African swine fever virus(ASFV)I177L gene deletion vaccine is one of the key directions of African swine fever(ASF)live attenuated vaccine research and development.In order to effec-tively distinguish between the wild-type ASFV strain and the I177L gene-deleted strain,specific primers and probes were designed based on ASFV B646L and I177L genes,respectively.After screening and optimization,a duplex real-time PCR method was developed that can simultaneously detect these two genes.The results showed that ASFV B646L and I177L genes were detected spe-cifically and simultaneously by the method developed without cross-reactions with porcine circovir-us type 2,Seneca virus A,classical swine fever virus,foot-and-mouth disease virus,porcine respira-tory and reproductive syndrome virus.The detection limits of the duplex real-time PCR for recom-binant plasmids pUC57-B646L and pUC57-I177L were 1×103 copies/mL.The intra-and inter-as-say coefficients of variation were less than 4％,respectively.Detection of 122 pork and pork prod-ucts using the duplex real-time PCR developed and the real-time PCR recommended by WOAH showed that the coincidence rates of the two methods for B646L gene detection was 100％with two amplification curves appeared in the positive results of the established methods.The method established in this study can be used for the detection of ASFV I177L gene deletion strains,which provides technical support for ASF surveillance and epidemiological investigation.

Objective:To summarize the clinical features and genetic characteristics of Congenital bile acid synthetic disorder type 3 (BASD3) disorder caused by CYP7B1 gene variation.Methods:This was a case series study. Clinical data and genetic results of 2 cases of congenital bile acid synthetic disorder type 3 caused by CYP7B1 gene variations in the Department of Infectious Diseases, Children′s Hospital of Fudan University at Xiamen and Department of Pediatrics, Women and Children′s Hospital, School of Medicine, Xiamen University from January 2021 to December 2023 were retrospectively collected and analyzed. Literature up to December 2023 was searched from electronic databases of China National Knowledge Infrastructure (CNKI), Wanfang Data and PubMed with the combined keywords of " Congenital bile acid synthetic disorder type 3""Oxysterol 7-alpha-hydroxylase""Oxysterol 7α-Hydroxylase Deficiency""BASD3" and "CYP7B1 liver" both in Chinese and English. The main clinical features and genetic characteristics of BASD3 disorder caused by CYP7B1 gene variations were summarized.Results:Two BASD3 patients, 1 male and 1 female, were admitted at the ages of 3 months and 18 days, and 2 months and 7 days, respectively. Both patients presented with neonatal cholestasis and hepatomegaly. Biochemical evidence indicated direct hyper-bilirubinemia with elevated aminotransferase levels, while gamma-glutamyltransferase (GGT) and total bile acid levels were normal or nearly normal. Patient 1 was a compound heterozygotes of the CYP7B1 gene variants c.525-526insCAAGTTGG(p.Asp176GInfs*15) and c.334C>T(p.Arg112Ter). Patient 1 jaundice resolved and liver function tests normalized after oral administration of chenodeoxycholic acid (CDCA). Patient 2 was homozygous for variant c.334C>T(p.Arg112Ter) in CYP7B1 gene. Patient 2 was in liver failure status already and not reactive to oral CDCA administration. Patient 2 received living-related liver transplantation for enhanced abdominal CT revealed a liver tumor likely vascular origin. Literature review revealed no cases of BASD3 reported in Chinese literature, including 2 patients in this study, while 12 patients (9 males and 3 females) were reported in 9 English literatures. All of the 12 manifested jaundice and hepatosplenomegaly in infancy, with cirrhosis, liver failure, kidney enlargement, hypoglycemia, and spontaneous bleeding in some cases, polycystic kidney disease was demonstrated in 5 cases of them. The c.334C>T (p.Arg112Ter) of the CYP7B1 gene was homozygous in 4 cases and compound heterozygous in 2 cases. Among the 12 children, 6 cases received CDCA treatment, while 6 cases not. Four survived with their native liver in the 6 cases who received CDCA therapy, while none in the 6 cases not received CDCA therapy.Conclusions:BASD3 is a rare hereditary cholestatic disorder. Markedly elevated levels of conjugated bilirubin and aminotransferases, with normal or nearly normal GGT and total bile acid levels can serve as diagnostic clue. c.334C>T is the most common pathogenic variant of the CYP7B1 gene. Timely administration of CDCA may save the liver.

Objective:To explore the efficacy of online problem management plus (PM+) intervention on the mental health among adults with anxiety.Methods:Ninety subjects with anxiety (Generalized Anxiety Disorder-7 (GAD-7) total score≥5) were enrolled and randomly allocated into either waiting group or online PM+group. Participants in the online PM+intervention group received online PM+intervention twice a week for 3 weeks, while participants in the waiting group received general psychological supports. Psychological evaluation was performed at the end of the 3-week treatment and at 6 months after treatment. Outcome measures included GAD-7, Patient Health Questionnaire -(PHQ-9), Patient Health Questionnaire-15 (PHQ-15), Perceived Stress Scale-14 (PSS-14), and Insomnia Severity Index (ISI). Two-factor repeated measure analysis of variance (ANOVA) was used to compare the scores of the two groups at baseline and after intervention. Single-factor repeated measure analysis of variance was used to compare the differences of scores at baseline,3-week post-intervention, and 6-month follow-up in the online PM+ group.Results:A total of 37 (37/45) pations in the online PM+intervention group and 30 (30/45) patients in the waiting group completed the psychological evaluation after intervention. After 3-week intervention, compared with baseline, no significant change was found in the scores of GAD-7 ( F=0.08, P=0.782), PHQ-9 ( F=0.33, P=0.570), PHQ-15 ( F=0.20, P=0.660), PSS-14 ( F=0.14, P=0.05) and ISI ( F=0.02, P=0.880) in the waiting group. The changes of GAD-7 ( F=22.61, P<0.001), PHQ-9 ( F=19.49, P<0.001), PHQ-15 ( F=12.67, P=0.001), PSS-14 ( F=16.69, P<0.001) and ISI ( F=5.59, P=0.022) scores in the online PM+group were statistically significant. There were significant differences in GAD-7 (9.7±5.2 vs. 5.0±3.5, F=17.11, P<0.001), PHQ-9 (11.4±5.9 vs. 6.9±4.7, F=11.65, P=0.002), PHQ-15 (10.4±5.4 vs. 6.3±4.1, F=12.24, P=0.002) and PSS-14 (26.0±7.5 vs.31.8±9.9, F=6.59, P=0.016) scale scores between the online PM+group and the waiting group after intervention. In addition, the scores of GAD-7 (95% CI=1.25-6.47, P=0.002) and PHQ-9 (95% CI=2.21-9.10, P=0.005) scales in the online PM+group still had statistically significant differences compared to the baseline at the 6-month follow-up. Conclusions:In this study, online PM+intervention significantly alleviated symptoms of anxiety, depression, somatization, stress, and insomnia in adults, and the therapeutic benefits of PM+persisted for at least 6 months.

Objective Observe the effect of Liuwei Anshen Capsule on sleep improvement of aged sleep deprivation zebrafish model.Methods Sixty 18-month-old female zebrafish were randomly divided into blank group,model group,Liuwei Anshen group,and melatonin group.The zebrafish in the blank group were raised under normal lighting conditions,and the other three groups were constructed with continuous lighting for 3 days.Zebrafish in Liuwei Anshen group were treated with Liuwei Anshen capsule water solution 0.000 50 mg·mL-1 for 3 days on the basis of model group,and zebrafish in melatonin group were treated with melatonin aqueous solution 0.2 mg·mL-1 on the basis of model group for 3 days.After 3 days,the zebrafish behavior system was used to detect the resting time of zebrafish in each group.qRT-PCR method was used to detect cyclin 1a(per1a),cyclin2(per2),circadian motor output cycle 1a(clock1a),cryptochrome 1b(cry1b),and 5-hydroxyindoleacetic acid(5-htiaa)in each group of zebrafish and follicle-stimulating hormone beta(fshβ)gene expression levels.Western blot was used to detect the expression levels of estrogen receptorα(Esrα),Fshβand luteinizing hormone(Lh)in zebrafish in each group.HE staining was used to observe the ovary of zebrafish in each group.Results Compared with the zebrafish in the blank group,the resting time of the zebrafish in the model group decreased significantly(P<0.01)during the 24 hours of observation.After the intervention of Liuwei Anshen Capsule and melatonin,the resting time of the zebrafish was significantly increased.(P<0.01).Compared with the zebrafish in the blank group,the mRNA expressions of zebrafish circadian clock genes per1a,per2,clock1a,cry1b,5-htiaa,and fshβ all showed a downward trend after sleep deprivation(P<0.05,P<0.01).After the intervention of Liuwei Anshen Capsules,the mRNA expressions of per1,clock1a,cry1b,5-htiaa and fshβ were all up-regulated(P<0.05,P<0.01).Compared with the zebrafish in the blank group,the protein expressions of Esrα and Lh in the zebrafish of the model group were up-regulated(P<0.05),the expression of Fshβ protein was down-regulated(P<0.05),after the intervention of Liuwei Anshen Capsules,the above proteins did not change significantly.The ovarian tissue cells of the zebrafish in the blank group had normal morphology and a large number of primary oocytes,while the ovarian tissue cells of the zebrafish in the model group were damaged in morphology and the number of primary oocytes decreased,after the intervention of Liuwei Anshen Capsule and melatonin,the cell morphology of zebrafish ovarian tissue was still damaged to varying degrees,but the whole was relatively intact,and the number of primary oocytes increased.Conclusion The insomnia of aged zebrafish may be caused by multiple factors.Liuwei Anshen capsule has significant effects on estrogen level of rhythm gene and ovarian function of sleep-deprived aged zebrafish.

Objective:To explore the efficacy of online problem management plus (PM+) intervention on the mental health among adults with anxiety.Methods:Ninety subjects with anxiety (Generalized Anxiety Disorder-7 (GAD-7) total score≥5) were enrolled and randomly allocated into either waiting group or online PM+group. Participants in the online PM+intervention group received online PM+intervention twice a week for 3 weeks, while participants in the waiting group received general psychological supports. Psychological evaluation was performed at the end of the 3-week treatment and at 6 months after treatment. Outcome measures included GAD-7, Patient Health Questionnaire -(PHQ-9), Patient Health Questionnaire-15 (PHQ-15), Perceived Stress Scale-14 (PSS-14), and Insomnia Severity Index (ISI). Two-factor repeated measure analysis of variance (ANOVA) was used to compare the scores of the two groups at baseline and after intervention. Single-factor repeated measure analysis of variance was used to compare the differences of scores at baseline,3-week post-intervention, and 6-month follow-up in the online PM+ group.Results:A total of 37 (37/45) pations in the online PM+intervention group and 30 (30/45) patients in the waiting group completed the psychological evaluation after intervention. After 3-week intervention, compared with baseline, no significant change was found in the scores of GAD-7 ( F=0.08, P=0.782), PHQ-9 ( F=0.33, P=0.570), PHQ-15 ( F=0.20, P=0.660), PSS-14 ( F=0.14, P=0.05) and ISI ( F=0.02, P=0.880) in the waiting group. The changes of GAD-7 ( F=22.61, P<0.001), PHQ-9 ( F=19.49, P<0.001), PHQ-15 ( F=12.67, P=0.001), PSS-14 ( F=16.69, P<0.001) and ISI ( F=5.59, P=0.022) scores in the online PM+group were statistically significant. There were significant differences in GAD-7 (9.7±5.2 vs. 5.0±3.5, F=17.11, P<0.001), PHQ-9 (11.4±5.9 vs. 6.9±4.7, F=11.65, P=0.002), PHQ-15 (10.4±5.4 vs. 6.3±4.1, F=12.24, P=0.002) and PSS-14 (26.0±7.5 vs.31.8±9.9, F=6.59, P=0.016) scale scores between the online PM+group and the waiting group after intervention. In addition, the scores of GAD-7 (95% CI=1.25-6.47, P=0.002) and PHQ-9 (95% CI=2.21-9.10, P=0.005) scales in the online PM+group still had statistically significant differences compared to the baseline at the 6-month follow-up. Conclusions:In this study, online PM+intervention significantly alleviated symptoms of anxiety, depression, somatization, stress, and insomnia in adults, and the therapeutic benefits of PM+persisted for at least 6 months.

With the development of detection methods, various biomarkers of liver cancer have been detected constantly, which is of great significance for the early diagnosis and real-time monitoring of liver cancer after treatment. Based on the differences in the sensitivity and specificity of different biomarkers, exploration of the value of diverse biomarkers in the diagnosis and prognosis assessment of liver cancer can provide an important reference for clinicians to scientific and rational application of distinct biomarkers.

The pregnancy registry for medicine is a common method for risk evaluation for drug safety evaluation during pregnancy. The authors introduced the exploration and practice of the pregnancy drug registry mode based on pharmacy service in Women′s Hospital School of Medicine Zhejiang University. The registry of pregnancy medication with the drug consultation clinic as the fulcrum was managed by the pharmacist team in a homogenization way, and implemented according to the information process of consultation, pharmacy guidance and regular follow-up. In the consultation and follow-up work, the pharmacists established the consultation drug history through three data sources: independent report of the consultant, inquiry of pharmaceutical personnel and case sampling. The pharmacists designed and constructed information screening system, classified the consultation medicine history according to the pregnancy exposure of specific drugs. The prospective research method was designed to meet the characteristics of pregnancy medication. The study was carried out on the relationship between pregnancy exposure and birth defects of offspring. Relying on the improvement of standardization, refinement and information management level of pharmaceutical services, multi center cooperation will be strengthened in the future to carry out continuous research on pregnancy drug exposure registry and follow-up system.

Objective:To retrieve, appraise and summarize the evidence for the airway humidification management of the inpatient with laryngectomy and tracheotomy but nonmechanical ventilation and to provide references for the clinical nursing.Methods:The evidence question was raised based on the study objective. The evidence was retrieved from some databases according to the evidence pyramid model during inception to December, 2018. The literature types included clinical practice guideline, evidence summary, best practice information sheet, recommended practice and systematic review. The quality of the literature were evaluated by the suitable evaluated tool based on their types. The level and recommedation grade of the evidence were appraised by the suitable tools of JBI.Results:Thirteen studies were recruited, including one clinical practice guideline, two evidence summaries, one best practice information sheet and nine systematic reviews. Totally thirteen items of best evidence were summarized and generalized to four categories including assessment, method, liquid and operation of airway humidification.Conclusion:The evidences of the studies are scientific and practical, but on the one hand, it is recommended that when applying the evidences in clinic, it is necessary to assess the clinical situation and chose the proper evidence. And on the other hand, there are lacking of some evidences of airway humidification and the level of some evidences is low. So it needs to create more high level evidences.

[Abstract] Objective: To investigate the effect of alantolactone (ALT) on proliferation, migration, invasion and apoptosis of human osteosarcoma 143B cells and the underlying mechanism. Methods: Osteosarcoma 143B cells were treated with different concentrations of ALT (0, 4, 6, 8, 10 µmol/L). Then, the cell proliferation ability was detected by crystal violet staining and MTT assay, cell migration was determined by Wound-healing test, cell invasion was analyzed by Transwell assay and cell apoptosis rate was detected by Hoechst33258 staining. The mRNA and protein expressions of E-cadherin, N-cadherin, caspase-3, cleaved caspase-3 (c-caspase-3), poly ADP-ribose polymerase (PARP) and cleaved PARP (c-PARP) in 143B cells were detected by qPCR and Western blotting (WB), respectively. TCF/LEF (T cell lymphocyte factor/lymphoid enhancer factor) transcriptional activity was examined with Luciferase reporter gene assay. The mRNA and protein expressions of β-catenin as well as MMP-7 and c-Myc were detected by qPCR and WB, respectively. Results: ALT inhibited proliferation, migration and invasion of osteosarcoma143B cells and promoted apoptosis(P<0.05or P<0.01). After the treatment with ALT at 8, 10 µmol/L, the mRNA and protein expressions of E-cadherin and PARP, as well as the protein expressions of c-caspase-3 and c-PARP were up-regulated, while the mRNA and protein expressions of N-cadherin were downregulated (P<0.05 or P<0.01);At the sametime, theTCF/LEF transcriptional activity and the mRNA and protein expressions of β-catenin, MMP-7 and c-Myc were significantly down-regulated (P<0.05 or P<0.01). Conclusion:ALT may inhibit the proliferation, migration and invasion and promote cell apoptosis possibly through suppressing Wnt/β-catenin signaling pathway in osteosarcoma 143B cells.

Jervine, a novel steroidal alkaloid from Veratrum nigrum L., exhibits both antitumor effect and potential toxicity. The aim of study was to characterize the pharmacokinetic behaviors and enterohepatic circu-lation of jervine in rats. A rapid and simple ultra-high performance liquid chromatography-tandem mass spectrometric method was developed and validated for quantification of jervine and alpinetin (internal standard) in rat plasma. After extraction from rat plasma by a simple protein-precipitation method, the analyte was separated on a C18 column (2.1 mm × 50 mm, 1.7μm) using water with 0.1％formic acid and acetonitrile as the mobile phase delivered at a flow rate of 0.4 mL/min. Jervine and alpinetin were determined in the positive mode with multiple reaction monitoring (MRM) of the ion transitions at m/z 426.3→108.8 and m/z 271.0→166.9, respectively. Molecular docking method was used to investigate the binding of jervine to p-glycoprotein and dehydroepiandrosterone sulfotransferase. The method was well validated within acceptance limits including specificity, matrix effect, recovery, precision, accuracy, and stability, and was successfully applied to the pharmacokinetic study of jervine after oral and intravenous administration to rats. Jervine presented a small volume of distribution, fast absorption, high oral bioavailability, and enterohepatic circulation. The enterohepatic circulation was first observed in veratrum alkaloids, and was further investigated by molecular docking studies, which was related to the binding of jervine to p-glycoprotein and dehydroepiandrosterone sulfotransferase. The pharmacokinetic properties and enterohepatic circulation of jervine in rats provided a significant basis for the drug-drug interaction and toxicity study in the future.