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.

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

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

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.

Humans ; Medicine, Chinese Traditional ; methods ; Phytotherapy ; methods ; Spinal Cord Injuries ; therapy

Objective To determine the median effective concentration (EC50) of ropivacaine for ultrasound-guided brachial plexus block.Methods Fifty ASA Ⅰ or Ⅱ patients of both sexes, aged 19-72 yr, weighing 45-83 kg, scheduled for upper extremity surgery under brachial plexus block guided by ultrasound, were enrolled in this study. Brachial plexus block was performed under the guidance of ultrasound. After successful location, ropivacaine 30 ml was injected. EC50 of ropivacaine was determined by up-and-down sequential method. The initial concentration was 0.50% . Each time the concentration increased/decreased by 0.05% . EC50 of ropivacaine required for ultrasound-guided brachial plexus block and 95% confidence interval were calculated using Probit analysis.Results The EC50 of ropivacaine resulting in complete block of the brachial plexus nerve was 0.436%(95% confidence interval 0.393%-0.477% ). Conclusion The EC50 of ropivacaine is 0.436% for ultrasoundguided brachial plexus block.

Objective To investigate the pathogenic types and clinical features of children with lobar pneumonia. Methods Eighty children with lobar pneumonia diagnosed from April 2013 to May 2015 were enrolled. Bronchoalveolar lavage lfuid (BALF) of patients were collected. FQ-PCR was used to detect and analyze pathogens in BALF. Results In 80 cases, 59 cases were Mycoplasma pneumoniae, and 2 cases were Chlamydia pneumoniae, 12 cases were Streptococcus pneumoniae, 1 case was Klebsiella pneumoniae, 8 cases were adenovirus, 1 case was respiratory syncytial virus, 14 cases were of mixed infection. The prevalence of MP lobar pneumonia in children of 7-14 years old age group were higher than that of other age groups. Conclusions The pathogen of children with lobar pneumonia varied from Mycoplasma pneumoniae, Chlamydia pneumoniae, bacteria, virus and so on, and Mycoplasma pneumoniae was the common pathogen in lobar pneumonia.

Objective To retrospectively analyze computerized tomography (CT) imaging features of colorectal cancer with chronic schistosomiasis in order to improve the diagnostic accuracy.Methods Eighty patients whose diagnosis was pathologically confirmed as colorectal cancer with schistosomiasis were collected from Tongji Hospital and Kunshan Chinese Medicine Hospital from January 2007 to December 2012.All the patients underwent abdominal plain CT and contrast-enhanced CT scan.The lesion location,morphology,size,calcification features,enhancement patterns and cancer metastasis were evaluated and compared by two radiologists who were blind to the diagnosis.Twenty colorectal cancer cases without schistosomiasis from the same area were also collected as controls.Results CT imaging showed that the tumors all occurred in the colon and rectum in 80 patients,mainly in colon descendens,colon sigmoideum and rectum.The lesion was characterized by irregular bowel wall thickening with soft tissue masses,and the average length of impaired intestine was (21.35 ± 4.50) cm.The lesions were solitary in 68 cases (85.0％) and were multifocal in 12 cases (15.0％).Linear,spotty and small patchy calcifications were seen in all the patients,with margins unclear in 59 patients (73.8％) and margins clear in 21 patients (26.2％).Early phase enhancement was seen in 68 cases (85.0％),of which 45 cases (56.2％) had homogenous enhancement and 23 cases (28.8％) had heterogeneous enhancement.Late phase enhancement was seen in 73 cases (91.2％) and necrosis was hardly seen.Only two cases (2.5％) had liver metastasis without lymphatic metastasis.All the 20 colorectal cancer cases without schistosomiasis presented with single lesion,which was characterized by irregular bowel wall thickening and local ulcer.The average length of impaired intestine was (6.90±3.40) cm.No calcification was seen in these lesions.All cases had early phase lesion with remarkable heterogeneous enhancement with low density necrotic area.The late phase enhancement was decreased.Four cases (20.0％) had liver metastasis.Compared to cases without schistosomiasis,cases with schistosomiasis tended to have multifocal (x2=100,P=0.000) and longer lesion (t=45.506,P=0.000),and more calcification (x2=100,P=0.000).The early phase heterogeneous enhancement was less frequent (x2 =88,P=0.000) and late phase enhancement was more frequent (x2=100,P=0.000).The liver metastasis rate was significantly lower (x2 =8.688,P =0.014).Conclusions The CT imaging of colorectal cancer with schistosomiasis is characterized by calcifications inside the tumor with obscured margins and multiple intestinal segments involvement.Hematogenous metastasis and lymphatic metastasis are rarely seen.