Objective To investigate the effect of nicotine on coagulation in intestinal ischemia-reperfusion injury rats.Methods 32 male Sprague-dawley rats,weighing 250-300g,were randomly divided into 4 groups(n=8):sham operation group(S),intestinal ischemia-reperfusion(IR)group,nicotine(NIC)group,α7 nicotinic acetylcholine receptor(α7nAchR)antagonist group α-bungarotoxin(α-BGT)group.Intestinal IR was induced by clamping superior mesenteric artery for 45min and 120min of reperfusion.In group NIC nicotine 400μg/kg was injected intraperitoneally at 30min before superior mesenteric artery occlusion.In group α-BGT 1μg/kg was injected intraperitoneally at 15min before superior mesenteric artery occlusion.Plasma tumor necrosis factor-α(TNF-α),tissue factor(TF),antithrombin(AT),tissue plasminogen activator(tPA),fiber plasminogen activator inhibitor-1(PAI-1),D-dimer levels and platelet count(PLT)were measured after 120min reperfusion.Chiu's count was used to assess the changes in intestinal mucosal pathlolgical morphology.Results Compared with group S and group NIC,the plasma TNF-α,TF,tPA,PAI-1 and D-dimer levels were significantly increased,and plasma AT level and platelet count were significantly decreased,in group IR and group α-BGT(P<0.05),Chiu's scores were significantly increased(P<0.05).Conclusion Nicotine can inhibit the excessive activation of coagulation function in intestinal ischemia-reperfusion injury rats.Its mechanism may be related to activation of cholinergic antiinflammatory pathway,reducing the release of pro-inflammatory cytokines thereby reducing endothelial cell injury.

Whether direct manipulation of Parkinson's disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6-10 months), and thus provides a practical transgenic monkey model for future PD studies.
Animals ; Brain ; CRISPR-Cas Systems/genetics* ; Dependovirus/genetics* ; Haplorhini ; Phenotype ; Protein Kinases/genetics*

Whether direct manipulation of Parkinson’s disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6–10 months), and thus provides a practical transgenic monkey model for future PD studies.

Congenital heart disease (CHD) is the most common congenital malformation disease in infants.With the rapid development of clinical treatment, the survival rate of children with CHD increased significantly.However, postoperative complications in children with CHD, including heart failure, heart arrythmia, pulmonary hypertension and brain cognitive dysfunction, are another challenge, among which, brain cognitive dysfunction has been gradually recognized and valued by scholars.At present, the effect of cardiopulmonary bypass on brain cognitive function of children with CHD has been studied by a variety of research methods, including molecular biological technique, magnetic resonance imaging technique, optical imaging technology, brain electrical activity monitoring technology and nervous development scale.Varying degrees of brain cognitive dysfunction will occur in children with CHD after heart surgery, which is closely associated with many factors such as systemic inflammatory response related to cardiopulmonary bypass, cerebral ischemia-reperfustion injury, the duration of cardiopulmonary bypass, aortic cross clamp time, deep hypothermia circulatory arrest time, dynamic changes of temperature, degree of hemodilution and blood gas management strategy.The influence of cardiopulmonary bypass on postoperative brain cognitive function in children with CHD is reviewed in this paper, in order to provide reference for clinical diagnosis and treatment, and to improve the prognosis of children with CHD.

Objective To analyze the variation characteristics of rpoB,katG,inhA,rpsL and embB related genes of Mycobacterium tuberculosis(MTB)in Qinzhou,Guangxi.Methods PCR reverse point hybridization was used to detect 5 common resistance mutants of Mycobacterium tuberculosis in 237 MTB-DNA positive sputum samples.Results Among 237 cases of tuberculosis patients,72 cases(30.38%)were resistant to the four kinds of anti-TB drugs.The resistance mutation rate of rifampin,isoniazid and streptomycin was 2.53%, 13.92%,3.80%.The top 5 gene mutation detection loci of Mycobacterium tuberculosis were-15M,S531L and 43M.Conclusion The main drug-resistant strains are isoniazid resistance,and the mutation of inhA gene were the major one in Qinzhou,Guangxi.

Objective To investigate the clinical effect of anterior controllable antedisplacement and fusion (ACAF) for the treatment of ossification of the posterior longitudinal ligament (OPLL) of the cervical spine.Methods The data of 45 cases with cervical posterior longitudinal ligament ossification treated by ACAF from March 2017 to October 2017 were retrospectively analyzed,including 25 males and 20 females,age 45-68 years,average 57.5 years.There were 18 cases involving C3 vertebral body,30 cases involving C4 vertebral body,40 cases involving C5 vertebral body,34 cases involving C6 vertebral body,and 7 cases involving C7 vertebral body.The function of the neural function was evaluated by the Japanese Orthopaedic Association (JOA) scoring system at preoperation and latest follow-up.The curvature of the cervical spine was measured on the lateral X-ray film of the cervical spine,the maximum occupying ratio of the spinal canal was measured on the cross section of the CT scan,and compression of the cervical spinal cord was evaluated by the cervical MRI.Results Patients were followed up for 3 to 6 months (average,3.9 months).The improvement of neurological function was obtained in all the patients.The JOA score improvement rate at the latest follow-up was 71.3％±9.6％.The cervical lordosis was improved from preoperative 4.5°±3.8° to 10.3°±4.8° at the latest follow-up.The canal stenosis ratio was decreased from preoperative 54.3％±8.2％ to 12.5％±5.3％ at the latest follow-up.MRI showed that the cervical spinal cord was adequately decompressed in situ.No specific complications were identified that were associated with this technique.Conclusion The present study elaborates the surgical tips and demonstrates the satisfactory outcome of ACAF for the treatment of OPLL.This novel technique has the potential to serve as an alternative surgical technique for the treatment of cervical OPLL.

OBJECTIVETo explore the value of single nucleotide polymorphism array (SNP-array) for the analysis of pediatric patients with growth retardation.

METHODSOne hundred eighty one children with growth retardation were enrolled. DNA was extracted from peripheral samples from the patients, and whole genome copy number variations (CNVs) were detected using Illumina Human Cyto SNP-12. All identified CNVs were further analyzed with reference to databases including ClinGen, ClinVar, DECIPHER, OMIM and DGV as well as comprehensive review of literature from PubMed to determine their pathogenicity.

RESULTSForty seven patients (26%) with abnormal CNVs were detected, which included 12 known microdeletions/microduplications syndrome (26%), 10 pathogenic non-syndromic CNVs (21%), 3 numerical chromosome aberrations (6%), 3 unbalanced translocations (6%), 4 pathogenic mosaicisms (9%) and 15 cases with unknown clinical significance (32%). After excluding obvious numerical and/or structural chromosomal abnormalities, this study has detected 15 pathogenic microdeletions/microduplications sized 5 Mb or less, which may be missed by routine chromosomal karyotyping. In addition, there were 3 cases with loss of heterozygoisty (LOH) containing known or predicted imprinting genes as well as 2 cases with suspected parental consanguinity.

CONCLUSIONSNP-array technology is a powerful tool for the genetic diagnosis of children with growth disorders with advantages of high resolution and improved accuracy.

Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; DNA Copy Number Variations ; Developmental Disabilities ; diagnosis ; genetics ; Female ; Humans ; Infant ; Karyotyping ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Polymorphism, Single Nucleotide

Objective To evaluate the effect of electro-acupuncture (EA) at Zusanli (ST36) acupoint on blood coagulation during intestinal ischemia-reperfusion (I/R) in rats.Methods Forty healthy male Sprague-Dawley rats,aged 6-8 months,weighing 250-300 g,were divided into 5 groups (n =8 each) using a random number table:sham operation group (group S),intestinal I/R group (group I/R),EA at Zusanli acupoint group (group EA),EA at non-acupoint group (group NE) and α7 nicotinic acetylcholine receptor antagonist α-bungarotoxin (α-BGT) group (group α-BGT).Intestinal I/R was induced by clamping the superior mesenteric artery for 4-5 min followed by 120 min of reperfusion.Bilateral Zusanli acupoints were stimulated with an electric stimulator (frequency 3 Hz,voltage 2-4 V,wave length 2 ms) for 30 min starting from the time point immediately after beginning of ischemia in group EA,while EA was performed at the points 5 mm lateral to the bilateral Zusanli instead in group NE.In group α-BGT,α-BGT 1 μg/kg was intraperitoneally injected at 45 min before ischemia,and the other treatments were similar to those previously described in group EA.Blood samples were collected from the abdominal aorta at 120 min of reperfusion for determination of the concentrations of tumor necrosis factor alpha (TNFα),tissue factor (TF),antithrombin (AT),tissue plasminogen activator (tPA),fiber plasminogen activator inhibitor-1 (PAl-l) and D-dimer in plasma (by enzyme-linked immunosorbent assay) and platelet count (PLT).The animals were sacrificed after blood sampling,the distal ileum specimens were removed for examination of the pathological changes with a light microscope,and the damage to the intestinal mucous membrane was assessed and scored according to Chin.Results Compared with group S,the concentrations of plasma TNFα,TF,tPA,PAI-1 and D-dimer were significantly increased,and the plasma AT concentration and PLT were decreased in I/R,NE and α-BGT groups,the concentrations of plasma TNFα and TF were significantly increased,and the plasma AT concentration was decreased in group EA,and Chiu's scores were significantly increased in I/R,EA,NE and α-BGT groups (P＜ 0.05).Compared with group I/R,the concentrations of plasma TNFα,TF,tPA,PAI-1 and D-dimer were significantly decreased,the plasma AT concentration and PLT were increased,and Chiu's scores were decreased in group EA (P＜0.05),and no significant change was found in the variables mentioned above in NE and α-BGT groups (P＞0.05).Compared with group EA,the concentrations of plasma TNFα,TF,tPA,PAI-1 and D-dimer were significantly increased,the plasma AT concentration and PLT were decreased,and Chiu's scores were increased in group NE (P＜0.05).Conclusion EA at Zusanli acupoint can improve blood coagulation during intestinal I/R in rats,and the mechanism is related to activating the cholinergic anti-inflammatory pathway.

Objective:To determine the prevalence, disease burden, and possible causes of lung cancer. Population-based lung cancer registration data collected from 1993 to 2012 were used to analyze the lung cancer epidemic status in Qidong City. Methods:Joinpoint regression was used to calculate the trend of incidence and mortality for lung cancer. Age-period-cohort model was used to assess the effects of age and exposure on the incidence and mortality rates. Results:Upward trends were observed both for the incidence and mortality rates of lung cancer. The risk of incidence and mortality increased with age. According to the cohort effect, the risk of lung cancer increased with recent birth dates. Women had higher risk than men. Conclusion:The incidence and mortality rates of pulmo-nary neoplasms have annually increased, and the exposure to risk factors for lung cancer gradually increases. Among the community population in Qidong City, people over 70 years belong to the high-risk groups.

Objective To establish a study cohort of chronic hepatitis B (CHB) in Qidong and evaluate its baseline characteristics. Methods CHB outpatients of the Third People's Hospital of Qidong were invited to participate in baseline survey from January 1, 2016, including questionnaire survey, liver function detection, serum detection of HBV infection and upper abdomen ultrasound detection. Anticipated sample size was at least one thousand. Baseline data were inputted by EpiData 3.1 software and then cleaned and analyzed by SAS 9.3 software. Results As of 18 July, 2016, a total of 1006 participants had been enrolled into the current study, including 615 males with an average age of (44.26±9.97) years and 391 females with an average age of (46.66±11.17) years. The difference in family history of liver disease was not significant between males and females (P>0.05), while the differences in other key information, such as age, education level, tobacco consumption, alcohol drinking, tea consumption, and antiviral intervention, were significant between males and females (P<0.05). Among the key clinical parameters, such as ALT, HBeAg, HBsAg, HBV DNA, albumin, and width of splenic vein and portal vein, only the abnormal rates of ALT and total bilirubin levels were higher in males than in females, the difference was significant (P<0.05). Conclusion Outpatient department-based CHB cohort was established successfully in Qidong, and sub-cohort could be divided according to the differences on baseline characteristics.