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 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: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: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

BACKGROUND: The content of aluminium generally increases in the cerebral cells of patients with senile dementia. Aluminium poisoning in brain has inner link with senile dementia. Naloxone is the specific antagonist of opioid receptor, which can be applied in the treatment of senile dementia according to foreign reports.OBJECTIVE: To investigate protective effects of naloxone on aluminium trichloride-induced memory impairment of senescence-accelerated mice and its mechanism.DESIGN: Randomized controlled trial.SETTING: Jilin Medical College.MATERIALS: The experiment was completed in Laboratory of Pharmacology of Jilin Medical College (formerly the Jilin Military Medical College) from February 2001 to February 2003. A total of 100 healthy adult Kunming mice were selected and randomly divided into 5 groups: control group, model group, naloxone 0.1 mg/kg group, naloxone 0.3 mg/kg group and naloxone 0.9 mg/kg group, with 20 in each group. Except the control group, subcutaneous injection with 70 mg/kg aluminium trichloride was given to the mouse in each group once a day for continuous 7 days; besides this, intraperitoneal injection with 0.1, 0.3, 0.9 mg/kg naloxone was given to the mouse in naloxone groups and the same amount of physiological saline was given to the mouse in the control group.METHODS: The methods of jumping stand and escaping dark were conducted to detect learning ability and memory of mice. Meanwhile, the content of malondialdhehyde in liver and mono-amine oxidase B in brain of mice were also detect.MAIN OUTCOME MEASURES: ① Results of jumping stand experiment of aluminium trichloride-induced model of senescence. ② Results of escaping dark experiment of aluminium trichloride-induced model of senescence. ③ Comparison of malondialdhehyde and mono-amine oxidase B among each group.RESULTS: ① Results of jumping stand experiment of aluminium trichloride-induced model of senescence: Compared withmodel group, the frequency of electric shocks suffered by aluminium trichloride mice and the amount of suffered animals within 5 minutes significantly decreased in control group and naloxone 0.1, 0.3, 0.9 mg/kg groups. The experiment was repeated 24 hours later, naloxone could significantly prolong the latency of the mouse jumping down from the platform for the first time (P ＜ 0.01).Meanwhile, naloxone could decrease the amount and the frequency of mist&es of aluminium trichloride mouse within 3 minutes (P ＜ 0.01). ② Results of escaping dark experiment of aluminium trichloride-induced model of senescence: Compared with model group, the latency of aluminium trichloride mouse entering dark box was significantly prolonged and the frequency of electric shocks suffered by aluminium trichloride mice obviously increased in control group and naloxone 0.1, 0.3, 0.9 mg/kg groups (P＜0.01). ③ Comparison of malondialdhehyde and mono-amine oxidase B among each group: Malondialdhehyde was more in model group than in naloxone groups (P＜0.01). Mono-amine oxidase B was more in model group than in the other groups (P＜0.01).CONCLUSION: Naloxone has protective effects on aluminium trichlorideinduced memory impairment of senescence-accelerated mice, which can improve learning ability and memory. The mechanism is probably relevant to the effects of decrease of mono-amine oxidase B and elimination of lipid peroxide.

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 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