Objective To obtain abundant human betacellulin(BTC) with biological activity. Methods The whole mature protein coding sequence of BTC gene was amplified by polymerase chain reaction(PCR) method applied to human pancreatic ?-cell tumors cDNA.The fragment was cloned into prokaryotic expression vector pET32a(+) plasmid.The recombinant plasmid was transformed into E.coli BL21 and the fusion protein was expressed under isopropyl-beta-D-thiogalactopyranoside(IPTG).The fusion protein was purified by Ni2+ affinity chromatography.SDS-PAGE and Western blot were employed to determine the expression and purification of the expected protein.BTC was added to culture NIH3T3 cells for 5 days,and cell proliferation was detected by MTT. Results Lots of fusion protein were produced,and the purified protein can stimulate the proliferation of NIH3T3 cells. Conclusion The human BTC can be successfully obtained from the pET32a(+) system with the biological activity of stimulating the proliferation of NIH3T3 cells.

Objective To construct eukaryotic expression vector of human pancreatic duodenal homeobox 1(PDX-1) gene,and to detect its expression in NIH3T3 cell lines. Methods The whole coding sequence of PDX-1 gene was amplified by polymerase chain reaction(PCR) from human pancreatic-cell tumors cDNA.The fragment was inserted into eukaryotic expression vector pcDNA3.1 plasmid.The recombinant plasmid was verified by double digestion and DNA sequencing.The expression of PDX-1 gene in NIH3T3 cells was assayed by Western blot. Results The length of specific fragment amplified by PCR was 852 bp,and the recombinant plasmid pcDNA3.1-PDX-1 showed two bands of 5.5 kb and 852 bp by digestion using respective restriction enzymes BamHⅠand EcoRⅠ.The sequence of PDX-1 gene was approved or confirmed by blasting to GenBank.It was suggested that PDX-1 gene had been cloned into pcDNA3.1 vector correctly.Western blot showed that PDX-1 gene was expressed,which was detected 24 h after pcDNA3.1-PDX-1 plasmid was transfected into NIH3T3 cells. Conclusion The recombinant eukaryotic expression vector pcDNA3.1-PDX-1 was successfully constructed and expressed in NIH3T3 cell lines.

Objective To investigate the prevalence rate of new-onset malignant tumors in type 2 diabetes mellitus patients .Methods Type 2 diabetes mellitus patients ( 1771 cases ) over 3 years since May 2006 in Xinhua Hospital were retrospectively recruited .The prevalence and distribution of different tumors were analyzed .Results ⑴Total prevalence of malignant tumors was 2.88%, and more common in male ( P <0.05 ) .The prevalence of different malignant tumors in human systems from high to low were digestive, urinary, and respiratory .⑵Age-dependent analysis from <50, 50~, 60~, 70~,and≥80 showed that the prevalence were 0.73%,1.93%,3.27%,3.58%, and 5.02% respectively .A significant difference was found in them ( P <0.05 ) .Conclusion The prevalence of new-onset malignancies is higher in type 2 diabetes mellitus , which is associated with age and sex .Tumor screening should be strongly recommended in such a group of the patients for their earlier diagnoses and treatments due to the un -typical clinical manifestations .

Objective To evaluate the efficacy of ultrasound?guided transversus abdominis plane ( TAP) block with different concentrations of ropivacaine for analgesia after cesarean section. Methods A total of 120 parturients, aged 24-31 yr, of American Society of Anesthesiologists physical statusⅠ or Ⅱ, weighing 64-73 kg, at 35 to 41 week gestation, scheduled for elective cesarean section, were randomly di?vided into 3 groups ( n= 40 each) using a random number table: 0?25% ropivacaine group ( group Ⅰ) , 0?20% ropivacaine group ( groupⅡ) , and 0?15% ropivacaine group ( groupⅢ) . A patient?controlled an?algesia pump was connected at the end of surgery, and the corresponding concentration of ropivacaine 1?5 mg∕kg was injected into the bilateral TAP under the guidance of ultrasound in each group. Visual analogue scale score was maintained ≤3. The number of attempt and the number of patients requiring rescue analge?sic were recorded at 48 h after surgery. The occurrence of TAP block?related complications was observed and recorded. Results Compared with groupⅢ, the number of attempt and the number of patients requi?ring rescue analgesic were significantly decreased in Ⅰ and Ⅱ groups ( P<0?05 ) . Compared with groupⅡ, the number of attempt and the number of patients requiring rescue analgesic were significantly de?creased in group Ⅰ (P<0?05). No TAP block?related complications were detected in the three groups. Conclusion Ultrasound?guided TAP block with 0?25% ropivacaine is helpful in improving the analgesic efficacy after cesarean section without serious complications.

Aim To study the effect of emodin on IL-8 secretion and NF-?B activation of HT-29 cells,and explore the molecular mechanism of emodin.Methods The cytotoxicity of emodin was assessed by WST;NF-?B activation was detected with co-focal microscopy by immunofluorescence;the production of IL-8 was investigated by ELISA.Results Emodin with the concentration of 10~80 ?mol?L-1 could decrease the mass production of IL-8 Secretion of HT-29 cells stimulated by IFN-?+LPS in a dose-dependent manner.Emodin with various concentrations could inhibit NF-?B activation dose-dependently.Conclusions Emodin inhibited IL-8 secretion and NF-?B activation of HT-29 cells stimulated by IFN-?+LPS.

Objective To investigate the effect of triiodothyronine on the expression of alpha subunit of C protein Co (Coa) gene in cultured rat cerebral cortex neurons. Methods Primary cultured neurons of rat were prepared from the cerebral cortex at embryonic day 19. The neurons were cultured in DMEM supplemented with 15% fetal calf serum. After 7 days, the neurons were cultured in different media; DMEM supplemented with 15% fetal calf serum (group A), DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group B), 0.5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group C) , 5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group D) and 50nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group E). The neurons were cultured for another 7 days and total RNA was extracted. Goa mRNA leves were measured by competitive RT-PCR. Results As compared with group A, Coa mRNA leves in group B, D and E were low (P 0.05). Conclusion T, has dual effects on the expression of Coa gene in cultured rat cerebral cortex neurons. T3 down-regulated Coa mRNA in cultured neuron. However, in the absence of T3, Goa mRNA was also very low. Thyroid hormone may exert their action on brain development by regulating the the expression of Goa gene.

Objective To evaluate the distribution of non-motor symptoms(NMS) and dominant factors impacting on the prevalence of NMS in Parkinson's disease(PD). Methods The NMS questionnaire(NMSQuest) was administered to 128 patients(PD group) and 128 healthy adults(control group).The distribution of items of NMS was analyzed between the two groups,and clinical characteristics were also collected to assess the dominant factors impacting on the prevalence of NMS. Results There were significant differences in the prevalence of NMS between the two groups(P

A thiosulfate-oxidizing bacterium TX was isolated from polysulfide wastewater treatment system.It was preliminarily identified as Halothiobacillus sp.according to its morphological,physiological proper-ties and similarity analysis of 16S rRNA gene sequence(GenBank accession No.EU871645).This bacte-rium was an obligate chemolithoautotrophic sulfur-oxidizing bacterium since it could utilize thiosulfate,elemental sulfur,tetrathionate,sulfide or sulfite as sole energy source but did not show growth on the me-dium with glucose,sucrose,fructose,lactose,mannose or yeast extract as sole carbon source.The optimal growth condition for thiosulfate oxidation was 30?C~35?C and pH 3.0~5.0 respectively.Significant reduc-tion in medium pH from 4.0 to 1.2 was observed due to oxidation of thiosulfate to sulfate during the batch culture incubation period.During the course of thiosulfate oxidation accumulation of tetrathionate was ob-served either in synthetic medium or in the mixed liquor from aeration tank of the polysulfide wastewater treatment system.This indicates that the "S4 intermediate(S4I)" pathway was responsible for thiosulfate oxidation in this bacterium.

Objective:To explore the application value of video-electroencephalogram(VEEG) monitoring in the diagnosis and localization of children with epilepsy.Methods:The clinical data of 310 children with clinical epilepsy symptoms diagnosed and treated in Tangshan Maternal and Child Health Care Hospital and Qilu Children′s Hospital Affiliated to Shandong University from May 2018 to April 2020 were analyzed. All children underwent routine EEG (REEG), VEEG monitoring, and positron emission computed tomography (PET-CT) for children undergoing surgery. REEG and VEEG was compared to monitor epileptic form discharges; the location of epileptic lesions was compared by different examination methods, and children were followed up after surgery.Results:Among the 310 children, 247 children were diagnosed with epilepsy and 63 children were suspected of epilepsy. The detection rate of VEEG epileptic form discharge was higher than that of REEG: 87.4% (216/247) vs. 45.7% (113/247), and the difference was statistically significant ( χ2 = 6.3042, P<0.05). Among 247 children with epilepsy, during the VEEG test, 81 children (32.8%) had clinical symptoms, of whom 65 had seizures and 16 had non-epileptic seizures. The detection rate of VEEG for epilepsy was higher than that of clinical seizures: 87.5% (216/247) vs. 32.8% (81/247), and the difference was statistically significant ( χ2 = 8.6148, P<0.05). Among the 247 children with epilepsy diagnosed, 144 children had epilepsy syndrome, temporal lobe epilepsy accounted for 54.17% (78/144), and frontal lobe epilepsy accounted for 34.2% (50/144). The accuracy of preoperative VEEG localization of epilepsy lesions in 50 children undergoing surgery was higher than that of PET-CT and REEG: 80.0% (40/50) vs. 56.0% (28/50), 54.0% (27/50), and the differences were statistical significance ( χ2 = 5.3014, 5.6031, P<0.05). Postoperative follow-up of 50 children who had underwent surgery showed that 50.0% (25/50) of the children had no seizures, and 38.0% (19/50) of the children had significant improvements in postoperative seizure time, seizure period, and clinical manifestations. 12.0% (6/50) of the children had no significant improvement in clinical symptoms. Conclusions:VEEG can improve the detection rate of epileptic form discharges, and provide an important basis for clinical diagnosis of epilepsy, and have important value in locating epilepsy lesions before surgery.

Objective To investigate the relationship between the serum apolipoprotein B (ApoB)/ ApoA-Ⅰ ratio and intracranial artery stenosis in young patients with ischemic stroke.Methods Patients with ischemic stroke aged from 18 to 45 were enrolled in the study.Brain CT angiography was used to evaluate the degree of intracranial artery stenosis,and the concentrations of serum total cholesterol,triglyceride,highdensity lipoprotein cholesterol,low-density lipoprotein cholesterol,ApoA-Ⅰ,and ApoB were detected.The ratio of ApoB/ApoA-Ⅰ was calculated.The Demographic and clinical characteristics of the intracranial artery stenosis group and the non-intracranial artery stenosis group were compared.Multivariate logistic regression analysis was used to identify the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke.Results A total of 161 young patients with ischemic stroke were enrolled,including 89 in the intracranial artery stenosis group and 72 in the non-intracranial artery stenosis group.The constituent ratios of diabetes mellitus (20.2％ vs.6.9％;x2 =4.641,P =0.032),smoking (47.5％ vs.15.2％;x2 =15.121,P=0.001),hyperlipidermia (56.1％ vs.48.6％;x2 =4.197,P=0.040),as well as the radios in serum high-density lipoprotein cholesterol (1.29 ± 0.30 mmol/L vs.1.65 ± 0.34 mmol/L;t =7.131,P=0.002),ApoA-Ⅰ (1.49 ± 0.65 g/L vs.1.63 ± 0.23 g/L;t =2.751,P =0.001),ApoB (1.49 ± 0.65 g/L vs.1.63±0.23 g/L;t=2.751,P=0.001),and ApoB/ApoA-Ⅰ ratio (1.49±0.65 vs.1.63± 0.23;t =2.751,P=0.001) had significant differences between the two groups.Multivariate logistic regression analysis showed that diabetes (odds ratio [OR] 3.052,95％ confidence interval [CI] 1.186-7.856;P =0.021),smoking (OR 2.997,95％ Cl 1.456-6.172;P =0.003),hyperlipidemia (OR 4.745,95％ CI 2.108-10.668;P =0.001),ApoB (OR 4.861,95％ CI 3.029-7.802;P=0.001),and ApoB/ ApoA-Ⅰ ratio (OR 5.684,95％ CI 2.215-14.584;P=0.002) were the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke,while HDL-C (OR 0.561,95％ CI 0.354-0.888;P=0.014) and ApoA-Ⅰ (OR 0.065,95％ CI 0.010-0.409;P=0.004) were the independent protective factors.After adjustment for hypertension,diabetes,smoking,hyperlipidemia,HDL-C,ApoA-Ⅰ,and ApoB,ApoB/ApoA-Ⅰ ratio was still an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke (each increase of 1 standard deviation,OR 4.255,95％ CI 2.348-7.711;P=0.001).Conclusion ApoB/ApoA-Ⅰ ratio is an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke.