The chalcone synthase (CHS) superfamily of the type III polyketide synthases (PKSs) generates backbones of a variety of plant secondary metabolites. Benzalacetone synthase (BAS) catalyzes a condensation reaction of decarboxylation between the substrates of 4-coumaric coenzyme A and malonyl coenzyme A to generate benzylidene acetone, whose derivatives are series of compounds with various biological activities. A BAS gene Pcpks2 and a bifunctional CHS/BAS PcPKSI were isolated from medicinal plant P. cuspidatum. Crystallographic and structure-based mutagenesis studies indicate that the functional diversity of the CHS-superfamily enzymes is principally derived from small modifications of the active site architecture. In order to obtain an understanding of the biosynthesis of polyketides in P. cuspidatum, which has been poorly described, as well as of its activation mechanism, PcPKS2 was overexpressed in Escherichia coli as a C-terminally poly-His-tagged fusion protein, purified to homogeneity and crystallized, which is helpful for the clarification of the catalytic mechanism of the enzyme and lays the foundation for its genetic engineering manipulation.
Butanones ; Catalytic Domain ; Crystallization ; Fallopia japonica ; enzymology ; Polyketide Synthases ; genetics ; metabolism

Objective To develop an oligonucleotide array to detect single nucleotide mutations in 23S rRNA gene.Methods Primers and probes targeting A2142G.A2143G and C2182T mutations in 23S rRNA gene were designed tp develop an oligonucleotide array.Samples were performed by an asymmetric PCR and the PCR products were hybridized with the specific DNA microarray chips.Non fluorescence-labeled PCR products were cloned into T vectors.The results of oligonucleofide array were confirmed by direct DNA sequencing and evaluated by minimal inhibitory concentration (MIC).Results The results obtained from oligonucleotide microarray were identical to those of direct sequencing.In 54 Helicobacter pylori samples,oligonucleotide microarray indicated that no A-to-C transition at 2142 was found,and the mutant rate of A2143G was 11.11 % (6/54),the mutant rate of C2182T was 12.96% (7/54).A2143C,A2143T,C2182A and C2182G mutations were not found.The other specimens were wild-type.All the above results were the same as that of MIC tests.Conclusions The oligonucleofide microarray is a reliable and accurate genotyping assay for clarithromycin-resistance of Helieobaeter pylofi.It is high-throughput screening method for gastric mucosa and improve the application of strategy for personalized therapy.

Objective To understand the psychological state , labor and childbirth knowledge awareness in hospitalized pregnant women with 37 to 41 weeks gestation and effect on rate of cesarean section , and to put forward the corresponding countermeasures .Methods Two hundred hospitalized pregnant women were chosen and divided into the labor group in which the pregnant women felt contractions regularly and the cervical dilatation was more than 1 cm and the no labor group in which the pregnant women were threatened labor or without, each with 100 cases.The self-rating anxiety scale (SAS), the self-rating depression scale (SDS), and the questionnaire on the knowledge of labor and delivery were used in two groups , and the delivery outcome was investigated.Results The scores of SAS and SDS were respectively (49.21 ±6.09), (58.97 ±6.43) in the no labor group, and were higher than (42.96 ±5.61), (49.95 ±6.02) in the labor group, and the differences were statistically significant (t=3.074, 4.146, respectively;P<0.01).The incidence rates of anxiety and depression in the no labor group were higher than those in the labor group , and the differences were statistically significant (χ2 =7.14, 5.64, respectively;P<0.05).The qualification rate of mastering the knowledge of labor and delivery was 63.0%in the labor group, and was 27.0%in the no labor group, and the difference was statistically significant (χ2 =26.18, P<0.01).The cases of cesarean section were 65 in the no labor group including 44 cases cesarean section induced by social factors , and were 19 in the labor group including 8 cases cesarean section induced by social factors , and the difference was found in the rate of cesarean section in two groups (χ2 =43.44, P <0.01 ).Two groups of pregnant women in labor oxytocin utilization comparative difference was statistically significant (P<0.01).Conclusions The main causes of early admission in the without labor pregnant women are respectively the psychological state before labor and the awareness of the knowledge of labor and delivery .The situation of the psychological state before labor and the awareness of the knowledge of labor and delivery in the no labor group are worse than that in the labor group , and the psychological health during pregnancy should be strengthened , and the propaganda and education of the knowledge of labor and delivery should be increased .

Objective To evaluate the bacteriostatic effect of recombinant human lactoferrin(rhLF) on Helicobacter(H .) py‐lori and its influence on CagA ,Ure and gastric mucosal IL‐8 .Methods The minimum inhibitory concentration(MIC)and the influ‐ence of different drug concentrations on the proliferation of H .pylori were detected .The effects of rhLF on the mRNA and protein expressions of CagA and Ure in H .pylori were detected by RT‐PCR and Western blot ,respectively .The animal study :Balb/c mice were adopted and assigned randomly into four groups ,including the standard triple+rhLF(group A) ,rhLF(group B) ,standard tri‐ple(group C) and normal saline(group D) .The histopathological HE staining was used to observe the gastric inflammation and ELISA was used to detect the IL‐8 level of gastric tissue in each group .Results MIC was 0 .5 mg/mL ,moreover rhLF inhibited the bacterial growth and proliferation with a concentration‐dependent manner .rhLF could reduce the expression of H .pylori major viru‐lence factor CagA ,mRNA and protein of Ure .Comparing the group A with the group B ,C and D ,the gastric mucosal inflammation score and the IL‐8 levels of gastric tissue homogenates had statistically significant differences(P<0 .05) .Conclusion rhLF inhibits the growth and proliferation of H .pylori ,moreover inhibit the expression of major virulence factor CagA in H .pylori ,mRNA and protein of Ure in different degrees ,weakens its pathogenicity ,meanwhile reduces the IL‐8 level in mice gastric mucosa ,and allevi‐ates H .pylori related gastric mucosal inflammatory response .

OBJECTIVETo investigate the association of SNP of CD40 gene and its serum levels with ischemic stroke (IS).

METHODSA total of 202 IS patients from a hospital of Baise city were enrolled in case group from May 2013 to November 2014. At the same time, 109 healthy people who had physical check-ups in the outpatient department at the same hospital were enrolled in the control group. All participants were from Guangxi Zhuang Autonomous Region and unrelated to each other. 3 ml venous blood were collected on the premise of informed consent. The single nucleotide polymorphisms of CD40 gene rs1883832 C/T, rs13040307 C/T, rs752118 C/T and rs3765459 G/A were analyzed using a Snapshot SNP genotyping assays, and the serum levels of CD40 were tested by ELISA. t-test was used to compare the serum levels of CD40 between the case and control group, and the genotypes at different locuses in case group; χ(2) test was used to compare the distribution differences of the CD40 gene locuses in different genotypes and allele between the case group and the control group; alleles was established as independent variables, the occurrence of the IS as dependent variable, and expressed relative risk with OR (95%CI) value.

RESULTSIn the case group, the frequency of CC, CT and TT genotypes in CD40 gene rs1883832 C/T were 21.78% (44/202), 49.51% (100/202) and 28.71% (58/202), respectively, and 33.17% (66/199), 48.74% (97/199), 18.09% (36/199) in the control group, respectively, the differences between the two groups was significant (χ(2)=9.57, P=0.008). The CD40 serum levels were (62.7 ± 24.5) pg/ml in the case group, which was higher than that in the control group (45.3 ± 17.2) pg/ml (t=8.97, P<0.001). The serum levels of TT and CT genotypes in CD40 gene were (65.9 ± 26.3) and (64.3 ± 25.9) pg/ml, respectively, and the differences were significant when comparing with CC genotype (t equaled 5.34 and 5.03, respectively, P<0.001). The risk of developing IS was 1.56 times higher in 1883832 T allele carriers than that in rs1883832 C allele carriers (OR=1.56, 95% CI: 1.18-2.06); Combined genotype analysis displayed that CD40 gene rs1883832 C/T, rs13040307 C/T, rs752118 C/T and rs3765459 G/A polymorphisms showed strong linkage disequilibrium, the case group TCCA haplotype was tested to be associated with a significantly increased risk of IS as compared with that in the control group(OR=2.49; 95%CI: 1.13-5.48).

CONCLUSIONCD40 gene rs1883832 C/T polymorphism and its TCCA haplotype were possibly associated with ischemic stroke, and the susceptibility gene for ischemic stroke may be rs1883832 T allele.

Alleles ; CD40 Antigens ; blood ; genetics ; Case-Control Studies ; Cell Differentiation ; China ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Polymorphism, Single Nucleotide ; Stroke ; blood ; genetics

Objective Studies show that women infected with human papillomavirus (HPV)induced cervicitis,cervical intraepithelial neoplasia (CIN) and on to develop cervical cancer their cervical mucosal exfoliated cells telomerase activity changes,to explore its clinical value in screening and early diagnosis and treatment of female patients with cervical cancer.Methods Elections of HPV testing negative control group 26 cases,HPV-positive group 28 cases,HPV-positive cervicitis group of 27 cases,HPV-positive CIN group 26 cases,24 cases of cervical cancer,HPV-negative cervicitis group 26 cases,HPV-negative CIN group of 23 cases of seven test group,cervical mucosa cytology specimens collected in 293 cells as positive standards for real-time fluorescence quantitative detection of telomerase TRAP Quantitative value of telomerase activity.Results Normal control group,ranging from 0 to 3.15,average 0.36; HPV-positive group,ranging from 0 to 21.6,with an average 1.98,＞ 10,3 cases; HPV-positive cervicitis group,ranging from 0 to 71.5,with an average 4.88,＞ 10,4 cases; HPV-positive CIN group,ranging from0 to 407,with an average 20.8,＞ 10,5 cases; cancer group,the range 1.2-5.5 × 106,median 346,where ＜ 10.4 cases; HPV-negative cervicitis group,ranging from 0 to 66.6,with an average 4.19,＞ 10,3 cases; HPV-negative CIN group,ranging from 0 to 158,with an average 14.2.Statistical analysis of cervical cancer group and the other groups are statistically significant (P ＜0.01),HPV-positive CIN group compared with the control group was statistically significant (P ＜ 0.05) the rest of the comparison between two groups was not statistically significance (P ＞ 0.05).Conclusions Women infected with HPV and cause cervicitis,CIN stage,the cervical mucosa telomerase activity gradually increased and in cervical cancer stage increased the most significant.And therefore take cells exfoliated cervical mucosa quantitative detection of telomerase activity can be used as effective method in screening and early diagnosis for cervical cancer in women.

Objective:To investigate the influences of SCN3A gene polymorphism(c.905A>G/p.N302S and c. 1441C>T/p.L481L) on the efficacy of valproic acid sodium in the treatment of Zhuangzu epilepsies.Methods:Using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)technique and the way of direct sequence, the SCN3A gene c. 905A>G/p.N302S and c. 1441C>T/p.L481L genotypes in peripheral blood were detected in 244 epileptic patients (85 cases in effective group and 139 cases of ineffective group) in the standardized treatment of valproic acid sodium.The blood concentration of valproic acid sodium was detected by LC-MS.Evaluating the correlation between the genotype and alleles of two groups of patients and the efficacy of valproic acid sodium and analyzing the difference of valproic acid sodium's blood concentration between different genotypes.The linkage disequilibrium of c. 905A>G/p.N302S and c. 1441C>T/p.L481L were analyzed by software SHEsis.Results:The allele and genotype distribution in c. 905A>G/p.N302S loci between effective group(A, G allele: 50.6%, 49.4%, AA, AG, GG genotype: 27.1%, 47.1%, 25.8%) and ineffective group(A, G allele: 37.4%, 62.6%, AA, AG, GG genotype: 16.6%, 41.7%, 41.7%) had statistically significant difference(χ 2=7.501, P=0.006; χ 2=7.907, P=0.019). There was no significant difference in allele and genotype distribution of c. 1441C>T/p.L481L loci between effective group(C, T allele: 47.1%, 52.9%, CC, CT, TT genotype: 23.5%, 47.1%, 29.4%) and ineffective group(C, T allele: 38.8%, 61.2%, CC, CT, TT genotype: 18.7%, 40.3%, 41.0%)(χ 2=2.920, P=0.088; χ 2=3.099, P=0.212). Compared with the AA + AG genotype, the GG genotype at c. 905A>G/p.N302S locus significantly reduced the efficacy of valproic acid sodium ( OR=2.051, 95% CI=1.136-3.703). Compared with genotypes AA+ AG, there were no significant differences in blood concentration of genotype GG of c. 905A>G/p.N302S ( t=3.256, P=0.137). Compared with genotypes CC+ CT, there were no significant differences in blood concentration of genotype TT of c. 1441C>T/p.L481L( t=4.628, P=0.082). c.905A>G/p.N302S and c. 1441C>T/p.L481L were without linkage disequilibrium. Conclusion:These results suggest that the single nucleotide polymorphisms of c. 905A>G/p.N302S in SCN3A genes may play a role in the resistivity of valproic acid sodium in Zhuangzu epilepsies.

Objective:To research the efficacy and safety of robotic thyroidectomy and bilateral modified radical neck dissection through bilateral axillo-breast approach (BABA) .Methods:We retrospectively analyzed the clinical data of 37 patients with thyroid cancer who received bilateral modified radical neck dissection through the BABA at the Department of Thyroid and Breast Surgery of the 960th Hospital of the People’s Liberation Army from Jan. 2014 to Jan. 2023. There were 24 females and 13 males, and the average age of the patients was (33,22±10.53) years old. The tumor diameter, number of lymph node dissection and metastasis in the central and lateral regions, average operation time, average hospital stay, complications, and aesthetic score were recorded. SPSS 25.0 software was used for statistical analysis, and the measurement data was calculated using mean ± standard deviation ( ± s), and the counting data was expressed in percentages and numbers. Results:A total of 37 thyroid cancer patients underwent robotic bilateral regional lymph node dissection. The 37 patients received total thyroidectomy, bilateral central compartment and cervical lateral regional lymph node dissection. All the pathological types were papillary carcinoma, with a maximal tumor diameter of (1.47±0.85) cm. The average number of central lymph nodes dissected was 19.46±8.84, and there were (10.24±5.95) metastases; The average number of lymph nodes removed from the bilateral cervical region was 38.92±14.21, and there were (7.92±5.84) metastases. The average operation time was (288.05±77.09) min, the average length of stay in the hospital was (10.76±3.92) days, and the average length of stay in the hospital following surgery was (8.03±2.08) days. These patients had no permanent hypoparathyroidism, permanent recurrent laryngeal nerve palsy, infection, accessory nerve injury and phrenic nerve injury after operation. Transient hypoparathyroidism occurred in 15 patients, transient recurrent laryngeal nerve palsy occurred in 1 patient, and chyle leak occurred in 2 patients. One month after surgery, the aesthetic score was 9.51±0.69.Two patients were found lymph node metastases during the (27.81±15.10) months of follow-up, and received robotic cervical lymph node dissection with BABA.Conclusion:For carefully chosen thyroid cancer patients with bilateral lateral cervical region lymph node metastases, robotic bilateral cervical lymph node regional dissection via BABA is safe and feasible, and good cosmetic results can be obtained.

Objective To investigate the current situation of competency and professional activities of pediatric rehabilitation thera-pists in Jiangsu,Shanghai and Zhejiang. Methods A questionnaire survey was conducted on 168 pediatric rehabilitation therapists in Jiangsu,Shanghai and Zheji-ang,from December,2022 to February,2023,and the general information and the competency were obtained.A total of 35 pediatric rehabilitation therapists were interviewed one-on-one,from October,2022 to January,2023,using thematic framework. Results A total of 162 effective questionnaires were collected.The level of competency of pediatric rehabilitation thera-pists was(3.44±0.87);and it was higher in professional literacy(3.90±0.77)and practice(3.83±0.77),and lower in management and leadership(2.92±1.44),and research(2.14±1.64).Latent class analysis showed that the thera-pists could be divided into four categories,of which the"medium competency"accounted for the most proportion(34.57%),and"low learning and research"was the least(8.64%).The interviews identified problems of compe-tency such as maintaining communication with children's families and managing self-professional development. Conclusion Self-reported rehabilitation competency levels of pediatric rehabilitation therapists in three provinces and cit-ies are above three points in all four dimensions,except for the management and leadership dimension,where the score is below three points.

Nonalcoholic fatty liver disease(NAFLD)is the most common chronic liver disease,with a global prevalence of approximately 30.05% to 32.4% .It is closely associated with various other diseases.In recent years,microRNAs(miRNAs)have played a crucial role as non-invasive biomarkers in understanding the pathogenesis and diagnosis of NAFLD.miRNAs play significant roles in both lipid metabolism and insulin resistance,exerting specific regulatory functions in the development and progression of NAFLD.miRNAs are small RNA molecules that regulate the gene expression and protein synthesis by controlling the transcription and translation of target genes.This article provides a comprehensive overview of the roles and mechanisms of miRNAs in lipid metabolism,insulin resistance,and the occurrence and development of NAFLD.