Objective: Clone, express and purify human recombinant calreticulin (CRT). Methods: Human CRT cDNA was amplified from total RNA of human lung cancer cell line A549 cells by RT-PCR. Then, PCR product was subcloned into prokaryotic expression vector pET-15b. After sequencing, this recombinant plasmid was transformed into E.coli. Rossetta. Recombinant CRT was expressed in host cells by IPTG induction. Resulted protein was purified by Ni-NTA resin under denature condition and dialyzed to recover its native structure. SDS-PAGE and Western blot method were used to identify the expression and purification of reconbinant CRT. Results: Human CRT cDNA was cloned from total RNA of A549 cells. CRT prokaryotic expression vector pET-15b-crt was constructed. Reconbinant CRT was induced to express in E.coli and purified by Ni-NTA affinity chromatograph. Conclusion: A method for prokaryotic expression and purification of human recombinant CRT was successfully established. This method laid a foundation for the subsequent CRT research.

Flavonoids is one of the biggest families of the plant-derived secondary metabolites with structural diversity. Until now, over 10 000 kinds of flavonoids with distinct structures have been purified and identified from plants, and some of them possess a range of important pharmacological effects, such as anticancer, anti-inflammatory and so on. So far, a number of genes and enzymes responsible for the biosynthesis of flavonoids have been reported, especially, a great of progress has been achieved in the synthetic biology of flavonoids in the recent years. Herein, based upon a brief introduction on the biosynthesis of flavonoids, this review summarizes the research advances in synthetic biology of flavonoids in the past two decades (2001-2021), highlighting the cell factories construction of the representative flavonoids. And, a brief discussion and prospects of the relevant metabolic bottlenecks and optimizing strategies are proposed.

Objective To explore the clinical value of transumbilica single-port laparoscopic surgery in the treatment of bilateral varicocele.Methods From Jan. 2010 to Sep. 2010,42 patients with bilateral varicocele underwent laparoscopic high ligation of bilateral spermatic vein.Of these patients,20 were treated with transumbilica single-port laparoscopic surgery,22 with traditional laparoscopic surgery.The two groups of patients were compared for the parameters such as intraoperative blood loss,testicular artery preservation,operating time,time of activities after surgery,time of intestinal function recovery and hospitalization duration.The semen quality 1 year after the surgery was compared with that before treatment.At the out-patient re-examination at 1,3,6months and 1 year after the surgery,the incision,scrotum,spermatic cord,testis were checked for possible complications.The relief of discomfortness in the scrotum was also followed up.Results Both operation procedures were successful,without severe complications.In the single-port laparoscopic surgery produced blood lose ( [ 5 ± 1 ] ml vs.[ 5 ± 1 ] ml,t =- 0.452,P ＞ 0.05 ),the operating time ( [ 41 ± 7 ] min vs.[ 39 ± 3 ] min,t =0.686,P ＞ 0.05 ),postoperative intestinal function recovery time ( [ 11 + 1 ] h vs.[ 11 + 2 ] h,t =- 1.631,P ＞0.05 ) and postoperative hospital stay ( [ 3.1 + 0.7 ] d vs.[ 3.4 + 0.7 ] d,t =- 1.447,P ＞ 0.05 ) which were all comparable to that from conventional laparoscopic surgery.There was significant difference in the population using analgesics,single-port laparoscopic surgery vs.conventional laparoscopic surgery ( 1 case [ 5.0％ ] vs 7case [ 31.8％ ].The difference was statistically significant (x2 =4.886,P ＜ 0.05 ).The single-port laparoscopic surgery produced neglectable scar at the incision.All of the patients were questionaired for their satisfaction with the incision 1 year after the surgery.The difference was statistically significant (x2 =7.636,P ＜ 0.01 )Conclusion Single-port laparoscopic high ligation of bilateral spermatic vein produces comparable outcomes to that of conventional laparoscopic surgery,but it is a more microinvasive procedure producing good aesthetic appearance,representing the trend of laparoscopic technique.

In order to evaluate the moUuseicidal effect of 50% wettable power of niclosamide ethanolamine salt(WPN)combined with urea against Oncomelania snails in the field,4 g/m~2 WPN,4 g/m~2 WPN+20 g/m2 urea and 4 g/m~2 WPN+30 g/m~2 urea were used for mollusciciding with the spraying method.The results showed after 7 days,the mortality rates of snail were 74.43% for 4 g/m~2 WPN,90.32% for 4 g/m~2 WPN+20 g/m~2 urea and 94.83% for 4 g/m~2 WPN+30 g/m~2 urea,respectively.It is indicated that WPN combined with urea can improve the molluscieidal effect significantly.

OBJECTIVETo investigate the mechanism of transcription regulation of GLI3 gene in idiopathic congenital talipes equinovarus.

METHODSpGL3-Gli3 luciferase report vectors were constructed, and the activity of Gli3 promoter was explored. A P-Match software was used to analyze the sequence upstream of the transcription start site of rat Gli3 gene, which was subsequently verified with chromatin immunoprecipitation assay (CHIP) and electrophoretic mobility shift assay (EMSA). Expression of the Gli3 gene was analyzed in L6 cells transfected with Hoxd13 small interference RNA(siRNA) and Hoxd13 expression vectors.

RESULTSThe 5' region of rat Gli3 gene contains two potential binding sites for the Hoxd13 protein. CHIP and EMSA assays both confirmed that Hoxd13 can directly bind with site 2. As shown in L6 cells, expression of Gli3 may be enhanced with silencing of Hoxd13, whilst exogenous expression of Hoxd13 can down-regulate transcription of Gli3.

CONCLUSIONHoxd13 can directly regulate the expression of Gli3 gene through a Hoxd13 binding site in the limb of rat embryo.

Animals ; Base Sequence ; Clubfoot ; genetics ; Gene Expression Regulation ; Homeodomain Proteins ; genetics ; Kruppel-Like Transcription Factors ; genetics ; Molecular Sequence Data ; Rats ; Rats, Wistar ; Transcription Factors ; genetics ; Transcription, Genetic ; Zinc Finger Protein Gli3

Objective To investigate the clinical significance of external obturation with cotton ball in treatment of digestive tract lip shape fistula. Methods A prospevtive randomixed controllde study was conducted in 30 patients with digestive tract lip shape fistula admitted from January 2001 to November 2007,which were divided into experimental group ( 15 patienets) and control griup ( 15 patjients). The control group received traditional fistula mouth continuous drainage,with fastomg or part enteral nutrition and parenteral nutrition. The experimernal group received external obturatiion with cotton ball,with controling spills of oineteatinal fluid and resuming normal eating and activetes, The patients general information, fistuala mouth sixe, volume, weight, albumin, determinstic operation time, postoperative complications, hospital, signficantly reduced fistula, flow, postoperative compared between the two groups. Results Compared with the control group , significantly redced fistula, flow, postoperative complivations and cost, increased wdighe and albumin,as well as shortened operation time and hospital stay were found oin the experimetal grou. Conclusin External obturation with cotton ball is safe effective methde in treatment of digestive tract lip shape fistula, obviusly shortening the course of disease, redcing patients'cost and pain, and rapidly improving patients nutritional status.

Abstract: Objective To analyze the effect of nutrition package on the nutritional status and prevalence of children in rural areas of Hainan Province, and provide scientific basis and suggestions for further improving the nutritional and health status of children in this region. Methods　Four cities and counties were randomly selected as the intervention group, and four cities and counties matched with the intervention group in terms of population, economy, social culture, maternal and child health work foundation of township health centers, physical nutrition and health status were selected as the control group.With the combination of monitoring and prospective cohort study, infants in the intervention group and the control group were studied from June 1, 2020, and they were intervened for 12 months with supplementary food nutrition package. Before and after intervention, the nutrition and health status of infants aged 6-24 months in the intervention group and the control group were investigated to evaluate the nutritional and health effects of supplementary food nutrition package for infants aged 6-24 months in rural Hainan Province. Results　A total of 999 infants were investigated, including 427 in the intervention group and 572 in the control group. After 12 months of nutritional intervention, there was no significant difference in weight-for-age Z-score (WAZ) and height-for-age Z-score (HAZ) and weight-for-height Z-score (WHZ) between the intervention group and the control group (P>0.05). The rate of emaciation of the intervention group was 1.64%, which was significantly lower than 3.67% of the control group (P<0.05). There were no significant differences in the rate of growth retardation (2.81% and 3.32%, respectively) and underweight (0.47% and 1.92%, respectively) between the intervention group and the control group (P>0.05). The rate of respiratory infection and diarrhea in the intervention group were 9.13% and 1.17%, which were significantly lower than corresponding 23.25% and 3.15% in the control group (P<0.05). The hemoglobin of the intervention group and the control group were 117.24 g/L and 114.51 g/L respectively, and the rates of anemia were 11.11% and 22.84% respectively, the differences were statistically significant (P<0.05). Conclusions　The intervention of nutrition package in rural areas of Hainan Province has achieved the expected results, and supplementary food nutrition package has reduced the incidence of malnutrition and respiratory infection and diarrhea in recent two weeks in infants and anemia to a certain extent. We should attach great importance to the supplementary nutrition package for right-age children and promote the growth and health of children in rural areas through supplementary nutrition package, and continuously improve the nutrition and health level of children in Hainan Province.

OBJECTIVETo study the relationship between insertion/deletion (I/D) polymorphism of 287 bp in the 16th intron of angiotensin converting enzyme (ACE) and essential hypertension in children.

METHODSI/D polymorphism of 287 bp in the 16th intron of ACE was detected using PCR in 105 children with essential hypertension and 105 normal children as control group.

RESULTSThere was an I/D polymorphism in the 16th intron of ACE in the hypertension and the control groups: type II, type ID and type DD. The genotype frequencies of type DD, type ID and type II in the hypertension group were 30.5%, 47.6% and 21.9%, respectively. The genotype frequencies of type DD, type ID and type II in the control group were 14.3%, 46.7% and 39.1%, respectively. There were significant differences in the genotype frequencies of types DD and II between the two groups (P<0.01). The allele frequency of type D (54.3% vs 37.6%) was significantly higher in the hypertension group; in contrast, the allele frequency of type I (45.7% vs 62.4%) was significantly lower than in the control group (P<0.01).

CONCLUSIONSPolymorphism of type II, type ID and type DD exits in ACE. The deletion of 287 bp in the 16th intron of ACE might be associated with the occurrence of essential hypertension in children.

Adolescent ; Child ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Genetic

OBJECTIVETo investigate whether cyclooxygenase-1 (COX-1) haplotype is associated, with aspirin resistance.

METHODSThe participants were 431 old Chinese Han patients with cardiovascular and cerebrovascular diseases who took aspirin. The 59 patients with aspirin resistance (AR) by light transmittance aggregation acted as the cases; the 372 aspirin-sensitive patients were the controls. The relationships between AR and 6 single nucleotide polymorphisms (SNPs) in COX-1 gene. rs1888943 (8759C/T), rs1330344 (1676A/G), rs3842787 (exon2, 50C/T, p.Pro17Leu), rs5787 (exon 4, 323G/A, p. ARg108Gln), rs5789 (exon7, 709C/A, p. Leu237Met) and rs5794 (exonl0, 1330G/A, p.Va1481Ile) were investigated by the USA Sequenom high-throughput single nucleotide polymorphisms (SNP) genotyping systems.

RESULTSIn this case-control trial, the frequency of mutant CGCGCC-haplotype in case was 0.48 (57/118) and in control was 0.39 (286/742), which was significantly higher than that of the control group (P < or = 0.05).

CONCLUSIONCOX-1 haplotype is associated with aspirin resistance in old Chinese Han patients with cardio-cerebrovascular diseases, mutant CGCGCC-haplotype carriers of COX-1 has a significant significantly increased risk of AR.

Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Aspirin ; pharmacology ; Cardiovascular Diseases ; genetics ; Case-Control Studies ; Cerebrovascular Disorders ; genetics ; Cyclooxygenase 1 ; genetics ; Drug Resistance ; genetics ; Female ; Genotype ; Haplotypes ; Humans ; Male ; Polymorphism, Single Nucleotide

The purpose of the present study was to investigate the effect of meperidine on rat ventricular muscle. Cardiac function was assessed in Langendorff-perfused rat hearts and intracellular calcium level was recorded in enzymatically isolated rat ventricular myocytes using spectrofluorometric techniques. To explore the underlying mechanism, whole-cell configuration of patch-clamp technique was used to record L-type Ca(2+) current. The results showed that meperidine decreased the product of heart rate and left ventricular developed pressure (LVDP HR), maximal rate of the left ventricular pressure increase (LV +dP/dt(max)) and decrease (LV -dP/dt(max)), but increased left ventricular end-diastolic pressure in a dose-dependent manner (0-1000 micromol/L). Meperidine also produced a dose-dependent reduction in electrically induced [Ca(2+)](i) transient amplitude and an increase in diastolic [Ca(2+)](i) baseline level, but did not alter the caffeine (20 mmol/L) induced Ca(2+) release from intracellular ryanodine-sensitive Ca(2+) stores. Meperidine at 100 micromol/L inhibited L-type Ca(2+) current to 67.4 10.1% of control but did not affect the voltage dependency of activation and inactivation. The inhibitory effect of meperidine on Ca(2+) current could not be prevented by pretreatment with the opioid receptor antagonist naloxone. These data suggest that meperidine exerts a negative inotropic effect by inhibiting L-type Ca(2+) current. The lack of effect of naloxone implies that the action is independent of the opioid receptor.
Animals ; Calcium Channels, L-Type ; drug effects ; Depression, Chemical ; Dose-Response Relationship, Drug ; Heart Rate ; drug effects ; Male ; Meperidine ; pharmacology ; Myocardial Contraction ; drug effects ; Myocytes, Cardiac ; metabolism ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Ventricular Function, Left ; drug effects