obstetrics and Gynecology 2009;44(3):214-218
Objective To study anti-tumor effects on human ovarian cancer xenografted tumors in mice by constructing adenoviral expression vector containing canstatin gene and hTERT gene core promoter (AdhTERT-Can). Methods AdhTERT-Can vector was constructed and identified by means of enzyme cutting, electrophoresis and sequencing. Then, transfected into HO8910PM cells by means of lipofectamine and confirmed by green fluorescence protein(GFP) expression under laser confocus microscope. The mRNA expression of canstatin gene was tested by RT-PCR. Human ovarian cancer xenografted tumor models in nude mice were established and randomly divided into AdhTERT-Can group received viral supematant solution of AdhTERT-Can by tail vein injection, Ad-Can groups received viral supernatant solution of Ad-Can by tail vein injection, and control groups received phosphate buffer solution (PBS). The volume of tumors were measured and compared in each group to evaluated the anti-tumor efficacy. Results All the constructed vectors of AdhTERT-Can were verified by enzymed digestion. There were green fluorescence from 60% of HO8910PM cells transfected by AdhTERT-Can under laser confocus microscope, and the mRNA expression of canstafin gene in HO8910PM cells were also verified by RT-PCR The growth of tumor in AdhTERT-Can group was significantly inhibited compared with those in Ad-Can groups and control groups from the 8th day (P <0.01 ). However, there were not significant difference between Ad-Can group and PBS group (P> 0.05). On the 30th day, the tumors showed liquefaction necrosis and cystic degeneration in each group, especially in AdhTERT-Can group. Conclusions The recombinant adenovirus vector of AdhTERT-Can has been constructed successfully and could steady express in ovarian cancer cell lines HO8910PM. The results shown that it could inhibit significantly the growth of human ovarian cancer xenografted tumors in mice and shown to be the target of gene therapy.
2.The recent advances in the host targets of anti-influenza drugs.
Linlin MA ; Jiandong JIANG ; Yuhuan LI
Acta Pharmaceutica Sinica 2014;49(12):1631-8
The challenge of the emergence of drug-resistant influenza strains, which is caused by wide spread utilization of direct-acting antivirals (DAAs), accelerates the research and exploration towards host targeted agents. In contrast to DAAs targeting viral replication components, host targeted agents, which regulate host factors and pathways linked to viral replication, can interfere the replication of influenza. Additionally, the innate immune system is activated by influenza during the early stage of infection, so manipulating the innate immune response may prevent the viral infection. However, the excessive inflammatory response induced at the late phase of influenza infection would lead to severe tissue injures. Thus, it is very important to explore drugs with anti-inflammatory actions to suppress these immune imbalances and tissue injures. Here we overview the current progresses about host targets related to anti-influenza drugs.
3.Establishment of EV71 animal models with 2-week-old BALB/c mice.
Huiqiang WANG ; Jiandong JIANG ; Yuhuan LI
Acta Pharmaceutica Sinica 2013;48(3):343-6
Animal model is very important for anti-EV71 (enterovirus 71) drug and vaccine development. 1-day-old suckling EV71 mouse model is the main in vivo model used in China. 1-day-old suckling EV71 mouse is too small to perform antiviral experiment. And the route of administration and dosage capacity are also restricted. A strong virulence EV71 virus strain was selected after screening from five EV71 strains with 1-day-old suckling mice. A mouse-adapted EV71 strain with increased virulence in 12-day-old suckling mice, EV71-M5, was generated after five serial passages of the parental EV71 strain in mice. Virus titers of EV71 infected mice heart, liver, spleen, lung, kidney, small intestine, brain and muscle tissue were determined by cytopathic effect (CPE) assay. The virus used in this model is the first isolated EV71 strain in China. And 2-week-old suckling mice were used in this model. This is a supplement for the EV71 animal model in China. Establishment of this EV71 model will provide an attractive platform for anti-EV71 vaccine and drug development.
4.Comparison of percutaneous suturing with sponge forceps and open suturing for repair of acute closed Achilles ruptures
Yuhuan QIN ; Jianyang LI ; Xianteng YANG ; Senlei LI ; Li SUN
Chinese Journal of Orthopaedic Trauma 2017;19(6):500-504
Objective To compare percutaneous suturing with sponge forceps and open suturing for repair of acute closed Achilles ruptures.Methods From March 2011 to June 2014,40 patients with acute closed Achilles rupture were enrolled in this study and randomized into 2 equal groups (n =20) which were subjected to percutaneous suturing with sponge forceps or open suturing.The operative time,incisive length,blood loss,wound healing,hospitalization time and complications were compared between the 2 groups.Their ankle functions were evaluated at postoperative 3,6,and 12 months according to American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot scores.Results The operative time (median,60.0 min),hospitalization time (median,8.5 d) and incisive length (median,8.0 cm) in the open suturing group were significantly greater than those in the sponge forceps group (32.5 min,7.0 d and 1.5 cm) (P < 0.05).The blood loss (15.1 ± 13.7 mL) in the sponge forceps group was significantly less than in the open suturing group (54.2 ± 33.9 mL) (P < 0.05).The open suturing group received an average follow-up of 16.8 months (from 12 to 23 months),witnessed superficial skin infection in 3 cases,and obtained AOFAS scores of 91.2 ± 5.1,95.2 ± 2.8 and 97.8 ± 1.6 at 3,6,and 12 months postoperatively.The sponge forceps group received an average follow-up of 15.6 months (from 12 to 24 months),witnessed wound healing in all cases,and obtained AOFAS scores of 90.6 ±5.3,94.I ±2.6 and 97.0 ±2.6 at 3,6,and 12 months postoperatively.The sponge forceps group obtained insignificantly lower AOFAS scores than the open suturing group (P > 0.05).No injury to the sural nerve or tendon rerupture happened in either group.Conclusions In treatment of acute closed Achilles ruptures,compared with open suturing,percutaneous suturing with sponge forceps may be relatively easier without using special equipment,and additionally,may lead to lower incidence of incision complications and better short-term efficacy.
5.In vitro anti-influenza virus activity of 10 traditional Chinese medicines.
Weiying HE ; Rongmei GAO ; Xingqiong LI ; Jiandong JIANG ; Yuhuan LI
Acta Pharmaceutica Sinica 2010;45(3):395-8
Influenza virus is a virus causing upper respiratory tract infection disease with high morbidity and mortality. China is considered as an area with high rate of influenza morbidity. Prevention and treatment of influenza currently rely on vaccines and antiviral agents in the world. In addition, traditional Chinese medicines also have been used in clinical for influenza therapy. In vitro anti-influenza virus activities of 10 traditional Chinese medicines were studied by cytopathic effect (CPE). Qingre Jiedu oral liquid (factory H) had strong antiviral activity against influenza virus A/Guangdong Luohu/219/2006 (H1N1); Yinhuang oral liquid had strong antiviral activity against influenza virus A/Hanfang/359/95 and A/Yuefang/243/72 (H3N2). Qingkailing oral liquid (factory G) had strong antiviral activity against influenza virus A/Jifang/15/90 (H3N2). Qingre Jiedu oral liquid (factory H) had strong antiviral activity against influenza virus A/Jifang/15/90, A/Yuefang/243/72 (H3N2) and virus B.
6.In vitro and in vivo anti-influenza virus activity of ribavirin injection.
Rongmei GAO ; Xingqiong LI ; Weiying HE ; Jiandong JIANG ; Yuhuan LI
Acta Pharmaceutica Sinica 2010;45(3):403-7
Ribavirin is a broad-spectrum inhibitor against several unrelated DNA or RNA viruses in vitro and in vivo. In this paper the in vitro and in vivo study of anti-influenza virus activity of ribavirin (RBV) injection had been reported. The in vitro antiviral activity of ribavirin injection against influenza virus A and B was studied by CPE. The in vivo protective action of ribavirin injection against influenza A/FM/1/47(H1N1) mouse adapted strain infected mouse was studied with mouse model. The results showed ribavirin injection has strong inhibitory activity against 7 virus strains tested in vitro. Ribavirin injection could significantly increase virus infected mouse survival rate and survival days and improve lung pathogen and lung index.
7.Correlation of Laryngopharyngeal Reflux and Voice Disorders
Sen YAN ; Wen XU ; Yun LI ; Li ZHENG ; Yuhuan ZHANG
Journal of Audiology and Speech Pathology 2014;(1):34-38
Objective To study the correlation between laryngopharyngeal reflux (LPR) and voice disorders . Methods One hundred and three patients with reflux -related symptoms were recruited .The patients were evalu-ated with reflux symptom index (RSI) ,reflux finding score (RFS) evaluation and 24-hour dural-probe pH moni-toring .Eighty -nine cases with voice disorders were divided into 5 groups :vocal process granuloma (n=18) chron-ic pharyngolaryngitis (n=19) ,vocal polyps (n=15) ,vocal fold leukoplakia (n=21) and Reinke's edema (n=16) . The other 14 patients without voice disorders were the control subjects .Results According 24-hour dural -probe pH monitoring assessments ,48 .3% of the patients with voice disorders showed LPR positive .The positive rate in the vocal fold leukoplakia group (71 .4% ) and Reinke's edema group (75 .0% ) were significantly higher than the control group (35 .7% ) .RSI and RFS of the patients with Reinke's edema were both significantly higher than the control group(P=0 .020 ,P=0 .009) .RSI of the patients with chronic pharyngolaryngitis were significantly higher than the control group (P=0 .019) .The acid reflux episodes ,acid reflux time which except in the supine position of the vocal fold leukoplakia group were significantly higher than the control group .The acid reflux episodes ,acid ex-posure and acid reflux time which all except in the supine position of the Reinke's edema group were significantly higher than the control group .Conclusion The correlations between Reinke's edema ,vocal fold leukoplakia and LPR were stronger .In these two groups ,the acid reflux episodes were higher and acid reflux times were longer .
8.A comparative study of escitalopram angumentation with psychoanalysis in the treatment of refractory depression
Xiangjun MENG ; Zhenjing LIU ; Yuhuan LI ; Yuyan JIANG ; Zhonghua SU
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(10):921-922
Objective To investigate the efficacy of escitalopram combined with psychoanalysis in the treatment of refractory depression. Methods A total of 63 patients were randomly divided into escitalopram group ( n = 32) and escitalopram combined with psychoanalysis group( n = 31 ). All patients were evaluated with Hamilton depression Rating Scale(HAMD). Results After treatment,the scores of HAMD in two groups were both significantly lower than those before treatment. In the 8th ( ( HAMD ( 17.35 ± 2.98 ) ), 12th ( ( HAMD (9. 26 ±3.46) )weekend of treatment, the scores of HAMD in study group were significanlly lower than those in control group(8 th:21.97 ± 3.26; 12 th: 15.28 ± 3. 18 ). There were no significant differences in side effects between study group and control group. Conclusion Escitalopram angumented with psychoanalysis takes effects better than escitalopram single and doesn't increase side effects in the treatment of refractory depression.
9.Enzyme kinetics of psoralen and isopsoralen in rat and human liver microsomes
Haiying YANG ; Yuhuan ZHONG ; Lin CHEN ; Hua LI ; Xiaomei ZHUANG
Chinese Journal of Pharmacology and Toxicology 2015;(6):924-930
OBJECTIVE To investigate and compare the enzyme kinetic characters of psoralen (PRN)and isopsoralen(IPRN)in rat and human liver microsomes. METHODS PRN and IPRN in liver microsomes incubates were determined using LC-MS/MS. The enzyme kinetic and metabolic stability of PRN and IPRN were investigated by employing the optimized rat and human liver microsomes incubations. The Vmax and Km values were calculated using the nonlinear regression method. RESULTS The quanti?tative method showed good linearity within the range of 0.1-50.0 μmol · L-1 and was suitable for the assay in biological samples. The in vitro elimination was linear with the substrate concentrations lower than 1 μmol,the protein concentration within 0.5 g · L-1,and the incubation time within 40 min. The t1/2 values of PRN and IPRN in rat and human liver microsomes were 74.5,95.0,74.5 and 173.3 min, respectively. The Vmax values of PRN in rat and human liver microsomes were(1.140±0.080)μmol·min-1·g-1 protein,(0.620±0.060)μmol·min-1·g-1 protein,while Km values of PRN in rat and human liver microsomes were (12.9 ± 0.3)μmol · L- 1,(7.4 ± 1.3)μmol · L- 1,respectively. The Vmax values of IPRN in rat and human liver microsomes were(0.251±0.012)and(0.103±0.014)μmol·min-1·g-1 protein,while Km values of IPRN in rat and human liver microsomes were (3.0 ± 0.4)μmol · L-1,(3.4 ± 0.7)μmol · L-1,respectively. CONCLUSION The enzyme kinetic characters and metabolic stability of PRN and IPRN show species and chemical structures related differences. Interestingly,the metabolic eliminations of PRN and IPRN are similar in rats. However,the metabolic elimination of IPRN in humans involved in CYP enzymes may be much slower than that of PRN.
10.Quality control in maternal serum prenatal screening in the second trimester
Fengxian SHEN ; Li ZHAO ; Shiming LYU ; Yuhuan SHEN
Chinese Journal of Laboratory Medicine 2014;(11):870-875
Objective To analyze the features of quality management of maternal serum prenatal screening for Down syndrome in the second trimester and investigate the methods to establish and improve quality control system.Methods Prenatal screening and clinical data were collected for statistical analysis from 17 prenatal screening centers in Zhejiang Province from 2011 to 2012.The impacts of maternal age , weight, gestational age , specimen collection , the median values of serum markers selection and different analysis software on prenatal screening results for Down syndrome were analyzed respectively .Results Totally 417 347 and 463 846 prenatal screening cases were collected in 2011 and 2012 respectively.The maternal age of the entire province presented as a Gaussian distribution.The age distributions of different screening institutions were inconsistent , while maternal weight distributions were similar.The approach to determine gestational age was significantly different among screening institutions.Median MoM of the samples from the blood collection points was different from those of the main screening center.Median MoM-freeβ-hCG was positive bias , particularly the specimens from the blood collection points.After strengthening the control of temperature and time during samples transportation , Median MoM-free β-hCG of specimens from blood collection points became nearly equal to those of main screening center .Use of local median made the median MoM value closer to 1.0 in the centers with positive or negtive bias of the markers , which had little effect in detection rate and decreased the false positive rate and screening positive rate.Six more positive cases were detected by the Lifecycle software instead of 2T software in 213 followed-up positive cases, which means the Lifecycle software can improve the detection rate.Conclusions The impact of maternal age, weight, gestational age, specimen collection and other factors on screening performance should be taken into consideration when it evaluates the screening quality of different laboratory .The use of local medians is necessary in prenatal screening.Optimization of screening analysis software can reduce false negative rate.