Objective To clone, express and purify human PIF1 protein and analyze its ATPase activity. Methods The PIF1 cDNA was amplified by PCR from HeLa cell cDNA library and inserted to pET24b with histidine tag at its terminus to form pET24b-PIF1 plasmid. The recombinant pET24b-PIF1 plasmid was transformed to RosettaTM 2 (DE3) and the expression of PIF1 protein was monitored by SDS-PAGE analysis. By using fast protein liquid chromatograph (FPLC) system, the PIF1 protein was purified by affinity chromatograph and gel filtration. The ATPase activity of PIF1 was checked by thin layer chromatograph(TLC). Results The PIF1 protein was successfully cloned and expressed in E.coli. Conclusions The purification procedure of PIF1 protein was established using FPLC. The overexpressed and the purified PIF1 helicase has DNA and Mg2+ dependent ATPase activity.

Heart Septal Defects, Atrial ; complications ; Humans ; Infant, Newborn ; Male ; Nasal Polyps ; complications ; congenital ; Nasopharyngeal Diseases ; complications ; congenital

Objective:To study the association between rs2235371,rs2013162,rs2235377 SNPs in interferon regulatory factor 6 (IRF6)gene and non-syndromic cleft lip with or without cleft palate(NSCL/P)in Xinjiang Uyghur population.Methods:100 Uyghur NSCL/P patients from Xinjiang were included in the case group and 60 Uyghur inpatients with upper respiratory tract infection were se-lected in the control group.Next,generation sequencing was used,DNA sequencing results were compared with the information on the genome database and genetic analysis were made.Results:There were no significant differences in the frequency distribution of both genotypes and alles when the cases were campared with the controls at the rs2235371,rs2013162 and rs2235377 loci(P >0.05). Above three loci were located in the same block,rs2235371 and rs2235377 loci presents the strong linkage disequilibrium(r2 =0.949, D'=0.974).Possible haplotypes were:CCT >CAT >TAC,and there was no significant difference between the cases and controls in haplotype distribution(P >0.05).Conclusion:Polymorphisms of rs2235371,rs2013162 and rs2235377 in IRF6 gene may be associ-ated with NSCL/P in Xinjiang Uygur people.

Objective To investigate the changes and the correlation between the levels of plasma lysophosphalidic acid(LPA),acid phospholipid(AP),low density lipoprotein(LDL) and carotid artery atherosclerosis in patients with acute cerebral infarction(ACI).Methods The levels of plasma LPA,AP and LDL were examined in 67 ACI patients and 30 normal controls,the carotid arteries were investigated with B mode ultrosand.Results The levels of plasma LPA,AP and LDL were higher in ACI group than those in the normal control group(all P

Objective:The aim of this study was to investigate the carrier ratio and the genotype of thalassemia among Tujia and Miao people of reproductive age in Chongqing.Methods:According to forward-looking design and multi-stage stratified cluster sampling method, fasting venous blood samples of Tujia and Miao people of reproductive age were collected from 11 survey sites in Chongqing from March to July 2019. Gap-PCR and high-throughput sequencing were used to screen thalassemia genes.Results:A total of 516 Tujia people (258 males, 258 females) and 270 Miao people (139 males, 131 females) were included in this study, and their age were (28.63 ± 5.26) and (28.62 ± 5.35) years, respectively. About 5.04% (26/516) Tujia people carried thalassemia gene, with 1.94% (10/516) and 2.52% (13/516) for α and β thalassemia, respectively. Three kinds of new variants (1 case of each variant), HBA 2: c.46G>A (Gly>Ser), HBB: c.*+129T>A and HBB: c.-39T>G with unclear pathogenicity, were identified in Tujia people. About 7.78% (21/270) Miao people carried thalassemia gene, among these, α and β thalassemia were 3.33% (9/270) and 4.44%(12/270), respectively. The most common mutation type of α-globin gene was -α 3.7/in the two ethnic groups. Three kinds of β-globin gene mutation types, Codons 41/42 (-TTCT) beta 0, Codon 17 (A>T) beta 0 and IVS-Ⅱ-654 (C>T) beta +, were the most common in Tujia people. Meanwhile, the chief β-globin gene mutation type was Codons 41/42 (-TTCT) beta 0 in Miao people. Conclusions:The carrying rate of thalassemia gene is higher in Tujia and Miao people in Chongqing, and the genotypes of thalassemia gene are different between Tujia and Miao people. The clinical significance of three kinds of new variants with unclear pathogenicity should be focused on.

The clinical data of a case of Weave syndrome admitted in the Department of Endocrinology, Children′s Hospital of Nanjing Medical University in October 2018 were retrospectively analyzed.The patient was a 9 years and 2 months old girl, who was hospitalized because of " growing too fast for 9 years" . After birth, the child is found to grow fast and have mental retardation, slurred speech, a blurred vision, a long face, a protruding forehead, ocular hypertelorism, epicanthus, nasal bridge pit, finger pads on both hands and feet, uncoordinated gaits, and intoeingpigeon toes.A novel heterozygous c. 1720A>G (p.K574E) mutation was detected in the exon 15 of the EZH2 gene of the patient.This mutation has not been reported at home and overseas.Sanger sequencing revealed that the patient′s parents did not carry the mutation.The disease is an autosomal dominant genetic disorder, and the parents and sibling of the patient have no corresponding symptoms, so it is inferred that the mutation is spontaneous.Based on the peculiarity of the face, clinical manifestations and the results of molecular genetics, the child was diagnosed as Weaver syndrome.

OBJECTIVE: To study the quantitative determination method of ferulic acid in the Wumeitougu oral liquid,compare the stability of ferulic acid in different solvents. METHODS:The detecting conditions of RP -- HPLC were as followKromasil C_(18) column(4.6mm?200mm, 5?m); mobile phase: methanol-- water(30:70); flow rate: 1ml/ min: detecting waslength: 324nm. RESULTS: The calibration curve revealed linearity in the range of 0.05～1.0?g(r=0.9 999). The mean re?ery was 97.01%～101.53%. RSD

Purpose:To study the relation between regional lymph node metastasis and nm23 H 1 protein expression and DNA ploidy of esophageal squamous cell carcinoma patients. Methods:Flow cytometry was used to analyze nm23 H 1 protein and DNA ploidy in surgical specimens from the patients with esophageal carcinoma. The object tissues included: the primary tumor tissue, the pericancerous mucosa, the incisional margin and the regional lymph node. Results:There were obvious differences between nm23 H 1 protein expression in lymph node metastasis positive and negative groups, primary tumor tissue and pericancerous mucosa, cancer tissue and the incisal margin( P 0.05).The differences of DI were distinct between cancer tissue and the pericancerous mucosa, cancer tissue and the incisal margin, cancer tissue and the lymph node( P 0.05).In corresponding tissues of lymph node metastasis positive and negative groups, nm23 H 1 protein expression was different( P 0.05). As to DI, it was different only among pathological grades( P 0.05). Conclusions:nm23 H 1 protein higher expression can produce certain inhibitory effect on metastasis of esophageal carcinoma. Combining DNA ploidy detection can help to understand the biological behavious and deduce the prognosis of esophageal carcinoma.

Objective To explore the health-relate quality of life(QOL) status of adult persistent allergic rhinitis (PAR);the change of QOL of pro-post specific immunotherapy (SIT) and pharmacotherapy .Methods Skin prick tests(SPT) were performed on PAR patients .According to the results ,80 adult cases that were allergic to dermatophagoides were enrolled in ENT outpatient clinic of affiliated hospital of Nantong University from April to August 2011 .The patients were randomly allocated to receive either specific immunotherapy(n=40) or pharmacotherapy (n=40) ,all of them were given RQLQ before and after half-year treatment ;40 cases without any allergic diseases were chosen from ENT in-patient department ,and were given RQLQ .The scores of previous treatment of the PAR group were compared with health control group ,then compared with the scores of post-treatment ,and also compared the scores of post-treatment between the immunotherapy group and pharmacotherapy group .Results The scores of the PAR patients were higher than that of health control patients in all dimensions of RQLQ (P< 0 .05) ,and the most troublesome problems were nasal symptoms .The scores of the patients who received SIT were evidently lower than that of pro-treatment in all dimensions of RQLQ(P<0 .05) ,the scores of the patients who received medical treatment were also lower than before (P<0 .05) , and the scores of the SIT group were lower than the pharmacotherapy group (P<0 .05) .Conclusion The QOL of adult patients with PAR was improved after SIT or drug treatment ,and QOL improvement is more obvious treat by SIT .