ObjectiveTo explore the therapeutic effect and mechanism of Huangqi Xiayuxue decoction against hepatocellular carcinoma based on network pharmacology， animal experiments， and cell experiments. MethodsThe active ingredients and corresponding targets of Huangqi Xiayuxue decoction were screened via databases including the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine （BATMAN-TCM）. Hepatocellular carcinoma-related targets were retrieved from the GeneCards database. The common targets shared by Huangqi Xiayuxue decoction and hepatocellular carcinoma were subjected to Gene Ontology （GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses. Huangqi Xiayuxue decoction-containing plasma was prepared. The cell counting kit-8 （CCK-8） assay was adopted to observe the effects of different volume fractions （5%， 10%， 15%， and 20%） of Huangqi Xiayuxue decoction-containing plasma on the proliferation of HepG2， MHCC-97H， Hepa1-6， and Huh-7 cells， and the concentration for subsequent experiments was determined. Flow cytometry was adopted to detect the effects of Huangqi Xiayuxue decoction-containing plasma （5%， 10%， 15%， and 20%） on the apoptosis of MHCC-97H cells. A mouse model of subcutaneous xenograft hepatocellular carcinoma was established. The model mice were assigned into low-， medium-， and high-dose （4.63， 9.25， 18.5 g·kg^-1） Huangqi Xiayuxue decoction and sorafenib （20 mg·kg^-1） groups. Body mass and tumor volume of mice were dynamically monitored during the intervention period. After 3 weeks of intervention， hematoxylin-eosin （HE） staining was performed to observe tumor morphology. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） assay was employed to detect cell apoptosis. Immunohistochemistry （IHC） was adopted to examine the expression of cysteinyl aspartate-specific proteinase-3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the tumor tissue. Real-time PCR and Western blot were employed to verify the changes in apoptosis-related factors and the phosphatidylinositol 3-kinase/protein kinase B/forkhead box O1 （PI3K/Akt/FoxO1） signaling pathway. ResultsA total of 220 common targets shared by Huangqi Xiayuxue decoction and hepatocellular carcinoma were screened out by network pharmacology， among which tumor protein p53 （TP53）， Akt1， and signal transducer and activator of transcription 3 （STAT3） were the core targets. Pathway enrichment analysis indicated that the common targets were involved in cell apoptosis and proliferation， as well as the PI3K/Akt and FoxO signaling pathways. Cell experiments showed that compared with the blank plasma group， all volume fractions of Huangqi Xiayuxue decoction-containing plasma inhibited the proliferation of HepG2， MHCC-97H， Hepa1-6， and Huh-7 cells （P<0.01）. The proportion of early apoptotic cells in MHCC-97H cells was increased after treatment with different volume fractions of Huangqi Xiayuxue decoction-containing plasma （P<0.01）. Animal experiments revealed that Huangqi Xiayuxue decoction inhibited the growth of tumor volume， promoted tumor cell apoptosis in the high-dose group， up-regulated the protein and mRNA levels of Bax and Caspase-3， down-regulated the protein and mRNA levels of Bcl-2， and reduced the phosphorylation level of the PI3K/Akt/FoxO1 signaling pathway （P<0.05， P<0.01）. ConclusionHuangqi Xiayuxue decoction exerts anti-hepatocellular carcinoma effects by inhibiting the activation of the PI3K/Akt/FoxO1 signaling pathway and promoting hepatocellular carcinoma cell apoptosis.

ObjectiveTo explore the therapeutic effect and mechanism of Huangqi Xiayuxue decoction against hepatocellular carcinoma based on network pharmacology， animal experiments， and cell experiments. MethodsThe active ingredients and corresponding targets of Huangqi Xiayuxue decoction were screened via databases including the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine （BATMAN-TCM）. Hepatocellular carcinoma-related targets were retrieved from the GeneCards database. The common targets shared by Huangqi Xiayuxue decoction and hepatocellular carcinoma were subjected to Gene Ontology （GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses. Huangqi Xiayuxue decoction-containing plasma was prepared. The cell counting kit-8 （CCK-8） assay was adopted to observe the effects of different volume fractions （5%， 10%， 15%， and 20%） of Huangqi Xiayuxue decoction-containing plasma on the proliferation of HepG2， MHCC-97H， Hepa1-6， and Huh-7 cells， and the concentration for subsequent experiments was determined. Flow cytometry was adopted to detect the effects of Huangqi Xiayuxue decoction-containing plasma （5%， 10%， 15%， and 20%） on the apoptosis of MHCC-97H cells. A mouse model of subcutaneous xenograft hepatocellular carcinoma was established. The model mice were assigned into low-， medium-， and high-dose （4.63， 9.25， 18.5 g·kg^-1） Huangqi Xiayuxue decoction and sorafenib （20 mg·kg^-1） groups. Body mass and tumor volume of mice were dynamically monitored during the intervention period. After 3 weeks of intervention， hematoxylin-eosin （HE） staining was performed to observe tumor morphology. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） assay was employed to detect cell apoptosis. Immunohistochemistry （IHC） was adopted to examine the expression of cysteinyl aspartate-specific proteinase-3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the tumor tissue. Real-time PCR and Western blot were employed to verify the changes in apoptosis-related factors and the phosphatidylinositol 3-kinase/protein kinase B/forkhead box O1 （PI3K/Akt/FoxO1） signaling pathway. ResultsA total of 220 common targets shared by Huangqi Xiayuxue decoction and hepatocellular carcinoma were screened out by network pharmacology， among which tumor protein p53 （TP53）， Akt1， and signal transducer and activator of transcription 3 （STAT3） were the core targets. Pathway enrichment analysis indicated that the common targets were involved in cell apoptosis and proliferation， as well as the PI3K/Akt and FoxO signaling pathways. Cell experiments showed that compared with the blank plasma group， all volume fractions of Huangqi Xiayuxue decoction-containing plasma inhibited the proliferation of HepG2， MHCC-97H， Hepa1-6， and Huh-7 cells （P<0.01）. The proportion of early apoptotic cells in MHCC-97H cells was increased after treatment with different volume fractions of Huangqi Xiayuxue decoction-containing plasma （P<0.01）. Animal experiments revealed that Huangqi Xiayuxue decoction inhibited the growth of tumor volume， promoted tumor cell apoptosis in the high-dose group， up-regulated the protein and mRNA levels of Bax and Caspase-3， down-regulated the protein and mRNA levels of Bcl-2， and reduced the phosphorylation level of the PI3K/Akt/FoxO1 signaling pathway （P<0.05， P<0.01）. ConclusionHuangqi Xiayuxue decoction exerts anti-hepatocellular carcinoma effects by inhibiting the activation of the PI3K/Akt/FoxO1 signaling pathway and promoting hepatocellular carcinoma cell apoptosis.

Objective:To quantify the associations between periconceptional maternal homocysteine (HCY) and offspring′s birth weight and risk of small for gestational age (SGA) infant.Methods:The 19 984 mother-child pairs in this prospective cohort study were recruited from the Shanghai preconception cohort; the infants were delivered from 1 st September 2016 to 11 th November 2022. A standardized questionnaire was used to collect the mothers′ demographic information, medical history, dietary supplement use, and maternal complications during pregnancy, and their serum samples were collected. Serum HCY, folate, and vitamin B 12 were measured using chemiluminescent microparticle immunoassay based on serum sample drawn at enrollment. Birth weight data were obtained from medical records. Multiple imputation methods were applied to handle missing data in key variables. Multivariable linear regression and Poisson regression models were used to analyze the relationship between maternal HCY concentration during the periconceptional period and the birth weight and SGA risk of the offspring. Results:A total of 9 452 pairs were enrolled preconceptionally and the remaining 10 532 pairs were enrolled in early pregnancy. The proportion of mothers whose pregnancy age was greater than 35 years was 9.2% (1 832/19 984), the proportion of primiparous women was 76.5% (15 283/19 984), the proportion of pre-pregnancy overweight and obesity was 14.0% (2 804/19 984), the proportion of using folic acid supplements before pregnancy was 21.4% (4 272/19 984), and the proportion of those who supplemented with folic acid during early pregnancy was 85.2% (8 976/10 532); gestational diabetes mellitus was in 6.2% (1 245/19 984), gestational hypertensive syndrome in 3.6% (711/19 984). The birth weight of the offspring was (3 297±468) g, and there were 1 962 SGA children (9.8%). The HCY concentration in the overall population in appropriate for gestational age during the periconceptional period was (7.9±3.2) μmol/L, with (8.3±3.7) μmol/L in the preconception subgroup and (7.3±2.4) μmol/L in the early pregnancy subgroup. After adjustment for the covariates of perinatal demographic information, adverse pregnancy outcomes, serum folate and vitamin B 12, increased maternal periconceptional HCY was significantly associated with lower offspring birth weight ( β=-2.30, 95% CI -4.43--0.16, P=0.035). Only the early pregnancy subgroup was significantly associated with lower offspring birth weight ( β=-7.39, 95% CI-11.50--3.21, P<0.001). No association was found between peripregnancy HCY and offspring SGA risk. However, elevated HCY in early pregnancy was associated with an increased risk of SGA in the offspring ( RR=1.05, 95% CI 1.01-1.08, P=0.002). Periconceptional vitamin B 12 was a mediator of the association between HCY and offspring birth weight, accounting for 16.5%, 41.2% and 5.4% of its total effect in the overall periconceptional population, the pre-pregnancy subgroup and the early pregnancy subgroup, respectively. Conclusions:Maternal periconceptional HCY level is associated with lower birth weight in offspring, but not with the risk of SGA. Elevated maternal HCY in early pregnancy subgroup may be associated with increased risk of SGA in offspring.

Objective: To evaluate the immunity and influencing factors of diphtheria among preschool children in Shenzhen,to provide reference for effective monitoring of diphtheria IgG antibody level in preschool children. Methods: Serum samples were collected from 296 preschool children aged 4-6 who were recruited in Shenzhen. The diphtheria antibody titer in serum was determined by enzyme linked immunosorbent assay, and the effect of different immumuzation schedule including types of vaccine and vaccination timing, on the geometric mean concentration (GMC) of diphtheria IgG antibody and antibody positive rate were analyzed. Results: The GMC of diphtheria IgG antibody was 0.71 IU/mL, and the positive conversion rate was 33.1%. There were significant differences in antibody GMC and antibody positive conversion rate of diphtheria in different age groups( F/χ 2=11.77, 27.45, P < 0.01 ). The GMC and antibody positive conversion rate showed significant differences by diphtheria antibodies, vaccine types and end dose vaccination intervals( F=49.53, 12.95，11.61, P <0.01). There were statistically significant differences in the positive conversion rate of diphtheria antibodies in children with different types of diphtheria antibodies, vaccine types of diphtheria antibodies, and diphtheria antibodies at the time interval of final vaccination (Fisher exact probability method, P <0.01). Conclusion The overall positive conversion rate of diphtheria antibody in preschool children in Shenzhen is high. Timely completion of full diphtheria vaccination can improve the antibody level and plays a better role in protecting preschool children.

Objective: To evaluate the effect of a dietary and exercise intervention on cognition and behaviors among primary school students, and to further explore the mediating role of family support, in order to provide scientific evidence for future effective intervention strategies. Methods: A dietary and exercise intervention program for childhood obesity prevention was carried out in 24 primary schools in Beijing, Changzhi and Urumqi from 2018 to 2019, and 1 392 children in grade four as well as their parents were included. Family support and children s cognition and behaviors were collected through questionnaire. To carry out diet and exercise behavior intervention on the three levels of school, family and individual in the intervention group,and children s dietary and exercise knowledge, eating habits, physical activities, as well as sedentary and screen behaviors were evaluated. The linear mixed model was used to analyze the effect of intervention on children s cognition and behaviors, and the mediation model was used to explore the role of family support. Results: The proportion of children with higher score of dietary and exercise knowledge ( OR= 2.34 , 95%CI =1.71-3.21), eating habits ( OR=2.58, 95%CI =1.75-3.82), and sedentary and screen behaviors ( OR=1.91, 95%CI =1.35-2.68) increased in the intervention group after one year intervention ( P <0.01), compared with the control group, respectively. The intervention also increased the proportion of children s family support in the intervention group compared with the control group ( OR=3.45, 95%CI =2.19-5.45), and the support from children s fathers ( OR=2.70, 95%CI =1.68-4.35), mothers ( OR=3.71, 95%CI =2.28-6.04), paternal grandmothers ( OR=1.65, 95%CI =1.00-2.70), and maternal grandmothers ( OR= 2.14, 95%CI =2.12-2.16) increased significantly ( P <0.05). The mediation analysis showed that family support played a mediating role in association between comprehensive intervention and children s eating habits as well as sedentary and screen behaviors. Conclusion The dietary and exercise intervention effectively promoted children s cognition and behaviors, and family support played an important mediating role.

Objective:To compare the advantages of high or low inferior mesenteric artery (IMA) ligation in laparoscopic anterior resection of rectum based on CT precise guidance.Methods:One hundred and twenty patients with rectal cancer who underwent laparoscopic anterior resection of rectum in Shanghai Sixth People′s Hospital South Campus from January 2016 to June 2020 were selected. Surgical protocol was selected according to the principle of voluntariness. Sixty-eight cases underwent high IMA ligation (high IMA ligation group), and 52 cases underwent low IMA ligation (low IMA ligation group). The perioperative conditions (operative time, intraoperative blood loss, postoperative anal exhaust time, abdominal drainage, hospitalization time and ileostomy), postoperative pathological data (proximal margin distance, distal margin distance, total number of dissected lymph nodes and number of positive lymph nodes) and postoperative recovery (anastomotic stoma fistula, urinary retention, sexual dysfunction and daily number of defecation) were recorded. The quality of life 6 months after surgery was evaluated by the quality of life instruments for colorectal cancer scale (QLQ-CR38). Patients were followed up for 6 to 36 months, and the local recurrence and distant metastasis were recorded.Results:The postoperative anal exhaust time in low IMA ligation group was significantly shorter than that in high IMA ligation group: (2.87 ± 1.04) d vs. (3.26 ± 1.00) d, and there was statistical difference ( P<0.05); there were no statistical differences in operative time, intraoperative blood loss, abdominal drainage, hospitalization time and rate of ileostomy between 2 groups ( P>0.05). There were no statistical difference in proximal margin distance, distal margin distance, total number of dissected lymph nodes and number of positive lymph nodes between 2 groups ( P>0.05). The incidences of anastomotic stoma fistula, urinary retention and sexual dysfunction in low IMA ligation group were significantly lower than those in high ligation group: 3.85% (2/52) vs. 13.24% (9/68), 3.85% (2/52) vs. 11.76% (8/68) and 5.77% (3/52) vs. 14.71% (10/68), the daily number of defecation was significantly less than that in high ligation group: (2.87 ± 0.98) times vs. (4.05 ± 1.56) times, and there were statistical differences ( P<0.05 or <0.01). The functional dimension, symptom dimension and aggregate score of QLQ-CR38 in low IMA ligation group were significantly higher than those in high ligation group: (15.46 ± 4.22) scores vs. (13.68 ± 3.56) scores, (51.82 ± 13.54) scores vs. (45.65 ± 12.42) scores and (67.28 ± 14.28) scores vs. (59.33 ± 12.85) scores, and there were statistical differences ( P<0.05 or <0.01). There were no statistical difference in the rates of local recurrence and distant metastasis between 2 groups ( P>0.05). Conclusions:The application of low IMA ligation based on CT precise guidance in laparoscopic anterior resection of rectum can obtain the same surgical effect as the high IMA ligation. It can promote the recovery of gastrointestinal function after surgery, reduce the risk of complications, have less effect on the defecation function, which can improve the life quality after the surgery.

Objective To investigate the therapeutic effect of anteromedial ankle approach for talus fracture.Methods From January 2012 to October 2018,the data of 28 cases of talus fracture treated by anteromedial ankle approach were retrospectively analyzed,including 26 males and 2 females,aged 24-61 years with an average of 38.6 years.All 25 cases of closed injuries and 3 cases of open injuries were fresh injuries.There were 16 cases of traffic injuries,8 falling injuries,2 heavy object injuries and 2 sprains.Among the 28 patients,17 were talus neck fractures,and according to Hawkins talus neck fracture classification,there were 1 case of type Ⅰ,12 type Ⅱ,and 4 type Ⅲ.There were 11 cases of talus fracture,which were classified according to Sneppen talus fracture types:2 cases of type Ⅰ,8 cases of type Ⅱ and 1 case of type Ⅴ.Among them,2 cases of talus neck fracture complicated with T12 vertebral body fractures and 2 cases complicated with calcaneal fractures.The time from injury to operation for 3 patients with open fracture was 3-6 hours.25 cases of closed fracture injuries took 2-10 hours to operate,with an average time of 5.6 hours.All patients were fixed with cannulated screws through the anteromedial ankle approach.The weight-bearing time and strength of lower limbs were determined according to fracture healing after surgery.The ankle function was evaluated by American Orthopaedic Foot and Ankle Society (AOFAS) Score.Results All 28 patients were followed up for 23 to 82 months,with an average of 37.5 months.All fractures healed.The healing time of talus neck fracture was 3.5-8 months,with an average of 5 months.The healing time of talus fracture was 3-7 months,averaging 4.5 months.At the latest follow-up,AOFAS score ranged from 68 to 100,with an average of 82.5 points,including excellent in 15 cases,good in 8 cases,fair in 4 cases and poor in 1 case,with the excellent and good rate of 82.1％ (23/28).There were no signs of fracture displacement,malunion,loosening and prolapse of internal fixation,fracture and ischemic necrosis of talus after operation.Traumatic arthritis occurred in 9 patients,with an incidence rate of 32.1％ (9/28).Tibial joint was involved in 5 cases,talus joint in 3 cases,tibialis in 1 case and talus joint in 1 case.Among them,traumatic arthritis occurred in 2 cases of open injury.Nine patients (32.1％) suffered from traumatic arthritis 6-10 months after operation,with an average of 8 months.According to conservative treatments such as weight control,ankle immobilization,physiotherapy,anti-inflammatory and analgesic,nutrition of articular cartilage drugs,joint traction and plasty were given.The patients' conditions improved to different degrees and could tolerate walking and normal life and work.Conclusion The anteromedial ankle approach has the advantages of minimal vessel injuries,clear exposure for direct reduction,low rate of necrosis of talus and satisfactory curative effect.

Progression of tumor is a complex process with multiple steps and multiple factors.MicroRNA-296 (miR-296) plays an important role in tumor progression,involved in the proliferation,differentiation,angiogenesis,invasion,metastasis,apoptosis and drug resistance of the tumor cells.

BACKGROUND:Screening of fare blood types has been successively implemented and completed in Europe, America and Japan, but there is a large gap in China. Previous studies have mainly focused on the southern Han populations, and little is reported on PCR-SSP systematic analysis of gene frequencies of rare blood groups in Xinjiang Uygur populations. OBJECTIVE:To investigate the gene frequency distribution of RBC MNS, Duffy, Kel, Dombrock, Diego, Kidd, Scianna, Colton and Lutheran blood groups from Xinjiang Uygur populations, thereby providing a strategic support for human population genetics and clinical blood deployment. METHODS:PCR-SSP method was used to make genotyping and statistical analysis in 158 Xinjiang Uygur persons from nine rare blood groups. RESULTS AND CONCLUSION: Gene frequencies of these nine rare blood groups were M=0.579 1, N=0.420 9, S=0.174 3, s=0.800 9, Fya=0.699 4, Fyb=0.300 6, K1=0.015 8, K2=0.984 2, Doa=0.234 2, Dob=0.765 8, Dia=0.047 4, Dib=0.952 6, JKa=0.541 2, JKb=0.452 6, Sc1=1.000, Sc2=0, Coa=0.994, Cob=0.005 9, Lua=0, Lub=1.000, Aua=0.810 2, Aub=0.189 9. Results from chi-square test showed that the observed value and expected value of genotypes were in line with the law of Hardy-Weinberg equilibrium (P > 0.05), and in the MNS blood group of Xinjiang Uygur population, it was rarely found that S-s- frequency was 0.025 3 in four cases and Jka-b- frequency was 0.006 3 in one case. This study demonstrates that the frequency distribution of MNS, Duffy, Dombrock and Diego blood groups in the Xinjiang Uygur population, with its own unique frequency distribution characteristics, is different from that in other ethnic populations; the gene distribution of Kel, Kidd and Colton blood groups shows either similarity or difference between the Xinjiang Uygur population and reported Tibet and Han populations; Scianna and Lutheran blood groups show a monomorphic distribution in the Xinjiang Uygur population, which is similar to that in the Tibet and Han populations. These findings provide the basic data for exploring the origin and evolution, ethnic hematology and construction of rare blood database of the Xinjiang Uygur population. Cite this article:Lin GY, Du XL, Shan JJ, Zhang YN, Zhang YQ, Zhang YZ.Molecular genetic analysis of genes from MNS, Duffy and Kel blood groups in the China Xinjiang Uygur population. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):123-127.

OBJECTIVETo identify mutations of dystrophin gene in a Chinese pedigree affected with Duchenne muscular dystrophy (DMD).

METHODSClinical data from the pedigree was collected. Subsequently, polymerase chain reaction and DNA sequencing analysis were applied to detect the potential mutations. Restriction enzyme digestion was carried out to determine whether the mutation was present in 118 healthy controls. Clustal software was applied for analyzing the conservation of altered amino acids.

RESULTSDNA sequencing analysis has identified a heterozygous missense mutation c.7578G>C (p.Gln2526His) mutation in exon 52 of the dystrophin gene in the proband and his mother. The same mutation was absent in the 118 healthy controls. Restriction enzyme digestion has confirmed above result. Clustal analysis indicated that the altered amino acid is highly conserved in mammals.

CONCLUSIONThe results revealed a novel missense mutation (c.7578G>C) of the dystrophin gene in DMD patients.

Adolescent ; Adult ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Child ; Child, Preschool ; China ; Dystrophin ; genetics ; Exons ; Female ; Humans ; Male ; Molecular Sequence Data ; Muscular Dystrophy, Duchenne ; genetics ; Mutation, Missense ; Pedigree ; Young Adult