AIM:To investigate the effects of Balanophora involucrata Hook.f.(BIH)extracts on bile acid metabolism and liver injury in mice with metabolic dysfunction-associated fatty liver disease(MAFLD)through the gut mi-crobiota-farnesoid X receptor(FXR)axis,and to explore the underlying mechanisms.METHODS:Forty C57BL mice were randomly divided into control group,MAFLD model group,medium-dose BIH group,and high-dose BIH group.The mice in control group received a regular diet,while those in other groups were fed with high-fat diet for 12 weeks to induce MAFLD.The mice in medium-and high-dose BIH groups received 0.598 and 0.299 g/kg BIH solution,respectively,while those in control and MAFLD groups received an equivalent volume of normal saline.Serum levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were measured using an automatic biochemical analyzer.Liver morphology,steatosis and fibrosis were assessed by HE,oil red O and Masson staining.Levels of TC,tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in liver tissues,and bile acids in serum and ileum tissues were measured by ELISA.Protein expression of FXR and fibroblast growth factor 15(FGF15)in ileum tissues,and FXR,small heterodimer partner(SHP)and cholesterol 7α-hydroxylase(CYP7A1)in liver tissues were analyzed by Western blot.Intestinal microbiota changes were assessed by 16S rRNA gene sequencing.RESULTS:(1)The MAFLD mice exhibited increased serum TC,TG,LDL-C and bile acid levels,liver TC,TNF-α and IL-6 levels,and lipid deposition.However,BIH intervention improved these factors and increased FXR and SHP pro-teins,but decreased CYP7A1 expression in the liver.The protein levels FXR and FGF15 in the ileum were also elevated.(2)Intestinal flora analysis demonstrated that BIH intervention improved the abundance and diversity of intestinal flora in MAFLD mice.Specifically,there was an increase in Bacteroidetes/Firmicutes ratio and a decrease in Proteobacteria and Verrucomicrobia.At the genus level,abundance of Duncaniella,Muribaculum and Paramuribaculum increased,while He-licobacter decreased.CONCLUSION:Treatment with BIH regulates intestinal flora,decreases FXR levels,enhances CYP7A1 expression,promotes bile acid synthesis,reduces hepatic cholesterol accumulation,and attenuates liver steato-sis and inflammation in MAFLD mice,indicating potential therapeutic effects.

【Objective】 To analyze the commonality and characteristics between voluntary blood donors and hematopoietic stem cell donors in this region, and explore the potential for integration and development between China Marrow Donors Program (CMDP) and voluntary blood donors, especially platelet donor databases, so as to improve recruitment success rate and inventory rate. 【Methods】 The database modeling and comparison methods were used to screen and stratify the matching and integration degree between the voluntary blood donors in recent 10 years and the marrow donors in the Shaanxi Branch of CMDP. The frequencies of HLA-A,-B alleles, HPA alleles and haplotypes were calculated with Arlequin 3. 5. 2. 2 software, and the matching probability of different platelet donor reserve pools was conducted according to the phenotypic frequencies. 【Results】 Among the voluntary donors with known HLA genotypes in this region, according to their blood donation behavior,the active blood donors excavated were divided into the first, second, third and fourth echelons of platelet donor reserve pools, with 696, 2 752, 9 092 and 12 028 donors, respectively. The first echelon had the highest proportion of 10-50 times of platelet donations and 10-20 times of whole blood donations, with 13.65% and 26.01%, respectively. The second echelon had 10-20 times of whole blood donations and 10-50 times of platelet donations, accounted for 15.04% and 1.38%, respectively, which were significantly different from other echelons' blood donation characteristics (P<0.05). With a database size of the existing platelet donor bank adding the first and second echelons (n=4 955), there was a 69.02% probability of matching at least one donor with matching HLA-A-B phenotype. When considering the matching ABO and HPA phenotypes, the probability of finding at least one donor with fully matching HLA, HPA and ABO isotype (type B as an example) was 48. 73%. 【Conclusion】 The three groups of whole blood donation, apheresis platelet donation and marrow donation in Xi'an area have a large cross-distribution. Compared with expanding the storage capacity from scratch, the active blood donors in CMDP database are the largest back-up force of platelet donors. While expanding the effective storage capacity, it can minimize the cost of building platelet donor bank and the demand for resources.

Objective:To construct a quality evaluation index system for postoperative rehabilitation in children with developmental dislocation of the hip (DDH) .Methods:From August to November 2021, a quality evaluation index system for postoperative rehabilitation in children with DDH was established through literature review, group discussion, and Delphi expert consultation. The analytic hierarchy process was used to determine the weights of various indicators. The positive coefficient of experts was expressed by the effective response rate of the questionnaire. The authority level of experts was represented by the authority coefficient ( Cr) , which was determined by the expert judgment coefficient ( Ca) and the expert familiarity level ( Cs) . The degree of coordination of expert opinions was represented by the Kendall harmony coefficient (Kendall's W) . The degree of concentration of expert opinions was represented by the average score of importance assignment, coefficient of variation, and full score rate. Results:The effective response rates of the two rounds of expert consultation questionnaires were 90.00% and 100.00%, respectively. The Cr of experts in two rounds were 0.785 and 0.815, and the Kendall harmony coefficients were 0.098-0.193 and 0.147-0.212 ( P<0.05) . In the second round of expert consultation, the average scores of importance assignment for all levels of indicators were≥4.0, with coefficients of variation of 0-0.24 and a full score rate of ≥50%. The final quality evaluation index system for postoperative rehabilitation in children with DDH included 3 first-level indicators, 9 second-level indicators, and 37 third-level indicators. In the consistency test results of each indicator at all levels, all of the CI were <0.001, and all of the CR were<0.001. Conclusions:The construction process of the quality evaluation index system for postoperative rehabilitation in children with DDH is scientific, and the weight distribution of the indicators is reasonable, which can be used as an effective tool for evaluating the postoperative rehabilitation quality of DDH children.

【Objective】 To evaluate the appropriate optimal capacity and matching probability of the platelet donor database with known HLA/HPA genotype in Shaanxi aera, and provide data support for subsequent construction, maintenance and application of the local platelet donor database. 【Methods】 A total of 11 755 individuals from the Shaanxi Branch of China Marrow Donor Program, 401 and 249 unrelated random platelet donors in Shaanxi aera were enrolled to the population study of HLA-A, -B polymorphisms, HPA genotyping and CD36 antigen expression, respectively. The frequencies of HLA-A, -B alleles, HPA alleles and haplotypes were calculated with Arlequin 3. 5. 2. 2 software; matching probability and capacity evaluation of platelet donor database was conducted according to the phenotypic frequencies. 【Results】 The population genetic and phenotypic polymorphisms data of HLA-A, -B and HPA1-6, 10, 15, 21 in Shaanxi aera were obtained. The frequency of CD36 type Ⅰ or Ⅱ deficiency was 0.40%(1/249). According to the subsequent calculating and deriving, with a database size of 194 donors, the patient having approximate 95% probability could achieve matching of HPA1-6, 10, 15, 21 genotype. With a database size of 1500 donors, there is a 95% probability of matching at least one donor with HLA-A-B phenotype frequency >0.002 or haplotype frequency >0.001; meanwhile, the probability of matching a cross-reactive group donor should be 44.95%-97.57%. Based on database size of 8 856 and 15 033, the probabilities of matching HLA-A, -B phenotype were about 80% and 90%, respectively. 【Conclusion】 The differences in the distribution of HLA/HPA polymorphism in different regions make the establishment mode and optimal capacity of platelet donor database different. It is necessary to apply a variety of platelet matching transfusion strategies to expand the range of donor selection, thereby effectively reducing the database construction cost and resource requirements.

Objective:To explore the application effect of family-centered empowerment model (FCEM) in rehabilitation nursing of children with femoral shaft fractures treated by skin traction.Methods:A retrospective case-control study was conducted to analyze the clinical data of 107 children with femoral shaft fractures treated with skin traction in Anhui Children′s Hospital from April 2019 to October 2021, including 67 males and 40 females; aged 1.5-5.0 years [2.8(2.0, 3.5)years]. Fracture types included oblique fracture ( n=50), comminuted fracture ( n=26), transverse fracture ( n=14) and spiral fracture ( n=17). FCEM nursing was implemented in 55 patients (family empowerment group) and routine home rehabilitation nursing guidance was performed in 52 patients (routine nursing group). The face, legs, activity, cry, consolability (FLACC) behavioral score on admission and at days 1, 3 and 7 after skin traction, treatment compliance rate at 1 month after skin traction, complication rate at 1 month after skin traction, length of hospital stay and fracture healing time were compared between the two groups. Results:All patients were followed up for 12-18 weeks [13.4(12.7, 13.9)weeks]. There was no significant difference in FLACC behavioral score between the two groups on admission ( P>0.05). The FLACC behavioral score was (4.0±0.7)points,(3.4±0.6)points and (2.4±0.6)points in family empowerment group at days 1, 3 and 7 after skin traction, lower than (4.8±0.7)points, (3.9±0.8)points and (3.3±0.5)points in routine nursing groups (all Ρ<0.01). One month after skin traction, treatment compliance was excellent in 51 patients, good in 4, poor in 0, with treatment compliance rate of 92.7% (51/55) in family empowerment group, and was excellent in 40 patients, good in 9, poor in 3, with treatment compliance rate of 76.9% (40/52) in routine nursing group ( Ρ<0.05). The complication rate was 5.4% (3/55) in family empowerment group, lower than 21.2% (11/52) in routine nursing group ( Ρ<0.05). The length of hospital stay and fracture healing time were (15.6±0.3)days and (7.1±0.8)weeks in family empowerment group, shorter than (16.8±0.3)days and (8.2±0.4)weeks in routine nursing group (all Ρ<0.01). Conclusion:For children with femoral shaft fractures treated by skin traction, nursing using FCEM can alleviate pain, improve treatment compliance, reduce complication rate, reduce hospital stay and shorten fracture healing time.

To construct TeI3c/4c-based and temperature-inducible gene inactivation system (Thermotargetron) and to apply it to gene inactivation of mesophilic bacteria. The subunit of flagellum (fliC) and C4 dicarboxylate orotate:H⁺ symporter (dctA) genes were chosen as targets in the genome of Escherichia coli HMS174 (DE3) strain. According to recognition roles of TeI3c/4c intron, the fliC489a, fliC828s, fliC1038s and dctA2a sites were chosen as target sites. Gene-targeting plasmids were constructed based on pHK-TT1A by using overlap PCR method and transformed into HMS174 cells. An aliquot mid-log phase cultures of the transformants were shocked at 48 °C and plated on LB plate (containing chloramphenicol). Afterwards, gene mutants were screened by using colony PCR and DNA sequencing. After the mutants were obtained, the phenotypes of ΔfliC and ΔdctA gene mutants were characterized by using agar puncture and carbon metabolism experiments. Colony PCR and sequencing results show that TeI3c/4c intron was inserted in the designed sites of fliC and dctA genes. The gene-targeting efficiency of Thermotargetron system was 100%. Phenotype verification experiments of the mutants demonstrated that the cell motility of all ΔfliC mutants was damaged and the malate assimilation ability of ΔdctA mutant was deprived comparing to wild-type HMS174 strain. In our study, a temperature-inducible and high-efficiency gene inactivation system was established for mesophilic bacteria. This system could achieve high efficiency and precise gene inactivation by modulation of the incubation duration of the transformants at 48 °C.

Moschus chrysogaster (sifanicus) viral hemorrhagic disease (McVHD) is an acute and highly lethal infectious disease caused by Moschus chrysogaster hemorrhagic disease virus (McHDV) whose genome sequence is highly homologous with rabbit hemorrhagic disease virus. To screen the protective antigen of McHDV and set the basis for study of McVHD vaccine, the antigen epitope of major structural protein VP60 of McHDV was analyzed, and the specific primers were designed to obtain three amplified DNA sequences encoding the main antigen epitope of VP60 from McHDV by using RT-PCR. Then the three DNA fragments were sequenced and cloned to prokaryotic expression vector with pET-28a(+) by using overlap extension PCR, and finally the prokaryotic expression plasmid pET-truncated-VP60 was constructed. Subsequently, the pET-truncated-VP60 was transformed into Escherichia coli BL21(DE3), and the recombinant proteins were expressed by IPTG induction. Finally, the expressed protein was purified and applied to immunize that without immunizing with RHD vaccine, then the antiserum titers were evaluated by the hemagglutination inhibition test, and the immune-protective efficacy of the recombinant proteins was observed and analyzed through animal challenge test. The results showed that the multi-epitope DNA fragments of VP60 of McHDV was successfully expressed in the form of inclusion bodies in E. coli, and the relative molecular weight of recombinant proteins is about 45 kDa. After immunized with the recombinant proteins, 100% of New Zealand white rabbits were resistant to attack of McHDV, which indicates efficient immune-protective efficacy of chosen epitope recombinant protein. The study laid a foundation for the development of the new subunit vaccines of McVHD.

Objective To discuss the correlation between family history of malignant neoplasms (MN-FH) and the clinicopathological features of patients with non-small cell lung cancer (NSCLC). Methods The clinicopathological data of 326 patients with NSCLC in Yangzhou University Affiliated Hospital from June 2016 to June 2018 were analyzed retrospectively. The patients were divided into two groups based on with or without MN-FH, and the clinicopathological features of the two groups were analyzed by χ 2 test. Results Of the 326 patients with NSCLC, 41 (12.6％) were in the MN-FH group and 285 (87.4％) in the MN-FH group. There were no significant differences in sex, age, smoking, location of tumors and histological classification between patients with or without MN-FH (χ 2 values were 0.031, 0.769, 0.546, 0.117, and 0.945, all P > 0.05), but in patients with MN-FH, the proportion of tumor diameter < 5 cm [65.9％ (27/41) vs. 42.5％(121/285), χ 2 = 14.173, P < 0.05], undifferentiation and low differentiation [70.7％ (29/41) vs. 53.7％(153/285), χ 2 = 4.224, P < 0.05], TNM stage Ⅲ+Ⅳ [65.9％ (27/41) vs. 46.7％ (133/285), χ 2 = 5.280, P <0.05], lymph node metastasis [78.0％ (32/41) vs. 60.0％ (171/285), χ 2 = 4.970, P < 0.05], distant metastasis [75.6％ (31/41) vs. 53.3％ (152/285), χ 2 = 7.224, P < 0.05], high degree of malignancy [70.7％ (29/41) vs. 51.6％ (147/285), χ 2 = 5.293, P < 0.05] and combination of other tumors [29.3％ (12/41) vs. 7.7％ (22/285),χ 2 = 17.817, P < 0.05] were significantly higher than those in patients without MN-FH. Conclusions NSCLC patients with MN-FH have a higher degree of malignancy. For people with MN-FH, physical examination is very important.

Objective: To investigate the effect of ultrasound transdermal delivery in children with swelling and pain after supracondylar fracture of the humerus. Methods: Totally 103 cases of humeral supracondylar fractures from January to June 2017 were divided into control group (50 cases) and experimental group (53 cases) by random number table method. The control group was treated with conventional detumescence and intravenous drug therapy,while the experimental group was given the local target ultrasonic percutaneous delivery in accordance with the doctor′s advice on the basis of routine treatment. By comparing the two groups in terms of postoperative pain, swelling relief, starting time for getting out of bed, hospitalization time, and parental satisfaction of children, we investigated the effect of ultrasonic ultrasound transdermal delivery on children with swelling and analgesia after supracondylar fracture of the humerus. Results: Postoperative pain relief and swelling relief were significantly reduced in the experimental group compared with the control group, and the difference between the two groups was statistically significant (P<0.05).The time of starting to get out of bed activity and hospitalization were respectively (8.20 ± 0.25) hours and (5.12 ± 0.27) days, and the control group was (23.80 ± 0.15) hours, (7.35 ± 0.38) days, respectively.The comparison was statistically significant (t=24.241, 7.654, all P<0.05). The parents' satisfaction in the experimental group was very satisfied, relatively satisfied, basically satisfied and unsatisfied, respectively 88.68% (47/53), 7.55% (4/53), 3.77% (2/53), 0, the control group was 64.00% (32/50), 26.00% (13/50), 10.00% (5/50), 0(0/50), the difference between the two groups was statistically significant (χ²=5.294 2, P<0.05). Conclusions The accuracy of ultrasonic percutaneous local target placement for the treatment of supracondylar fractures of the humerus in children is effective in the intervention of pain and swelling, and the comfort is better. It is a new choice for the treatment of limb fractures with pain in children.

Objective To study the expression of ultra-conserved gene sequence uc.454 in non-small cell lung cancer (NSCLC) tissues and its clinical significance.Methods Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of uc.454 mRNA in 130 cases of NSCLC and adjacent normal tissues from Affiliated Hospital of Yangzhou University between January 2014 and December 2017,and its association with clinicopathological factors and prognosis of NSCLC patients was analyzed.Results The expression level of uc.454 mRNA in NSCLC tissues was lower than that in adjacent normal tissues (2.98 ±0.96 vs.13.95± 2.23,t =98.441,P ＜ 0.05),and its expression level was associated with tumor infiltration depth (x2 =7.823,P =0.040),TNM stage (x2 =20.719,P ＜ 0.01) and lymph node metastasis (x2 =24.142,P ＜ 0.01).The survival time of patients with low expression of uc.454 mRNA was shorter than that of patients with normal or high expression (26.9 months vs.43.7 months),and the difference was statistically significant (P ＜ 0.05).Conclusion The low expression of ultra-conserved gene sequence uc.454 mRNA is related to the occurrence,development and prognosis of NSCLC.