OBJECTIVE To optimize the extraction process of flavonoids from Potentilla discolor .METHODS Deep eutectic solvents （DESs） were used as the extraction solvent to prepare test sample solutions. The contents of rutin， quercitrin， and kaempferol were determined by ultra-performance liquid chromatography. The analytic hierarchy process combined with the criteria importance through intercriteria correlation weighting method was employed to assign comprehensive weights to the contents of the three flavonoids， and calculate the comprehensive score. The optimal DESs were screened based on the comprehensive score.Mean while， using the comprehensive score as the response variable and the liquid-to-solid ratio， ultrasonic time， and urltrasonic temperature as the influencing factors， the extraction technology of flavonoids from P.discolor was optimized by Box-Behnken response surface methodology and subsequently validated.RESULTS The comprehensive weight coefficients of rutin， quercitrin， and kaempferol were 0.646 7， 0.207 5， and 0.145 8， respectively. The optimal DESs were choline chloride-lactic acid （molar ratio 1∶4） with a water content of 40%. The optimal extraction conditions were determined as follows： liquid-to-solid ratio of 25∶1， ultrasonic time of 40 min， and ultrasonic temperature of 50 ℃. The validation results showed that the average comprehensive score of three batches of samples was 99.28， with an RSD of 0.54% （ n ＝3）.CONCLUSIONS The optimal extraction technology of flavonoids from P. discolor based on DESs obtained in this study was stable and feasible.

Non-alcoholic fatty liver disease(NAFLD)is a chronic liver disease caused by excessive lipid accumulation in the liver.Its incidence rate is increasing year by year and has become an increasingly serious public healthy problem.The pathogenesis of NAFLD is complex and has not been fully clarified at present.It is mainly related to multiple factors such as genetics,metabolism,intestinal flora and immune response.In order to explore the medication rules and mechanism of action of traditional Chinese medicine(TCM)in the treatment of NAFLD,and to provide references for the treatment of NAFLD with TCM and the research and development of new drugs,this article summarizes the TCM pathogenesis of NAFLD(such as"phlegm and blood stasis interlacing","liver depression and spleen deficiency",etc.)and modern etiology and pathogenesis(such as insulin resistance,lipid disorder,mitochondrial dysfunction,oxidative stress,etc.).The clinical research and experimental data at home and abroad in recent years were integrated to analyze the pathological process of NAFLD intervention by TCM through multiple targets,including improving insulin resistance and lipid metabolism disorders,inhibiting oxidative stress and mitochondrial dysfunction,etc.TCM has shown unique advantages in the prevention and treatment of NAFLD.However,the depth of its mechanism analysis and the level of clinical research still need to be improved.In the future,it is necessary to deepen the mechanism research by combining multi-omics technology to accelerate the modernization development of TCM.

Non-alcoholic fatty liver disease(NAFLD)is a chronic liver disease caused by excessive lipid accumulation in the liver.Its incidence rate is increasing year by year and has become an increasingly serious public healthy problem.The pathogenesis of NAFLD is complex and has not been fully clarified at present.It is mainly related to multiple factors such as genetics,metabolism,intestinal flora and immune response.In order to explore the medication rules and mechanism of action of traditional Chinese medicine(TCM)in the treatment of NAFLD,and to provide references for the treatment of NAFLD with TCM and the research and development of new drugs,this article summarizes the TCM pathogenesis of NAFLD(such as"phlegm and blood stasis interlacing","liver depression and spleen deficiency",etc.)and modern etiology and pathogenesis(such as insulin resistance,lipid disorder,mitochondrial dysfunction,oxidative stress,etc.).The clinical research and experimental data at home and abroad in recent years were integrated to analyze the pathological process of NAFLD intervention by TCM through multiple targets,including improving insulin resistance and lipid metabolism disorders,inhibiting oxidative stress and mitochondrial dysfunction,etc.TCM has shown unique advantages in the prevention and treatment of NAFLD.However,the depth of its mechanism analysis and the level of clinical research still need to be improved.In the future,it is necessary to deepen the mechanism research by combining multi-omics technology to accelerate the modernization development of TCM.

Objective:To understand the current status of anemia, iron deficiency, and iron-deficiency anemia among preschool children in China.Methods:A cross-sectional study was conducted with a multi-stage stratified sampling method to select 150 streets or townships from 10 Chinese provinces, autonomous regions, or municipalities (East: Jiangsu, Zhejiang, Shandong, and Hainan; Central: Henan; West: Chongqing, Shaanxi, Guizhou, and Xinjiang; Northeast: Liaoning). From May 2022 to April 2023, a total of 21 470 children, including community-based children aged 0.5 to<3.0 years receiving child health care and kindergarten-based children aged 3.0 to<7.0 years, were surveyed. They were divided into 3 age groups: infants (0.5 to<1.0 year), toddlers (1.0 to<3.0 years), and preschoolers (3.0 to<7.0 years). Basic information such as sex and date of birth of the children was collected, and peripheral blood samples were obtained for routine blood tests and serum ferritin measurement. The prevalence rates of anemia, iron deficiency, and iron-deficiency anemia were analyzed, and the prevalence rate differences were compared among different ages, sex, urban and rural areas, and regions using the chi-square test.Results:A total of 21 460 valid responses were collected, including 10 780 boys (50.2%). The number of infants, toddlers, and preschoolers were 2 645 (12.3%), 6 244 (29.1%), and 12 571 (58.6%), respectively. The hemoglobin level was (126.7±14.8) g/L, and the serum ferritin level was 32.3 (18.5, 50.1) μg/L. The overall rates of anemia, iron deficiency, and iron-deficiency anemia were 10.4% (2 230/21 460), 28.3% (6 070/21 460), and 3.9% (845/21 460), respectively. The prevalence rate of anemia was higher for boys than for girls (10.9% (1 173/10 780) vs. 9.9% (1 057/10 680), χ2=5.58, P=0.018), with statistically significant differences in the rates for infants, toddlers and preschoolers (18.0% (475/2 645), 10.6% (662/6 244), and 8.7% (1 093/12 571), respectively, χ2=201.81, P<0.01), and the rate was significantly higher for children in rural than that in urban area (11.8% (1 516/12 883) vs. 8.3% (714/8 577), χ2=65.54, P<0.01), with statistically significant differences in the rates by region ( χ2=126.60, P<0.01), with the highest rate of 15.8% (343/2 173) for children in Central region, and the lowest rate of 5.3% (108/2 053) in Northeastern region. The prevalence rates of iron deficiency were 33.8% (895/2 645), 32.2% (2 011/6 244), and 25.2% (3 164/12 571) in infants, toddlers, and preschoolers, respectively, and 30.0% (3 229/10 780) in boys vs. 26.6% (2 841/10 680) in girls, 21.7% (1 913/8 821), 40.0% (870/2 173), 27.1% (2 283/8 413), 48.9% (1 004/2 053) in Eastern, Central, Western, and Northeastern regions, respectively, and each between-group showed a significant statistical difference ( χ2=147.71, 29.73, 773.02, all P<0.01). The prevalence rate of iron-deficiency anemia showed a significant statistical difference between urban and rural areas, 2.9% (251/8 577) vs. 4.6% (594/12 883) ( χ2=38.62, P<0.01), while the difference in iron deficiency prevalence was not significant ( χ2=0.51, P=0.476). Conclusions:There has been a notable improvement in iron deficiency and iron-deficiency anemia among preschool children in China, but the situation remains concerning. Particular attention should be paid to the prevention and control of iron deficiency and iron-deficiency anemia, especially among infants and children in the Central, Western, and Northeastern regions of China.

Objective This study aimed to explore the heterogeneity and gene ontology of Wnt1-Cre-marked and Pax2-Cre-marked first branchial arch cranial neural crest cells(CNCs)in mice.Methods The embryos of Wnt1-Cre;R26RmTmG and Pax2-Cre;R26RmTmG at embryonic day(E)8.0-E9.25 were collected for histological observation.We per-formed immunostaining to compare green fluorescent pro-tein(GFP)-positive CNCs in Pax2-Cre;R26RAi9 and Wnt1-Cre;R26RAi9 mice at E15.5.Single-cell RNA sequencing(scRNA-seq)was used to analyze the first branchial arch GFP-positive CNCs from Wnt1-Cre;R26RmTmG and Pax2-cre;R26RmTmGmice at E10.5.Real time fluorescence quantitative polymerase chain reaction(q-PCR)was performed to val-idate the differential genes.Results Wnt1-Cre-marked and Pax2-Cre-marked CNCs migrated from the neural plateto first and second branchial arches and to the first branchial arch,respectively,at E8.0.Although Wnt1-Cre-marked and Pax2-Cre-marked CNCs were found mostly in cranial-facial tissues,the former had higher expression in palate and tongue.The results of scRNA-seq showed that Pax2-Cre-marked CNCs specifically contributed to osteoblast differentia-tion and ossification,while Wnt1-Cre-marked CNCs participated in limb development,cell migration,and ossification.The q-PCR data also confirmed the results of gene ontology analysis.Conclusion Pax2-Cre mice are perfect experimen-tal animal models for research on first branchial arch CNCs and derivatives in osteoblast differentiation and ossification.

Objective:To explore the relationship between dietary pattern and sexual function in people of childbearing age.Methods:This study adopted a cross-sectional method and included all women and men who visited Reproduction Center of Maanshan Maternal and Child Health Hospital, Reproductive Medicine Center of the 901th Hospital of the Joint Logistics Support Force of People's Liberation Army and Reproduction Center of Anhui Wanbei Coal Power Group General Hospital from December 2020 to March 2022. The Female Sexual Function Index (FSFI) and International Index of Erectile Function-15 (IIEF-15) were used to assess sexual function. Factor analysis was used to establish dietary patterns, and multivariate logistic regression was used to analyze the association between dietary patterns and sexual function.Results:A total of 1 290 females and 1 331 males were included in the study, including 1 031 females and 899 males with sexual dysfunction. There were 289 women and 272 men with balanced dietary pattern, 473 women and 395 men with traditional dietary pattern, 272 women and 324 men with processed dietary pattern, and 256 women and 340 men with beverage dietary pattern. After adjusting for confounding factors such as residence and age, it was found that balanced dietary pattern was negatively correated with female sexual desire disorder ( OR=0.904, 95% CI: 0.820-0.995, P=0.039), sexual arousal disorder ( OR=0.840, 95% CI: 0.759-0.929, P=0.001), vaginal lubrication disorder ( OR=0.833, 95% CI: 0.710-0.979, P=0.026), orgasmic disorder ( OR=0.764, 95% CI: 0.680-0.858, P<0.001), low sexual satisfaction ( OR=0.887, 95% CI: 0.796-0.987, P=0.028), sexual dysfunction ( OR=0.805, 95% CI: 0.714-0.907, P<0.001), and male orgasmic disorder ( OR=0.859, 95% CI: 0.763-0.967, P=0.012). The traditional dietary pattern was negatively correlated with female sexual desire disorder ( OR=0.879, 95% CI: 0.786-0.983, P=0.024), sexual arousal disorder ( OR=0.884, 95% CI: 0.784-0.995, P=0.042), male erectile disorder ( OR=0.736, 95% CI: 0.634-0.855, P<0.001), sexual desire disorder ( OR=0.753, 95% CI: 0.648-0.876, P<0.001), and sexual dysfunction ( OR=0.769, 95% CI: 0.653-0.907, P=0.020). The processed dietary pattern was positively correlated with erectile disorder ( OR=1.162, 95% CI: 1.049-1.287, P=0.004), orgasmic dysfunction ( OR=1.207, 95% CI: 1.091-1.337, P<0.001), sexual desire disorder ( OR=1.199, 95% CI: 1.081-1.330, P=0.001) and sexual dysfunction ( OR=1.134, 95% CI: 1.020-1.261, P=0.002). Beverage dietary pattern was not associated with sexual dysfunction in men and women (all P>0.05). Conclusion:Balanced, traditional dietary patterns were related to the reduce risk of sexual dysfunction in both women and men of childbearing age, while processed dietary patterns were related to the increased risk of sexual dysfunction in men of childbearing age.

Objective:To explore the relationship between dietary pattern and sexual function in people of childbearing age.Methods:This study adopted a cross-sectional method and included all women and men who visited Reproduction Center of Maanshan Maternal and Child Health Hospital, Reproductive Medicine Center of the 901th Hospital of the Joint Logistics Support Force of People's Liberation Army and Reproduction Center of Anhui Wanbei Coal Power Group General Hospital from December 2020 to March 2022. The Female Sexual Function Index (FSFI) and International Index of Erectile Function-15 (IIEF-15) were used to assess sexual function. Factor analysis was used to establish dietary patterns, and multivariate logistic regression was used to analyze the association between dietary patterns and sexual function.Results:A total of 1 290 females and 1 331 males were included in the study, including 1 031 females and 899 males with sexual dysfunction. There were 289 women and 272 men with balanced dietary pattern, 473 women and 395 men with traditional dietary pattern, 272 women and 324 men with processed dietary pattern, and 256 women and 340 men with beverage dietary pattern. After adjusting for confounding factors such as residence and age, it was found that balanced dietary pattern was negatively correated with female sexual desire disorder ( OR=0.904, 95% CI: 0.820-0.995, P=0.039), sexual arousal disorder ( OR=0.840, 95% CI: 0.759-0.929, P=0.001), vaginal lubrication disorder ( OR=0.833, 95% CI: 0.710-0.979, P=0.026), orgasmic disorder ( OR=0.764, 95% CI: 0.680-0.858, P<0.001), low sexual satisfaction ( OR=0.887, 95% CI: 0.796-0.987, P=0.028), sexual dysfunction ( OR=0.805, 95% CI: 0.714-0.907, P<0.001), and male orgasmic disorder ( OR=0.859, 95% CI: 0.763-0.967, P=0.012). The traditional dietary pattern was negatively correlated with female sexual desire disorder ( OR=0.879, 95% CI: 0.786-0.983, P=0.024), sexual arousal disorder ( OR=0.884, 95% CI: 0.784-0.995, P=0.042), male erectile disorder ( OR=0.736, 95% CI: 0.634-0.855, P<0.001), sexual desire disorder ( OR=0.753, 95% CI: 0.648-0.876, P<0.001), and sexual dysfunction ( OR=0.769, 95% CI: 0.653-0.907, P=0.020). The processed dietary pattern was positively correlated with erectile disorder ( OR=1.162, 95% CI: 1.049-1.287, P=0.004), orgasmic dysfunction ( OR=1.207, 95% CI: 1.091-1.337, P<0.001), sexual desire disorder ( OR=1.199, 95% CI: 1.081-1.330, P=0.001) and sexual dysfunction ( OR=1.134, 95% CI: 1.020-1.261, P=0.002). Beverage dietary pattern was not associated with sexual dysfunction in men and women (all P>0.05). Conclusion:Balanced, traditional dietary patterns were related to the reduce risk of sexual dysfunction in both women and men of childbearing age, while processed dietary patterns were related to the increased risk of sexual dysfunction in men of childbearing age.

Objective:To explore the ability of sequential immunization regimen inducing neutralizing antibodies against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) Wuhan-Hu1 and Omicron variants in mice.Methods:Groups of 6-8-week-old BALB/c mice were primed with two doses of Wuhan-Hu-1 inactivated vaccine, and then boosted with Omicron or Wuhan-Hu-1 inactivated vaccine, respectively. Binding antibodies were tested by enzyme linked immunosorbent assay; and neutralizing antibodies against Wuhan-Hu1 and Omicron variants were analyzed by vesicular stomatitis virus pseudovirus assay system; SARS-CoV-2 specific cellular immune responses were quantified by enzyme-linked immunosorbent spot assay.Results:IgG antibodies against Wuhan-Hu1, Delta and Omicron RBD were enhanced after the second dose of Wuhan-Hu1 inactivated vaccine. Compared with Wuhan-Hu1 inactivated vaccine, the group boosted with Omicron inactivated vaccine improved Wu-RBD and Omic-RBD specific IgG antibodies 1.41 and 1.26 times, and serum neutralizing antibodies against BA.1, BA.2, BA.4/5 and BF.7 were elevated 4.5, 3.4, 12.1 and 6.5 folds, respectively, by sequential immunization. After booster immunization with inactivated Wuhan-Hu1 or Omicron vaccines, Wu-RBD IgA titer was significantly higher than that of one dose inactivated Wuhan-Hu1 vaccine ( P=0.005 7, P=0.006 1). Conclusions:Neutralizing antibodies against Omicron variants were enhanced by sequential immunization with Omicron inactivated vaccine. Specific IgA was significantly enhanced after the third dose of inactivated vaccine.

Objective:To construct standard strains representing the main epidemic clades of HIV-1 in China, amplify the virus strains, and establish a seed lot.Methods:Six isolates of HIV-1 virus were identified and analyzed in genotype and phenotype, according to " interpretation for the social organization of the Standard strains of pathogenic microorganism- technical specifications for establishment of HIV strains". The isolates were amplified and cultivated to generate the secondary generation primary seed lot and the third generation working seed lot as frozen storage in liquid nitrogen. Results:Six HIV-1 standard strains were obtained, of which 3 strains are CRF_ 07BC (NRPC2.4.9003, NRPC2.4.9005, NRPC2.4.9006), 1 strain is CRF_ 01AE (NRPC2.4.9001), 1 strain is CRF_ 08BC (NRPC2.4.9002), and 1 strain is URF (NRPC2.4.9004). Phenotypic detection showed that all six strains are CCR5 tropics and Non syncytia inducing virus. TCID 50 were all greater than 1 × 10 5/ml, and concentrations of p24 antigen were all higher than 2 ng/ml. A primary seed lot with no less than 20 vials per strain and a working seed lot with no less than 50 vials per strain were constructed. The standard virus strains were used in evaluating antiviral drugs PEG2kC34, PEG5kC34, LP-19, and neutralizing antibody LSEVh LS-F. Conclusions:Six standard strains of HIV-1 virus covering the three main epidemic subtypes of HIV-1 in China have been obtained, and a storage of HIV-1 standard strain was constructed. It meets the need of the preservation of HIV-1 standard strains in China and provides support for drug and vaccine evaluation.

Objective:To investigate the effects of oxycodone multimodal analgesia on quality of awakening and hemodynamics in patients undergoing laparoscopic gynaecological surgery.Methods:Ninety patients who underwent laparoscopic ovarian cyst removal in Weihai Central Hospital from September 2018 to March 2019 were included in this study. They were randomly divided into observation and control groups, with 45 patients in each group. Both groups were given intravenous flurbiprofen axetil (1 mg/kg) for preemptive analgesia. Combined intravenous and inhalation anesthesia was used. The depth of anesthesia was monitored. The observation group was intravenously given 0.10 mg/kg oxycodone and the control group was intravenously given 5 μg sufentanil. Quality of awakening, hemodynamic indexes, postoperative pain score, and incidences of nausea and vomiting were compared between the two groups.Results:Cough score in the observation group was significantly lower than that in the control group [(1.1 ± 0.4) points vs. (1.7 ± 0.7) points, t = -4.99, P < 0.05]. Ramsay Sedation Scale score in the observation group was significantly higher than that in the control group [(3.6 ± 1.0) points vs. (2.8 ± 0.8) points, t = 7.44, P < 0.05]. At 0 (T 1) and 5 minutes (T 2) after extubation, systolic blood pressure in the observation group was (117.7 ± 18.2) mmHg and (118.1 ± 16.2) mmHg, respectively, which were significantly lower than (134.2 ± 16.2) mmHg and (134.5 ± 15.2) mmHg in the control group ( t =-4.54, -4.95, both P < 0.05). There were no significant differences in the incidences of nausea and vomiting between the two groups (both P > 0.05). At 6 and 12 hours after surgery, visual analogue scale score in the observation group was (2.5 ± 0.8) points and (1.1 ± 0.5) points, respectively, which were significantly lower than (3.4 ± 0.9) points and (1.9 ± 0.8) points in the control group ( t = 5.01, -5.68, both P < 0.05). Conclusion:Oxycodone multimodal analgesia for laparoscopic gynaecological surgery can improve the quality of awakening, decrease systolic blood pressure, reduce the degree of postoperative pain, and does not increase the incidences of postoperative nausea and vomiting.