Objective: To analyze the chemical constituents of Polygonum multiflorum extract which may cause human liver cell damage and to explore the mechanism. Methods: Raw and processed Polygonum multiflorum were extracted by 70% ethanol, then raw and processed Polygonum multiflorum water-eluted material (RW and PW), 50% ethanol-eluted material (R50 and P50) and 95% ethanol-eluted material (R95 and P95) were obtained by absorbing through AB-8 macroporous resin, followed by water, 50% ethanol and 95% ethanol elution in order. The water extracts of raw and processed Polygonum multiflorum (RWE or PWE) were obtained by boiling them in water as usual. Normal human liver L02 cells were treated by different concentrations of eluted Polygonum multiflorum materials for different time, and the cell growth inhibition of each group was determined by methylthiazolyldiphenyl-tetrazolium bromide method. The chemical constituents which had a significant cytotoxicity to L02 cells were analyzed by high-performance liquid chromatography (HPLC). Morphological changes of L02 cells were observed by Giemsa staining and cell cycle distribution was observed by flow cytometry. Results: It was found that 95% ethanol-eluted extracts of raw and processed Polygonum multiflorum showed significant growth inhibition on normal human liver L02 cells, while the other components showed no significant inhibition on cell growth. HPLC analysis showed that the main component in 95% ethanol-eluted extract of raw and processed Polygonum multiflorum was emodin at content of (18.53+/-2.96)% and (10.28+/-1.34)% respectively. Cell cycle analysis showed that 95% ethanol-eluted material of Polygonum multiflorum and emodin had a similar significant effect of S phase arrest and all could induce L02 cell apoptosis. Conclusion: The main part of Polygonum multiflorum causing liver cell damage is the 95% ethanol-eluted extract, and emodin is one of the important chemical constituents leading to liver cell damage.

Objective　To establish a dual droplet digital PCR (ddPCR) assay for herpes simplex virus type I (HSV-1) and varicella-zoster virus (VZV). Methods The specific primers and probes were derived based on the conserved regions of HSV-1 and VZV genome. The primer-probe combinations were screened, and the annealing temperatures and primer-probe concentration ratios of the dual-droplet digital PCR reaction were optimized to establish a dual-droplet digital PCR reaction system for HSV-1 and VZV, which was tested for other viruses and validated for clinical samples. The sensitivity, specificity, and reproducibility of the established dual microtiter digital PCR method were analyzed. Results The optimal concentrations of primers and probes for the dual ddPCR detection method of HSV-I and VZV were determined to be 800 nmol/L and 250 nmol/L, respectively, with an optimal annealing temperature of 56 ℃. The correlation coefficient (R2) of the standard curve of the dual ddPCR assay was 0.99, showing a clear linear relationship. The method showed high sensitivity, with the lowest detection limit of herpes simplex virus type I being 2.97 copies/μL, and for VZV being 2.73 copies/μL. The repeatability was high with a small coefficient of variation and stable detection results; the specificity was excellent, and no cross-reaction was found with herpes simplex virus type Ⅱ, Epstein-Barr virus, Adenovirus, Coxsackievirus (CA6/CA10/CA16), Cytomegalovirus, Human Cytomegalovirus, Human enterovirus 71, Japanese Encephalitis virus, West Nile virus, Measles virus, Mumps virus, and human nucleic acids. Conclusions The dual droplet digital PCR assay for herpes simplex virus type I and varicella-zoster virus established in this experiment has strong sensitivity, specificity, and high repeatability, and can provide a solution for rapid quantitative detection of the two viruses in different scenarios.

ObjectiveTo study the relationship of the parental rearing behaviors between the psychological status and post-traumatic stress disorder in the injured deliberately.Methods 161 injured people were treated in five hospitals of the Urumqi and evaluated the self-reporting questionnaire-20 (SRQ20),7-items screening scale for PTSD( PTSD7 ),impact of event scale (IES) and symptom checklist 90 (SCL-90) in the third and the fourth week injured,including 113 injured people aged at 14 ～50 years old completed the Egna Minnen av Barndoms Uppfostran (EMBU).Two months after the injury,113 injured people were interviewed,including 106 injured people diagnosed by Structured Clinical Interview for DSM-Ⅳ-TR Axis Ⅰ Disorders/Patients (SCID-I/P) and 7 injured people were lost.ResultsFather's preferences correlated with the total score of IES ( r =- 0.234,P ＜ 0.05 ) and avoid factor( r=-0.309,P＜ 0.01 ) positively,Mother's emotional warmth and understand correlated with the flashback factor of IES ( r =0.194,P ＜ 0.05 ) negatively.The injured people were divided into three groups by diagnosis:the illness-free group,the PTSD group,and other group.Parental preferences correlated with Mental symptoms negatively.Father's punish severely,interference too much,refuse or deny and Mother's interference too much,over protection and punish severely correlated mental symptoms positively.The six subscales of father's rearing behaviors and the five subscales of mother's rearing behaviors had no significant difference.ConclusionsParental preferences in childhood can internalizes inside support to protect the psychological trauma in future.Father's punish severely,interference too much,refuse or deny and mother's interference too much,over protection and punish severely can damage mental health and aggravate symptoms after traumatic event.But the influence of parental rearing behaviors is limited to decide whether the injured people suffering from PTSD or other mental disorders.

Abstract： Objective　To elucidate the antigenic antibody reaction of recombinant expression of non-structural protein 1 (NS1) of Japanese encephalitis (JE) virus with various mosquito-borne flaviviruses, including JE virus, and the antigenic antibody reaction of serum samples of patients infected with JE virus in acute stage. Methods　In this study, Escherichia coli prokaryotic expression vector (pET) system was used to recombinant express Japanese encephalitis virus NS1 gene. Western Blot assay was performed to detect the antibody responses of the recombinantly expressed protein against a variety of mosquito-transmitted flaviviruses, including JE virus, as well as antigen-antibody reactions of serum from patients with acute JE virus infection. Results　The NS1 gene expression product of JE virus (P3 strain) was in the form of an inclusion body, and the denatured and renatured expression product was displayed as a single band in the denatured gel (polyacrylamide gel electrophoresis, PAGE), with a molecular weight of about 45 000. The results of further antigen-antibody analysis showed that the antigen/antibody hybridization reaction of the expression product with polyclonal or monoclonal antibody of JE virus (mosquito isolates, encephalitis isolates) and serum samples of patients with acute JE virus infection could be completely consistent. The recombinant product showed negative antigen/antibody hybridization reactions with mosquito-transmitted flaviviruses, such as dengue virus and yellow fever virus polyclonal antibodies, but positive reactions with polyclonal antibodies to West Nile virus and Murray Valley encephalitis virus. Conclusions　In this study, the recombinant expression of the NS1 protein of JE virus was successfully obtained, and the antigen/antibody reaction between the recombinant protein and samples of patients infected with mosquito-borne flavivirus and JE virus was analyzed. The study results provide important basic data for elucidating the antigen-antibody reaction between the NS1 protein of JE virus and mosquito-borne flavivirus. The recombinant expression protein obtained in this study provides an important material basis for further research on the function of JE virus NS1 protein.

Objective To explore the influencing factors of type 2 diabetes mellitus(T2DM)complicated with metabolic syndrome(MS)and investigate the correlation between the level of 25(OH)D3 and T2DM combined with MS.Methods 592 in patients with T2DM admitted to the Department of Endocrinology of Shenzhen Second People's Hospital were selected from January to December 2020.They were divided into simple T2DM group(T2DM,n=194)and T2DM combined with MS group(MS,n=398).Results Compared with the T2DM group,the MS group had lower 25(OH)D3 levels[26.30(20.50,32.39)vs 24.20(20.16,29.47),P<0.05].Spearman correlation analysis showed that 25(OH)D3 was negatively correlated with MS,FPG,TG,LDL-C,and 24 hUAlb(P<0.05 or P<0.01).Logistic regression analysis showed that 25(OH)D3 was a protective factor for T2DM combined with MS.Conclusions Patients with T2DM and MS have lower 25(OH)D3 levels.25(OH)D3 is a protective factor for T2DM combined with MS.

Objective:To understand the residents' cognition of commercial long-term insurance in Tianjin so as to provide a reference for improving service effect and quality of long-term insurance.Methods:A total of 31 residents from six districts of Tianjin were selected for semi-structured interviews from December 2022 to February 2023 by the convenient sampling method, and the data were analyzed by Colaizzi 7-step analysis method.Results:Three themes were extracted, namely, inadequate interpretation and promotion of commercial long-term insurance, immature development of commercial long-term insurance, and the ability of commercial long-term insurance to help caregivers reduce their family burden.Conclusions:The residents of Tianjin have a low awareness of commercial long-term insurance, which cannot meet the needs of most residents. It is urgent to strengthen their publicity further and improve their system.

Based on WeChat platform,blended teaching is designed and implemented in oral surgery practice.The undergraduate intern can get in touch with the internship early,become familiar with it early and make an internship program early.In the process of internship,they can achieve the purpose of getting more questions,more summary and more test through teaching,discussing and contesting.After the internship,they can consolidate,review,exchange and expand learning.The preliminary practice shows that the teaching stimulates the students' interest and motivation in the clinical study of oral surgery,enhances the interaction between teachers and students and the supervision of the teachers to the students,and improves the effect of the practice teaching.

Objective To investigate the relationship remifentanil-induced hyperalgesia and func-tion of nitrated glutamate transportor-1 ( GLT-1) and glutamine synthetase ( GS) in the spinal cord of rats with incisional pain. Methods Thirty-two male Sprague-Dawley rats, weighing 260-280 g, aged 2-3 months, in which caudal catheters were successfully implanted, were divided into 4 groups (n=8 each) u-sing a random number table method: control group ( group C) , incisional pain group ( group I) , remifen-tanil group ( group R) and remifentanil plus incisional pain group ( group RI) . Normal saline was intrave-nously infused for 60 min at 0. 1 ml · kg-1 · min-1 in group C. The model of incisional pain was estab-lished, and normal saline was simultaneously infused for 60 min via the tail vein at 0. 1 ml·kg-1 ·min-1 in group I. Remifentanil was infused for 60 min via the tail vein at 1. 0 μg· kg-1 ·min-1 in group R. The model of incisional pain was established, and remifentanil was infused for 60 min via the tail vein at 1. 0μg· kg-1 ·min-1 in group RI. The mechanical paw withdrawal threshold ( MWT) and thermal paw with-drawal latency ( TWL) were measured at 24 h before infusion of remifentanil or normal saline ( T0 ) and at 2, 6, 24 and 48 h after infusion ( T1-4 ) . The rats were sacrificed after the last measuremnet of pain thresh-old, and the L4-6 segment of the spinal cord was removed for determination of the expression of GLT-1 and GS (by Western blot) and expression of nitrated GLT-1 (nGLT-1) and nitrated GS (nGS) (by Western blot) . Ratios of nGLT-1∕GLT-1 and nGS∕GS were calculated. Results Compared with group C, the MWT was significantly decreased and TWL was shortened at T1-4 , the expression of GLT-1 and GS was down-regu-lated, the expression of nGLT-1 and nGS was up-regulated, and ratios of nGLT-1∕GLT-1 and nGS∕GS were increased in I, R and RI groups ( P<0. 05) . Compared with group I and group R, the MWT was signifi-cantly decreased and TWL was shortened at T1-4 , the expression of GLT-1 and GS was down-regulated, the expression of nGLT-1 and nGS was up-regulated, and ratios of nGLT-1∕GLT-1 and nGS∕GS were increased in group RI ( P<0. 05) . Conclusion The mechanism of remifentanil-induced hyperalgesia may be related to impaired function of GLT-1 and GS in the spinal cord of rats with incisional pain.

Objective:This study contrasts the immune efficacy of the varicella zoster virus glycoprotein E (VZV gE)using Al/CpG combined adjuvants and AS01 adjuvant in BALB/c mice.Methods:BALB/c mice were immunized at 0 and 21 days respectively, and serum antibodies were detected using enzyme-linked immunosorbent assay. Detection of neutralizing antibodies in mouse serum using varicella zoster virus; enzyme-linked immunosorbent spot assay was used to detect cellular immune response.Results:Following two intramuscular immunizations, mice in the experimental groups (Shingrix, gE+ Al/CpG, and gE+ AS01) demonstrated elevated neutralizing antibody titers and an augmented count of lymphocytes releasing IFN-γ and IL-4. The gE+ Al/CpG group displayed the highest neutralizing antibody titer (1943), yet the AS01-adjuvanted groups (Shingrix and gE+ AS01) showed increased lymphocyte counts secreting IFN-γ and IL-4 compared to the Al/CpG group (gE+ Al/CpG). In comparison to the AS01 adjuvant, Al/CpG adjuvants triggered a humoral immune response favoring Th2 in mice. The proportions of CD4 + T and CD8 + T cells were not significantly different among the experimental groups. Conclusions:Al/CpG adjuvant combined with gE protein resulted in high neutralizing antibody titers, while the intensity of the induced cellular immune response was inferior to that of AS01 adjuvant.

Objective To investigate the influencing factors for white matter hyperintensity （WMH） in middle-aged and elderly patients with hypertension， and to establish and verity a nomogram prediction model. Methods A total of 198 middle-aged and elderly patients with hypertension and WMH who were hospitalized in our hospital from January 2022 to April 2023 were enrolled. Related clinical data were analyzed， and related data were recorded. A binary logistic regression analysis was used to investigate the independent risk factors for WMH and establish a nomogram， and the receiver operating characteristic （ROC） curve and the calibration curve were used to evaluate the diagnostic efficacy of the nomogram. Results Age， course of hypertension， cystatin C， homocysteine，red blood cell distribution width， and cognitive impairment were the independent influencing factors for WMH in the middle-aged and elderly patients with hypertension. The nomogram established showed good diagnostic efficacy （AUC=0.815， 95% CI 0.756~0.874，P<0.001） and calibration ability （C index=0.794）. Conclusion The nomogram established in this study has a good predictive ability for moderate-to-severe WMH in middle-aged and elderly patients with hypertension and can provide certain help for clinical workers.
Nomograms