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Thyroid hormones(TH), one of the human′s essential hormones, play a crucial role in the cardiovascular system.Studies have shown that hypothermia, blood dilution, vascular endothelial injury, ischemia-reperfusion, and inflammatory factor release during cardiopulmonary bypass may cause thyroid dysfunction, leading to the euthyroid sick syndrome(ESS). There is a close correlation between ESS and postoperative low cardiac output and elevated systemic vascular resistance, which seriously affects the prognosis of pediatric patients.Studies have shown that perioperative supplementation of thyroid hormones can reduce ESS levels, especially among pediatric patients and those children with complex congenital heart disease have apparent clinical advantages.However, the results from different clinical studies varied, and currently, thyroid hormone replacement therapy is under debate.This review examines the available literature on the clinical effects of thyroid hormone on the cardiovascular system and the relationship between ESS and cardiopulmonary bypass.The clinical evidence of the treatment of ESS is gathered and discussed with an intent to find a gap for further research.
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Objective@#To explore the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection who received entecavir alone or in combination with anluohuaxianwan for 78 weeks.@*Methods@#Patients with chronic HBV infection were randomly treated with entecavir alone or in combination with anluohuaxian for 78 weeks. Ishak fibrosis score was used for blind interpretation of liver biopsy specimens. The improvement in liver fibrosis condition before and after the treatment was compared. Student's t test and non-parametric test (Mann-Whitney U-Test and Kruskal-Wallis test) were used to analyze the measurement data. The categorical variables were analyzed by Chi-square test method and Spearman’s rank correlation coefficient was used to test bivariate associations.@*Results@#Liver fibrosis improvement rate after 78 weeks of treatment was 36.53% (80/219) and the progression rate was 23.29% (51/219). The improvement of liver fibrosis was associated to the degree of baseline fibrosis and treatment methods (P < 0.05). The improvement rate of hepatic fibrosis in patients treated with anluohuaxianwan combined with entecavir at baseline F < 3 (54.74%, 52/95) was significantly higher than that in patients treated only with entecavir (33.33%, 16/48), P = 0.016 and the progression rate of hepatic fibrosis (13.68%, 13/95) was lower than that in patients treated alone (18.75%, 9/48), P = 0.466. In patients with baseline F < 3, the proportion of patients with improved and stable liver fibrosis in the combined treatment group (68.1%, 32/47) was higher than that in the treatment group alone (51.7%, 15/29).@*Conclusion@#Combined anluohuaxianwan and entecavir treatment can significantly improve the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection. Furthermore, it has the tendency to improve the stability rate and reduce the rate of progression of liver fibrosis.
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Objective@#To evaluate the efficacy and safety of sofosbuvir (Nanjing Zhengda Tianqing Pharmaceutical Co., Ltd.) combined with ribavirin in patients with genotype 2 chronic hepatitis C virus infection.@*Methods@#Treatment-naïve or treatment experienced genotype 2 chronic hepatitis C patients from sixteen research centers of China were screened. All subjects received once-daily dose of sofosbuvir (400 mg) combined with ribavirin (body weight < 75 kg, 1 000 mg/day, 400 mg in the morning and 600 mg in the evening; body weight > 75 kg, 1 200 mg/d, 600 mg in the morning and 600 mg in the evening) for 12 weeks. Patients were followed-up for a period of 12 weeks after discontinuation of treatment. Continuous variables were expressed as mean ± standard deviation. The proportion of subjects with virologic response at different follow-up time points and 95% confidence intervals were estimated by maximum likelihood ratio and Clopper-Pearson interval.@*Results@#132 cases with genotype 2 chronic hepatitis C virus infection from sixteen research centers of China were included, 12 cases of whom were associated with cirrhosis, and the remaining 120 cases were not associated with cirrhosis. One hundred and thirty-one cases completed the study, and one patient lost to follow-up at week 4 after the end of treatment. The sustained virological response rate was 96.2% (95% confidence interval: 92.37% - 99.16%) after 12 weeks of drug withdrawal. Virological relapse occurred in four cases. Of the 132 subjects enrolled in the study, 119 (90.2%) reported 617 adverse events during treatment, of which 359 (76.5%) were TEAE related to sofosbuvir and/or ribavirin. There were nine TEAEs of grade 3 and above, and six cases (4.5%) of them had six severe adverse events. Only one serious adverse event was associated with sofosbuvir and ribavirin (unstable angina pectoris). There were no adverse events leading to drug discontinuation or death.@*Conclusion@#Sofosbuvir combined with ribavirin has a high SVR rate in the treatment of genotype 2 chronic hepatitis C virus infection, and most of the adverse events occurred were mild with acceptable safety profile.
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Objective@#To evaluate the safety and efficacy of ombitasvir/paritaprevir/ritonavir (OBV/PTV/r) 25/150/100 mg once daily combined with dasabuvir 250mg, twice daily in non-cirrhotic Chinese adult patients with newly diagnosed and treated chronic HCV genotype 1b infection. @*Methods@#A randomized, double-blind, placebo-controlled, multicenter phase 3 clinical trial was conducted in mainland China, Korea, and Taiwan.Safety and efficacy of OBV/PTV/r plus DSV administered for 12 weeks were evaluated in a newly diagnosed and treated (interferon alpha /pegylated interferon alpha) and ribavirin non-cirrhotic adults with chronic HCVgenotype 1b infection. Patients randomly received OBV/PTV/r plus DSV for 12 weeks (Group A), or placebo for 12 weeks (Group B) followed by an open-label phase of OBV/PTV/r plus DSV for 12 weeks. Sustained response (SVR12) rate obtained at 12 weeks and (SVR24) 24 weeks after discontinuation of treatment, and the incidence of adverse events and laboratory abnormalities after double-blind and open-label phase treatment were assessed. @*Results@#A total of 410 cases of Chinese patients were included and randomly assigned to group A and B (with 205 cases in each group) in a 1:1 ratio. The rates of SVR12 and SVR24 were 99% (95% CI: 94.8% - 99.8%) in the newly diagnosed patients in group A (205 patients) and the rates of SVR12 and SVR24 were 100% in treated patients (95% CI: 96.3% - 100%). Different baseline characteristics had no effect on SVR12 and SVR24 rates. Most of the adverse events occurred were mild, asymptomatic, and≥ 3 laboratory abnormalities during treatment were rare, including elevation of alanine aminotransferase (2 cases in double-blind stage A group), aspartate aminotransferase (Double-blind stage A (3 cases) and total bilirubin (1 case in open-label phase B group); however, those mild adverse events could be recovered after drug withdrawal or discontinuation. only1 person discontinued drugs due to adverse events (Group B, open-label phase). @*Conclusion@#The 12 weeks treatment course of OBV/PTV/r combined with DSV produced 99% ~ 100% rates of SVR12 and SVR24 in non-cirrhotic Asian adult patients with newly diagnosed and treated chronic HCV genotype 1b infection, and the tolerance and safety were good.
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Objective@#To evaluate the efficacy and safety of ombitasvir/paritaprevir/ritonavir (OBV/PTV/r) 25/150/100 mg once daily and dasabuvir (DSV) 250 mg twice daily combined with ribavirin in adult patients of Mainland China with chronic HCV genotype 1b infection and compensated cirrhosis. @*Methods@#An open-label, multicenter, phase 3 clinical trial study was conducted in mainland China, Taiwan, and South Korea. Adult patients with compensated cirrhosis (Metavir score =F4) who were newly diagnosed and treated for hepatitis C virus genotype 1b infection with ombitasvir/paritaprevir/ritonavir and dasabuvir combined with ribavirin for 12 weeks were included. Assessed SVR rate of patients obtained at 12 and 24 weeks after drug withdrawal. Efficacy and safety were evaluated in patients who received at least one time study drugs. @*Results@#A total of 63 patients from mainland China were enrolled, 62 of whom (98.4%) had a baseline Child-Pugh score of 5 points. The overall rate of SVR12 and SVR24 in patients was 100% (95% CI: 94.3% to 100.0%). Most of the adverse events that occurred were mild. The incidence of common (≥10%) adverse events and laboratory abnormalities included elevated total bilirubin (36.5%), weakness (19.0%), elevated unconjugated bilirubin (19.0%) and conjugated bilirubin (17.5%), and anemia (14.3%). Three cases (4.8%) of patients experienced Grade ≥ 3 adverse events that were considered by the investigators to be unrelated to the study drug. None patients had adverse events leading to premature drug withdrawal. @*Conclusion@#Mainland Chinese patients with chronic HCV genotype 1b infection and compensated cirrhosis who were treated with OBV/PTV/r plus DSV combined with RBV for 12 weeks achieved 100 % SVR at 12 and 24 weeks after drug withdrawal. Tolerability and safety were good, and majority of adverse events were mild.
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Objective@#To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control.@*Methods@#This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data.@*Results@#A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events.@*Conclusion@#In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.
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Objective@#To establish the model of liver fibrosis based on noninvasive indices, and to investigate the diagnostic value of this model.@*Methods@#A total of 838 patients with chronic hepatitis B (CHB) who underwent liver biopsy in our hospital from March 2003 to October 2013 were selected, and the results of blood tests and B-ultrasound were collected. The correlation between these indices and liver fibrosis stage was analyzed. A logistic regression analysis was performed to establish a predictive model, and the value of this model was examined in validation group. The t-test, Mann-Whitney U non-parametric test, and chi-square test were used for data analysis. A Spearman rank correlation analysis was used for bivariate correlation analysis, and a dichotomous logistic stepwise regression analysis was used for multivariate analysis.@*Results@#In the model group, a model (FV) consisting of age, platelet count (PLT), γ-glutamyl transferase (GGT), albumin/globulin ratio (A/G), and splenic square area (SSA) was established. The areas under the receiver operating characteristic curve (AUROCs) of the model FV were 0.892, 0.910, and 0.915, respectively, in diagnosing significant liver fibrosis (S2-4), progressive liver fibrosis (S3-4), and early-stage liver cirrhosis (S4), with sensitivities of 77.6%, 83.7%, and 86.0%, respectively, specificities of 89.7%, 84.5%, and 83.7%, respectively, and accuracy of 82.1%, 84.2%, and 84.2%, respectively. There were no significant differences in AUROCs between the validation group and the model group (Z = 0.360, 0.885, and 0.046, all P > 0.05). In all patients, FV had significantly higher AUROCs in the diagnosis of liver fibrosis than FIB4 index and S index (Z = 4.569/3.423, 5.640/4.709, and 4.652/4.439, all P < 0.05). With < 0.374 and ≥ 0.577 as the cut-off values for the exclusion and diagnosis of significant liver fibrosis, 61.1% (512/838) of all patients could avoid liver biopsy, and the accuracy was 92.6% (474/512).@*Conclusion@#The noninvasive model based on age, PLT, GGT, A/G, and SSA can accurately predict liver fibrosis degree in patients with CHB with good reproducibility; therefore, it can be used for dynamic monitoring of liver fibrosis degree in clinical practice.
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Objective:To explore the clinical characteristics, mechanism and countermeasures of drug-induced second thrombocy-tosis ( ST) in order to ensure safe medication. Methods:Drug-induced ST published during 1996 and 2016 in domestic and overseas medical journals was collected and statistically analyzed in respects of age, gender, kind and distribution of drug-induced ST, treatment and outcome of ST. Results:Totally 198 cases were collected, which involved 30 different drugs of eight types mainly including antitu-mor drugs, blood system drugs and antibiotics. Among the patients, 112 ones (56. 6％) were male and the other 86 ones (43. 4％) were female. Totally 14 patients had embolization with the overall incidence of 7. 0％. Conclusion:Without timely and favorable treat-ment, drug-induced ST may lead to thrombosis. To ensure the safety of medication, platelet level should be monitored regularly during medication.
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Objective To investigate the epidemiological and clinical characteristics of measles in patients of different age groups in Northeast China in 2014.Methods The clinical data of patients with measles in ten hospitals of infectious diseases in Northeast China from January 2014 to June 2014 were collected.Patients were divided into <16 age group and ≥16 age group, and the epidemiology , clinical characteristics, treatment and prognosis of all patients were retrospectively reviewed .SPSS 17.0 was used for data analysis.Results There were 1 401 patients with measles, in which 402 were <16-year old, and 999 were ≥16-year old.Clinical manifestations were mainly maculopapule (100.0%), fever (84.58%), cough (85.80%), Koplik’ s spot (73.23%), pharyngeal hyperemia (71.23%), diarrhea (43.97%), expectoration (40.97%), tears (39.40%) and runny nose (30.55%).The incidences of Koplik’s spot, sputum, pharyngeal hyperemia and diarrhea in <16 age group were lower than those in ≥16 age group (χ2 =8.364, 29.768, 10.953 and 6.701, P<0.05 or <0.01); while the incidence of runny nose was higher than that in ≥16 age group (χ2 =6.703, P<0.05).Abnormalities were found in blood routine examination, C-reactive protein (CRP), liver and kidney function, serum electrolyte levels, myocardial enzymes, and so on.Increasing in WBC, PLT and creatine kinase isoenzyme (CKMB), and decreasing in WBC were observed in 38(9.45%), 122(30.35%), 279(69.40%) and 105(26.12%), patients in <16 age group, which were higher than those in ≥16 age group [45(4.5%), 14(1.40%), 347(34.73%) and 202(20.22%)], and the differences were of statistical significance (χ2 =12.593, 274.033, 139.385 and 5.830, P<0.05 or P<0.01).Increasing in alanine aminotransferase (ALT), CRP, total bilirubin level (TBil), creatine kinase (CK), and decreasing in albumin (Alb), K+, Na+, Cl-were observed in 70(17.41%), 7(1.74%), 38(9.45%), 7(1.74%), and 214(53.23%), 59(14.68%), 45(11.19%), 94(23.38%) patients in <16 age group, which were lower than those in ≥16 age group [668(66.87%), 89(8.91%), 277(27.73%), 714(71.47), and 268(26.83%), 339(33.93%), 642(64.26%), 450 (45.05%)], and the differences were of statistical significance (χ2 =281.230, 23.073, 50.687, 159.740, and 14.674,114.286, 44.268, 271.546, P<0.01).Laryngitis and pneumonia were the most common complications.The incidence of laryngitis in <16 age group was 12.69% (51/402), which was higher than that in ≥16 years group (93/999, 9.31%,χ2 =3.545, P<0.05);while the incidence of spot shadows demonstrated by X-ray in <16 years group ( 72.89%, 121/166 ) was higher than that in ≥16 years group (265/445, 59.55%,χ2 =9.249, P<0.01).Conclusions There are differences in clinical features of measles in patients between <16 age group and ≥16 age group.Basic immunization in children and revaccination in adults should be enhanced to control the epidemics of measles .
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Objective:To investigate the pharmaceutical care methods for the patients with drug-induced liver injury. Methods:The participation process of clinical pharmacists in 3 cases of typical drug-induced liver injury was with detailed introduction and analy-sis in the paper. Results:The case 1 indicated that new drugs probably with drug-induced liver injury should be warned in the process of clinical medication. The case 2 suggested that TDM, as a useful assessment weapon, could be fully used to find the source medi-cines when drug-induced liver injury occurred. The case 3 showed the specific cases, especially the patients with abnormal liver func-tion, should be focused on, the medicines with high liver toxicity should be avoided and the medicines with mild liver toxicity could be chosen. Conclusion:Clinical pharmacists should participate in the clinical practice of drug-induced liver injury with multi-channel and multi-link, and pay attention to the drugs with high risk of liver injury. Meanwhile, clinical pharmacists should perform TDM monito-ring to provide positive evidence for the diagnosis of drug-induced liver injury, and focus on the liver and kidney functions to provide better pharmaceutical care for the patients.
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Objective To investigate the application of ultrasound and blood indicators in diagnosis of early liver cirrhosis (S4) for patients with chronic hepatitis B (CHB).Methods Data of blood and ultrasound examinations of 631 patients with CHB who received liver biopsy in the Sixth People' s Hospital of Shenyang during April 2002 and March 2011 were collected.Logistic regression analysis was performed to determine the factors independently correlated with early liver cirrhosis,and the diagnostic model was established using these indicators.The diagnostic value of the established model was assessed by using area under receiver operating characteristic curve (AUROC) and compared with FIB-4 index,aspartate aminotransferase (AST)/platelet (PLT) ratio index (APRI) and S index.Results Logistic regression analysis indicated that age,PLT,albumin globulin ratio (A/G) and splenic square area (SPS) in ultrasound findings were independently correlated to early liver cirrhosis (Wald =10056,46.236,3.751 and 10.669,P ＜0.01).AUROC of the model based on the above factors in diagnosing early liver cirrhosis was 0.908,which was higher than those of FIB-1 index,APRI index and S index (Z =8.322,4.334 and 4.087,P ＜ 0.05).Taking 0.063 as cut-off value,the sensitivity,specificity,positive predict value and negative predict value of the established model in diagnosis of early liver cirrhosis were 90.1％,77.8％,50.0％ and 97.1％,respectively.Taking ＜0.060 and ≥0.110 as the cut-off values to exclude,and diagnose early liver cirrhosis,69.7％ (440/631) patients could avoid liver biopsy.Conclusion The model based on age,PLT,A/G and SPS can effectively predict early liver cirrhosis,and can reduce liver biopsy.
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Objective To compare the clinical data of patients with chronic hepatitis B virus (HBV) carriers and chronic hepatitis B so that to provide pathological evidence for management of chronic HBV carriers with different alanine aminotransferase (ALT) levels.MethodsLiver biopsies were performed in totally of 292 cases of chronic HBV infection.The subjects were divided into HBV carrier group (G0-G1 and S0-S1) and hepatitis group (G＞ 1 and/or S＞ 1) according to the pathological diagnosis. The relationships between different age subgroups, different ALT level subgroups and pathological diagnosis were analyzed. Meanwhile,other clinical,biochemical,and iconographic indexes which were possibly related to the pathology diagnosis were compared. The multivariate analysis was done by Logistic regression equation (withdrawal method, maximum likelihood method) to definite the independent influencing factors of pathologically diagnosed with chronic HBV carrier.ResultsAmong the 292 patients,140 (47.9％) were pathologically diagnosed with chronic HBV carries and 152 (52.1％)were chronic hepatitis B. There were statistical differences between ≤35 years group and 36-40 years,＞40 years group (x2 =3.936 and 8.534,respectively; P =0.047 and 0.003,respectively). There were statistical difference among patients with ALT＜0.5×upper limit normal (ULN),(0.5-1.0) ×ULN,(1.1-1.5) ×ULN,(1.6-2.0) ×ULN and ＞2.0 × ULN (x2 =55.314,P＜0.01),while there was no significant difference between (1.1-1.5) × ULN and ＞2.0 × ULN (x2 =3.810,P=0.051). Multivariate analysis indicated that course of disease,alcohol consumption,ALT,HBV DNA level and the surface of liver (smooth or not smooth)detected by ultrasonography were independent influencing factors of pathological diagnosis of chronic HBV carriers (OR =0.995,0.224,0.516,1.308 and 0.270,respectively; P=0.005,0.007,0.000,0.025 and 0.001,respectively).ConclusionLiver biopsy is much meaningful in patients with age ＞35 years old and ALT (1-2)× ULN.
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ObjectiveTo investigate the renal protective effect of recombinant lentivirus encoding adiponectin gene on streptozocin-induced early diabetic nephropathy(DN) mice,and to explore its potential mechanism.Methods Forty C57BL/6 mice were randomly divided into normal control group(NC group,n=10),diabetic nephropathy group(DN group,n=10),LentiIRES-EGFP treatment group(DL group,n=10) and Lenti-Acdc-IRES-EGFP treatment group (DA group,n=10).After 8 weeks of recombinant lentivirus injection,kidney to body weight ratio (KW/BW),mean glomerular volume(MGV),fractional mesangial area(FMA),24 h urinary protein excretion(UTP),Scr,BUN,serum albumin and adiponectin were measured.Renal pathological changes were evaluated by electron microscopy.Proliferation of glomendar and tubulointerstitial cells was assessed by immunohistochemistry using PCNA antibody.The phosphorylation of AMP-activated protein kinase(AMPK) and mammalian target of rapamycin protein(mTOR) were detected by Western blotting.Results Adiponectin was successfully over-expressed in STZ-induced DN mice after lentivirus injection.KW/BW,MGV,FMA and UTP were significantly decreased in DA group as compared to DN group and DL group(P＜0.05),but were increased as compared to NC group(P＜0.05).DA group animals had significantly fewer PCNA-positive cells than DN group and DL group(P＜0.01).DA group mice had higher p-AMPK level and lower p-mTOR level as compared to DN group and DL group(P＜0.01).Conclusion Over-expression of adiponectin has beneficial effect on early DN and attenuates aberrant proliferation of renal cells via AMPKmTOR pathway.
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Objective To evaluate the effects of recombinant lentivirus encoding human apM1 gene ( LentiapM1-EGFP) on platelet-derived growth factor (PDGF) induced human mesangial cell (HMC) proliferation and intracellular AMP-activated protein kinase(AMPK) signaling pathway.Methods Protein expression of apM1 in cell culture supernatant of HMC transfected with Lenti-apM1-EGFP was detected by ELISA.The effect of human adiponectin on cell proliferation and cell cycle was assessed by [ 3 H ] thymidine incorporation assay and Flow cytometry respectively.The phosphorylation of AMPK was detected by Western blotting.Results Lenti-apM1-EGFP had no significant toxicity on HMC.The 50 multiplicity of infection (MOI) of the Lenti-apM1-EGFP efficiently infected HMC,and made it stable expression of adiponectin protein ( 149.6 ± 12.8 ) μg/L.PDGF-induced HMCs proliferation was significantly inhibited by adiponectin.When co-treatment with compound C,an AMPK inhibitor,the inhibitory effort was reversed.The phosphorylation level of AMPK was increased in HMC transfected Lenti-apM1-EGFP compared to that of control.Conclusions Adiponectin antagonizes stimulatory effect of platelet-derived growth factor on mesangial cell proliferation by AMPK signaling.
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<p><b>OBJECTIVE</b>To introduce a new technique for rapid construction of gene site-directed mutagenesis.</p><p><b>METHODS</b>Three primers are synthesized. One is a primer with the needed mutation; the other two containing appropriate enzyme sites for construction of the PCR fragment into a suitable plasmid are located at the flanks of the mutation primer. After the amplification of the PCR fragment using the mutation primer and the reverse flanking primer, another PCR is performed using the previous PCR mutation segment as primer and the other flanking primer. The final PCR segment can be cloned into an appropriate plasmid by using the enzyme sites in the primers.</p><p><b>RESULTS</b>Two site-directed mutagenesis have been successfully constructed in the Parkin gene by this method.</p><p><b>CONCLUSION</b>The method is effective and simple for construction of gene site-directed mutagenesis.</p>