AIM:Vascular smooth muscle cell (SMC) proliferation and migration from the arterial wall media into the intima are believed to play a critical role in the pathogenesis of restenosis. Several studies have demonstrated that phosphothioate (PS) oligodeoxynucleotides targeted against genes involved in SMC proliferation inhibits in vitro SMC proliferation and migration. However, the therapeutic effect of antisense ODN on the individual who receives the treatment of delivery of the agent depends on the efficacy of this agent in great degree. We investigated the inhibition effect of a novel agent, insulin-antisense-c-myb-PS-ODN on SMC proliferation in vitro.　METHODS：The rat aortic artery SMCs were cultured in Dulbecco's modified Eagel's medium. The passage 8 to 13 were used as the experiment. Cell surface receptor binding assay was quantified through counting gamma particles emitted from 125　　　　I labeled insulin. SMC rapid proliferation was brought by stimulation of high concentration of fetal bovine serum (FBS). The novel agent of insulin conjugated to the antisense-c-myb-PS-ODN was obtained via incubation of both in condition of certain reagents, pH, temperature, and ion concentration. The characterization and purification of the agent was performed through HPLC. Inhibition of SMC proliferation was reflected by incorporation rate of trillium labeled thymidine deoxyribonucleotide.RESULTS：The binding efficacy of insulin to the receptor was remarkably increased in SMC cultured in supplement of 20% FBS. The inhibition effect of conjugator insulin-c-myb-antisense-PS-ODN was stronger than that of the simple c-myb-antisense-PS-ODN. The inhibition rate of conjugator and simple form on SMC proliferation were 48.34% and 29.54%, respectively. CONCLUSION：The binding efficacy and specificity of c-myb-antisense-PS-ODN to SMC may be enhanced by the insulin receptor mediation through the insulin-insulin receptor interaction. The insulin-receptor targeted method may be a potential and specific therapeutic pathway for restenosis.

Objective By analyzing and commentating the current research situation of recycling of residues in China, to provide reference for correlational research in future. Methods By taking periodical literature and national patent literature named residues at home and abroad as sample, using content analysis approach, the thesis conducts researches and analysis by computer statistical function. Results The literature mainly distributed in such branches as organic chemical industry, gardening, animal husbandry and animal medicine. The research involves residues and feedstuff, edible mushrooms, fertilizer, and analysis and application of the ingredients of residues. The types of residues mainly include Radix Liquorice, Radix Salviae Miltiorrhizae and brag-zhun, etc. And more than ten kinds of technologies of processing and curing residues were involved. Conclusion Research of recycling of residues in China is insufficient, the difficulty lies in the recycling of antibiotic residues. There will be a bright prospect in resource utilization of residues.

Objective To observe the clinical effect of Salmeterol xinafoate and fluticasone propionate powder for inhalation in patients with chronic obstructive pulmonary disease.Methods 70 patients diagnosed with chronic obstructive pulmonary disease from March 2014 to March 2015, were randomly divided into two groups ( n =35 ) .Control group were given basic treatment, observation group was given Salmeterol xinafoate and fluticasone propionate powder for inhalation on the basis of control group , patients were followed up and changes of related indicators wererecorded. Results After treatment one month, serum airway remodeling index b-FGF, TIMP-1 values were (93.86 ±17.36 μg/L, 38.06 ±4.28ng/mL) respectively, more than control group(135.03 ±16.06μg/L, 53.95 ±4.15ng/mL)(P<0.05).After treatment one month, inflammatory markers IL-8, TNF-αwere(7.26 ±1.57 pg/mL, 4.29 ±1.02 ng/L)respectively, were more than the control group (14.27 ±1.71 pg/mL, 8.90 ±1.21 ng/L) (P<0.05).After treatment, the observation group, the total effective rate was 91.43%, higher than 77.14%(P <0.05).Conclusion Salmeterol xinafoate and fluticasone propionate powder for inhalation has good effect in treatment of chronic obstructive pulmonary disease , better than the use of basic treatment alone.

Objective To investigate the association of lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms with metabolic syndrome. Methods PCR-RFLP was used to detect lipoprotein lipase Hind Ⅲ and S447X genotypes in 401 subjects(including 201 controls, 200 metabolic syndrome patients). Results ( 1 ) The levels of waist circumference ( WC ) , hip circumference ( HC ) , waist-to-hip ratio ( WHR ) , body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure ( DBP) , total cholesterol ( TC) , triglyceride (TG), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and fasting plasma glucose (FPG) were significantly different between metabolic syndrome group and control group (all P< 0.05). (2)The frequencies of H+H+ genotype,H+allele,SS genotype, and S allele for metabolic syndrome were all significantly higher than those for controls( H+H+ genotype:66. 5% vs 54.2% ,P=0.012; H+ allele:78.0% vs 71.4%, P=0.031;SS genotype:89.5% vs 77. 1% , P = 0.001; Sallel:94.5% vs 87. 56% , P = 0.001). (3) The levels of WC, HC, WHR, BMI, SBP, DBP, TG, LDL-C, and FPG in H + H-/H-H- genotype were significantly lower than those in H+H+ genotype, HDL-C was significantly higher than that in H+H+ genotype ( all P<0. 05). The levels of WC, HC, WHR, BMI, SBP, DBP, TC, TG, and FPG in SX/XX genotype were significantly lower than those in SS genotype, HDL-C was significantly higher than that in SS genotype ( all P< 0.05). (4)Multi-way logistic regression analysis suggested that risk factors for metabolic syndrome were smoking, drinking, and SS genotype (OR value was 4.289,2.268, and 2. 597, respectively ). (5) Result of interaction analysis among different factors indicated that the risk for metabolic syndrome in smoker with SS genotype was 3. 996 times of non-smokers with SX/XX genotype. Conclusions The lipoprotein lipase gene S447X polymorphism is associated with metabolic syndrome risk in Kazakh, and SS genotype and S allele may serve as genetic risk factors of metabolic syndrome, H + H-/H-H- and SX/XX genotypes yield beneficial effect for lipid and blood pressure. SS genotype and smoking may exist additive effect.

ObjectiveTo evaluate the efficacy and safety profiles of enteeavir (ETV) in patients with advanced schistosomiasis and hepatitis B virus (HBV) co-infection.Methods Totally sixty patients with advanced schistosomiasis and HBV co-infection were enrolled in this study.The patients were divided into ETV treatment group (n=30) and rhubarb treatment group who refused to receive antiviral treatment (n=30).The patients were treated with ETV or rhubarb thelepus ball on the basis of routine supportive therapy for 52 weeks.The hepatic fibrosis markers (e.g.hyaluronic acid,type Ⅲ procollagen,type Ⅳ collagen,laminin and fibronectin),alanine transaminase (ALT),HBV DNA,Child-Pugh score between two groups were compared.Intention to treat (ITT) population was used for analysis.The measurement data and the enumeration data were analyzed by t test and x2 test,respectively.ResultsAfter 52-week treatment,the hepatic fibrosis markers (hyaluronic acid,type Ⅲ procollagen,type Ⅳ collagen,laminin and fibronectin) were significantly improved in ETV treatment group compared to the rhubarb treatment group (t =3.952,3.765,3.857,3.122 and 3.735,respectively; all P＜0.05),and the fibrosis of liver tissue in ETV treatment group was significantly improved compared with rhubarb treatment group (x2 =11.207,P＜0.05).The ALT level,HBV DNA,Child-Pugh score after 52-weeks treatment in ETV treatment group were statistically reduced compared with rhubarb treatment group (t =3.287,4.382 and 3.872,respectively; all P＜0.05),meanwhile,the ALT normalization rate and HBV DNA undetectable rate were significantly increased in ETV treatment group (x2 =17.376 and 39.095,respectively; both P＜0.05).In addition,no obvious adverse reaction was observed during ETV treatment.Conclusion Entecavir is safe and effective in patients with advanced schistosomiasis and HBV co-infection.

OBJECTIVETo identify the genetic etiology in a Chinese patient with neurofibromatosis type 1 (NF-1).

METHODSAll coding exons and the flanking sequences of neurofibromin 1 (NF1) gene from the patient were captured, individually barcoded and subjected to HiSeq2000 high-throughput sequencing. Suspected mutation was validated in the nuclear family members with Sanger sequencing.

RESULTSA novel indel mutation, c.789_790delAGinsT, was identified in the exon 8 of the NF1 gene in the patient but not in her asymptomatic parents. The mutation was predicted to have caused shifting of the reading frame and a premature downstream stop codon (p.K263Nfs*18). Two known polymorphisms, c.888+108 C>T (rs2953000) and c.888+118 G>T (rs2952999), was detected in the flanking of the indel mutation in the patient and her father. Sequencing chromatogram for the family indicates that above changes are located on the same chromosome.

CONCLUSIONThe c.789_790delAGinsT, as a de novo mutation occurring on the paternally derived chromosome, is most likely to be causative for the disease. Compared with Sanger sequencing, targeted next-generation sequencing is more efficient and can dramatically reduce the cost for the genetic testing of NF-1.

Adult ; Amino Acid Sequence ; Base Sequence ; Female ; Humans ; Molecular Sequence Data ; Neurofibromatosis 1 ; enzymology ; genetics ; Neurofibromin 1 ; genetics ; metabolism ; Point Mutation

Objective:To compare the clinical effects of percutaneous curved vertebroplasty (PCVP) and unilateral percutaneous kyphoplasty (PKP) in the treatment of osteoporotic vertebral compression fracture (OVCF).Methods:A retrospective cohort study was used to analyze the clinical data of 104 patients with single vertebral OVCF treated in Tianjin Hospital from September 2019 to September 2020, including 21 males and 83 females; aged 50-91 years [(70.3±7.7)years]. AO classification of the fracture was type A1 in 65 patients and type A2 in 39. The patients received PCVP (PCVP group, n=51) or unilateral PKP surgery (unilateral PKP group, n=53). The operation time, bone cement injection volume, intraoperative fluoroscopy frequency, effective dispersion times of bone cement and excellent rate of bone cement distribution were compared between the two groups. In evaluation of the therapeutic effects of the two groups, visual analogue scale (VAS) and Oswestry dysfunction index (ODI) were measured preoperatively and at postoperative 24 hours, 3 months and 6 months; Beck index was measured preoperatively and at postoperative 24 hours and 3 months. The rate of bone cement leakage and rate of refracture of adjacent vertebral bodies were compared between the two groups. Results:All patients were followed up for 6-8 months [(6.4±0.7)months]. The operation time, bone cement injection volume and intraoperative fluoroscopy frequency in PCVP group was (12.15±1.63)minutes, (2.13±0.28)ml and (24.74±1.71)times, shorter or less than (22.09±1.62)minutes, (5.30±0.52)ml and (30.09±1.86)times in unilateral PKP group (all P<0.01). The effective dispersion times of bone cement in PCVP group was (1.42±0.04)times, higher than (1.18±0.02)times in unilateral PKP group ( P<0.01). The excellent rate of bone cement distribution in PCVP group was 94%, higher than 70% in unilateral PKP group ( P<0.01). There were no significant differences in VAS, ODI and Beck index between the two groups before operation and at 24 hours and 3 months after operation (all P>0.05). VAS and ODI in PCVP group were (1.20±0.49)points and 16.52±5.22 at 6 months after operation, lower than (1.49±0.58)points and 20.16±5.16 in unilateral PKP group (all P<0.01). VAS and ODI in the two groups were significantly improved at 24 hours, 3 months and 6 months after operation when compared with those before operation (all P<0.05). Beck index in the two groups detected at 24 hours and 3 months after operation was improved from that before operation (all P<0.05). Unilateral PKP group showed Beck index was 0.75±0.07 at 3 months after operation, significantly lower than 0.79±0.07 at 24 hours after operation ( P<0.05), but there was no significant change in PCVP group ( P>0.05). The leakage rate of bone cement in PCVP group was 16% (8/51), lower than 47% (25/53) in unilateral PKP group ( P<0.01). There was no significant difference in the incidence of refracture of adjacent vertebral bodies between the two groups during follow-up ( P>0.05). Conclusion:For OVCF, PCVP is superior to unilateral PKP in terms of operation time, amount of bone cement injection, intraoperative fluoroscopy frequency, dispersion effect of bone cement in vertebral body, pain, function improvement, maintenance of injured vertebral height and incidence of bone cement leakage.

OBJECTIVETo detect the most prevalent mutations, R778L and P992L of ATP8B gene, in Chinese Wilson disease(WD) patients by high resolution melting (HRM) analysis after polymerase chain reaction (PCR).

METHODSGenomic DNA was extracted from peripheral blood samples obtained from 30 cases of WD by the standard phenol/chloroform method. DNA fragments encompassing ATP7B exons 8 and 13 were produced by PCR amplification. The amplicons containing the R778L or P992L mutations were then generated by nested PCR. The nested PCR products were subjected to HRM analysis using the HR-1 instrument. Mutations detected in HRM analysis were verified by restriction analysis using restriction enzyme (MspI or AluI or AfaI) or DNA sequencing.

RESULTSHRM analysis of the fragments encompassing ATP7B exon 8 showed four curve patterns. Subsequent restriction analysis and DNA sequencing proved that the four different curves represent four different genotypes: the wild type, the R778L/R778L homozygote, the R778L heterozygote, and the R778L/752.33delG compound heterozygote. Three HRM curve patterns were observed for the fragments encompassing ATP7B exon 13, representing the wild type, the P992L heterozygote, and the P992L/S975Y compound heterozygote. In our studied samples, allele frequencies of the R778L, P992L and S975Y mutations were 25%, 15% and 1.67%, respectively.

CONCLUSIONHRM analysis is a simple, accurate and sensitive approach for rapid detection of the ATP7B mutations and could be used as an optimized method for genetic testing in WD.

Adenosine Triphosphatases ; genetics ; Base Sequence ; Cation Transport Proteins ; genetics ; Copper-transporting ATPases ; DNA ; genetics ; metabolism ; DNA Mutational Analysis ; methods ; DNA Restriction Enzymes ; metabolism ; Exons ; genetics ; Freezing ; Gene Frequency ; Genotype ; Hepatolenticular Degeneration ; genetics ; Humans ; Nucleic Acid Denaturation ; Polymerase Chain Reaction ; Time Factors

Objective:To investigate the therapeutic effect of bidirectional-traction, Steinmann pin poking reduction,anatomic plate and raft technique in the treatment of hyperextension tibial plateau fractures.Methods:The data of 25 patients with hyperextension tibial plateau fractures admitted to Qilu Hospital of Shandong University (Qingdao) from July 2017 to June 2022 were retrospectively analyzed. According to the treatment methods, they were divided into bidirectional-traction group (treated with bidirectional-traction, Steinmann pin poking reduction, anatomic plate and raft technique) and open reduction group (treated with open reduction, bone grafting and two plates fixation). The bidirectional-traction group included 14 patients, with 8 males and 6 females; the age was 50.29±9.23 years (range, 38-61 years). The cause of the injury was a traffic accident in 4 patients, a fall from height in 7 patients and a fall from standing height in 3 patients. According to Schatzker classification, there were 5 Schatzker type V and 9 type VI fractures. The open reduction group included 11 patients (7 males and 4 females); with a mean age of 58.00±10.58 years (range, 48-69 years). 3 cases were injured by traffic accident, 6 cases by falling from height, and 2 cases by falling from standing height. According to Schatzker classification, there were 4 type V and 7 type VI. Preoperative waiting time, operative time, blood loss and percentage of blood loss, incision length, fracture healing time, tibial posterior inclination, medial proximal tibial angle, visual analogue scale (VAS) on the first day after surgery, Hospital for Special Surgery (HSS) score 6 months after surgery were compared between the two groups.Results:Patients in both groups were followed up for more than 6 months. The follow-up time was 7 to 48 months with an average of 22.76 months. There were significant differences in the preoperative waiting time [6 (4, 8) d vs. 8 (7, 11) d, W=114.00, P=0.043], the incision length [15.0 (12.5, 16.0) cm vs. 30.0 (28.0, 31.0) cm, W=154.00, P<0.001], postoperative VAS [4 (3, 4) points vs. 5 (5, 6) points, W=143.00, P<0.001], blood loss [147 (107, 206) ml vs. 267 (191, 362) ml, W=116.00, P=0.033], blood loss percentage [2.95% (2.58%, 5.20%) vs. 6.40% (4.05%, 7.00%), W=118.00, P=0.027] between the bidirectional-traction group and open reduction group. There were not significant differences in the operation time [120 (118, 120) min vs. 119 (101, 154) min, W=68.50, P=0.656], fracture healing time (8.18±1.03 weeks vs. 8.86±1.27 weeks, t=1.49, P=0.149), HSS score (8.43±3.72 vs. 85.18±7.73, t=1.28, P=0.221) and medial proximal tibial angle 6 months after surgery (87.66°±1.53° vs. 86.47°±2.24°, t=1.57, P=0.130) between the two groups. Postoperative tibial posterior inclination was improved in both groups. There was no significant difference in the tibial posterior inclination before surgery, immediately after surgery and 6 months after surgery (-14.96°±6.44°, 5.55°±1.02°, 5.61°±0.82°) in the bidirectional-traction group and -12.26°±2.93°, 7.07°±3.21° and 7.14°±3.17° in the open reduction group, P>0.05). There were no postoperative complications such as acute compartment syndrome or knee stiffness in both groups. Conclusion:The treatment of hyperextension tibial plateau fracture with bidirectional-traction, Steinmann pin poking reduction, anatomic plate and raft technique can shorten preoperative waiting time, reduce incision length, decrease blood loss and lower VAS. It is a minimally invasive, rapid and effective method, which has achieved good clinical results and is worth promoting.

OBJECTIVETo characterize the mutation spectrum of phenylalanine hydroxylase (PAH) gene and perform prenatal diagnosis for families with classical phenylketonuria.

METHODSBy stratified sequencing, mutations were detected in the exons and flaking introns of PAH gene of 44 families with classical phenylketonuria. 47 fetuses were diagnosed by combined sequencing with linkage analysis of three common short tandem repeats (STR) (PAH-STR, PAH-26 and PAH-32) in the PAH gene.

RESULTSThirty-one types of mutations were identified. A total of 84 mutations were identified in 88 alleles (95.45%), in which the most common mutation have been R243Q (21.59%), EX6-96A>G (6.82%), IVS4-1G>A (5.86%) and IVS7+2T>A (5.86%). Most mutations were found in exons 3, 5, 6, 7, 11 and 12. The polymorphism information content (PIC) of these three STR markers was 0.71 (PAH-STR), 0.48 (PAH-26) and 0.40 (PAH-32), respectively. Prenatal diagnosis was performed successfully with the combined method in 47 fetuses of 44 classical phenylketonuria families. Among them, 11 (23.4%) were diagnosed as affected, 24 (51.1%) as carriers, and 12 (25.5%) as unaffected.

CONCLUSIONPrenatal diagnosis can be achieved efficiently and accurately by stratified sequencing of PAH gene and linkage analysis of STR for classical phenylketonuria families.

Adolescent ; Adult ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Fetal Diseases ; diagnosis ; enzymology ; genetics ; Genetic Testing ; Humans ; Infant ; Infant, Newborn ; Male ; Microsatellite Repeats ; Middle Aged ; Pedigree ; Phenylalanine Hydroxylase ; genetics ; Phenylketonurias ; diagnosis ; enzymology ; genetics ; Point Mutation ; Pregnancy ; Prenatal Diagnosis ; Young Adult