Objective To explore the role of extracellular signal-regulated kinase (ERK)in central nucleus of amygdala (CeA)in the mechanism of fentanyl-induced hyperalgesia (OIH)in rats. Methods Step 1:12 healthy male Sprague-Dawley rats,weighing 60-100 g,were randomly divided into OIH and Control group.The mechanical paw withdrawal threshold (PWT)and the thermal paw withdrawal latency (PWL)were tested at pre-and post-OIH induction.Then the level of p-ERK in the CeA was analyzed by Western blotting.Step 2:After successful induction of OIH and catheterization in CeA,another 30 SD male rats were randomly divided into 5 groups (n = 6 each):Group OIH;Group OIH+U0124;Group OIH+U0126(0.1 5 nmol);Group OIH+U0126(0.45 nmol)and Group OIH+U0126 (1.5 nmol),then 0.3 μl of DMSO,U0124 (1.5 nmol),U0126 (0.1 5 nmol,0.45 nmol,1.5 nmol)was given through the catheter separately.PWT and PWL were tested before cathe-terization,at pre-OIH induction,post-OIH induction and 0.5 h after CeA drug administration.After the last test of pain threshold,the rats were sacrificed and CeA tissues were sampled for analyzing the expression of p-ERK by western blot.Results In step 1 compared with control group,PWT and PWL of OIH group were sharply decreased post-OIH induction (P <0.05),concomitant increase of the expression of p-ERK in CeA in OIH group was also observed.In step 2,both PWT and PWL were sharply decreased post-OIH induction (P <0.05).Intra-CeA U0126 injection,but not U0124, reversed both behavioral hyperalgesia and molecular activation of ERK in CeA in a dose-dependent manner (P <0.05).Conclusion ERK plays a pivotal role in the maintenance of fentanyl-induced hy-peralgesia.Targeting inhibition of ERK activation in CeA can alleviate fentanyl-induced hyperalgesia.

Objective To investigate the role of extracellular signal-regulated kinase1/2 (ERK1/2) in the central amygdala on fentanyl-induced hyperalgesia in rats.Methods Thirty-two male SpragueDawley rats, weighing 60-100 g, were randomly divided into 4 groups (n =8 each) using a random number table: control group (group C), fentanyl-induced hyperalgesia group (group H), U0124 group (group U1) , and U0126 group (group U2).A catheter was implanted in the central amygdale.In group C, normal saline was injected subcutaneously, and 6.5 h later dimethyl sulfoxide (DMSO) was injected via the catheter.In group H, fentanyl was injected subcutaneously to induce hyperalgesia, and 6.5 h later DMSO was injected via the catheter.In group U1, hyperalgesia was induced, and 6.5 h later ERK1 inhibitor U0124 1.5 nmol was injected via the catheter.In group U2, hyperalgesia was induced, and 6.5 h later ERK1/2 inhibitor U0126 1.5 nmol was injected via the catheter.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal threshold (TWT) were measured before fentanyl injection, at 6.5 h after injection, and at 30 min after DMSO or U0124/U0126 administration via the catheter (T0-2).After the last measurement of pain threshold, the rats were sacrificed, and the amygdala tissues were sampled for detection of the expression of phosphorylated ERK1/2 (p-ERK1/2) by Western blot in groups C and H.Results Compared with group C, the MWT and TWT were significantly decreased at T1,2in H and U1 groups, and at T1in group U2 (P＜0.05) , the expression of p-ERK2 was up-regulated (P＜0.05) , and no significant change was found in the expression of p-ERK1 in group H (P＞0.05).Compared with group H,the MWT and TWT were significantly increased at T2 in group U2 (P＜0.05) , and no significant change was found in MWT, TWT in group U1 (P＞0.05).Conclusion ERK2 activation in the central amygdala is involved in the development of fentanyl-induced hyperalgesia in rats.

Objective:To study the effect of overdose fluoride on Smad3 expression during development course of Wistar rat's incisor,and then to explore pathogenesis of dental fluorosis.Methods:20 Wistar rats(10 male rats and 10 female rats)were randomly divided into two groups.Experiment group was feed with water contained 0.1 g/L fluorin and normal group with distilled water was served as control.After 8 weeks all rats were sacrificed.Immunohistochemistry was used to observe the effect of overdose fluorin on Smad3 expression in rat's ameloblast and odontoblast.Results:The immunohistochemistry results demonstrated that in experiment group,Smad3 expression in rat's ameloblast and odontoblast was lower than those in control group,and statistic analysis showed there were significant differences between the two groups(P

Objective : To identify the factors affecting the adherence to administration of nucleoside/nucleotide analogues among patients with chronic hepatitis B (CHB), so as to provide insights into improving medication adherence among CHB patients. Methods: CHB patients at ages of 18 to 44 years that underwent treatment in Hangzhou First People's Hospital Affiliated to Zhejiang University School of Medicine from June 2021 to December 2022 were enrolled, and subjects' demographic and clinical characteristics were collected. The medication adherence was evaluated with the Morisky Medication Adherence Scale, and the negative illness perception was evaluated using the simplified Illness Perception Questionnaire, while the social support level was evaluated with the Social Support Scale. In addition, factors affecting the medical adherence were identified using a multivariable logistic regression model among CHB patients. Results: A total of 225 questionnaires were allocated and 210 valid questionnaires were recovered, with an effective recovery rate of 93.33%. The respondents had a mean age of (35.94±8.72) years, and included 119 men (56.67%) and 91 women (43.33%), with a mean total illness perception score of (44.09±12.06) points. Morisky Medication Adherence Scale classified good in 149 cases (70.95%) and poor in 61 cases (29.05%). Multivariable logistic regression analysis identified an educational level of college degree and above (OR=1.676, 95%CI: 1.105-2.910), family monthly per capita income of >5 000 Yuan (OR=2.518, 95%CI: 1.360-5.033), receiving health education pertaining to CHB (OR=3.590, 95%CI: 1.611- 7.932), high or medium level of social support (high: OR=3.202, 95%CI: 1.613-5.870; medium: OR=1.267, 95%CI: 1.085-2.409) as factors improving the medication adherence, and administration of more than 3 types of drugs (OR=0.388, 95%CI: 0.204-0.526) and high illness perception score (OR=0.601, 95%CI: 0.395-0.808) as factors constraining medication adherence among CHB patients. Conclusions The medication adherence are affected by educational level, family monthly per capita income, types of drugs, illness perception score, health education pertaining to CHB and social support level among CHB patients. Individualized intervention strategy is required targeting these influencing factors to improve the medication adherence among CHB patients.

Objective To observe the intervention of sub-health population with modified Xiaoshengliu Decoction, a Chinese herbal recipe with the actions of strengthening Qi and nourishing yin, soothing liver and relieving stagnation, tonifying liver and kidney. Methods The clients of professor LUO Ren from Dept. of Traditional Chinese Medicine, Nanfang Hospital Affiliated to Southern Medical University, were chosen as the studying objects. Sub-health outpatient cases were defined according to the data of four diagnostic methods together with Sub-health Questionnaire developed by the sub-health program of Southern Medical University, and then were given modified Xiaoshengliu Decoction (mainly composed of Radix Bupleuri, Radix Scutellariae, Rhizoma Pinelliae Preparata, Radix Codonopsis, Radix Rehmanniae Preparata, Rhizoma Dioscoreae, Fructus Corni, Cortex Moutan, Radix Ophiopogonis, Fructus Schisandrae Chinensis, Radix Glycyrrhizae Preparata, etc.) for intervention. The symptoms of the subjects were followed up before and after treatment for the evaluation of therapeutic effect with Sub-health State Symptom Checklist. Results (1) The results showed that the total effective rate was 92.03%at first follow -up, and was 100% at the last follow-up, the difference being significant between the two periods of follow-up (P<0.01) . (2) The results of single-factor variance analysis showed that the differences of symptoms scores before treatment, at the first and the last follow-up were statistically significant (P<0.01) . The symptoms scores before treatment, at the first follow-up and at the last follow-up were in decreasing sequence (P<0.01) . Conclusion Modified Xiaoshengliu Decoction shows certain effect on the intervention of sub-health population with the syndrome of Qi-yin deficiency .

Objective To investigate the low hemoglobin density (low haemoglobin density,LHD) and red blood cell volume distribution width (red blood cell volume distribution width,RDW) in iron deficiency anemia (iron-deficiency anemia,IDA)in children with the value of diagnosis and treatment.Methods From February 2016 to May 2017,in the Second People's Hospital of Longgang District in Shenzhen City,86 cases of children with iron deficiency anemia for IDA group,and 120 cases of healthy children (as the control group) at the same time were confirmed.Blood routine of children with IDA were detected before and after treatment hemoglobin concentration and the results were analyzed statistically.Results 120 cases of healthy children in the peripheral blood LHD value was 2.74 ％ ± 0.90 ％ and the boy was 3.07 ％ ± 0.81％,higher than that of 2.26 ％ ± 0.69 ％ of the girls.Between them,there was statistically significant difference (t=3.815,P＜0.05).The value of RDW was 12.37 ％ ± 2.84 ％,12.09 ％ ± 2.80 ％ for the boys and 12.56 ％ ± 2.87 ％ for the girls,there was no statistically significant difference between the them (t=0.517,P＞0.517).Before treatment,86 cases of children with IDA LHD and RDW values in peripheral blood were 30.67 ％ ± 20.12 ％ and 16.62 ％ ± 3.27 ％,significantly higher than the control group,the difference was statistically significant between (t=4.025 ～ 4.025,P＜0.05 ～ 0.01),and there was no statistically significant difference between male and female children (t =0.761 ～ 0.917,P＞ 0.05).After treatment L HD and RDW values were 10.65 ％ ± 8.92 ％ and 14.02 ％ ± 2.93 ％,significantlylower than before the treatment,between them was statistically significant difference (t=2.912～9.675,P＜0.05).Conclusion Children with iron deficiency anemia treatment before the LHD and RDW values significantly elevated in the blood,significantly decreased after treatment,the LHD and RDW values for diagnosis and treatment of children with iron deficiency anemia dynamic monitoring has certain application value,worthy of popularization and application.

OBJECTIVETo determine the effect of type I transmembrane protein (IRE1alpha) deletions on the cell cycle of human periodontal ligament fibroblasts (hPDLFs) cells.

METHODSBased on the IRE1alpha deletions, a full-length model was successfully constructed. Moreover, overlapping polymerase chain reaction mutagenesis facilitated the establishment of two deletion mutants of IREla (pD-Kinase, pD-Rnase). The full-length model and two mutant eukaryotic expression vectors were transfected into hPDLFs cells. Western blot analysis was performed to identify the expression in the cells. The changes in the cell cycle of hPDLFS cells were detected by flow cytometry (FCM).

RESULTSThe two deletion mutants of IRE1alpha with eukaryotic expression vectors were successfully constructed and correctly expressed in hPDLFs cells based on Western blot analysis. Under stress conditions, the FCM assay showed that cell percentage of S phases increased, whereas that of G1 phases decreased in the IRE1alpha group (P < 0.05) compared with the control group of tunicamycin (TM) treatment. Moreover, the cell percentage of the S phases decreased, whereas that of the G1 phases increased in the D-Rnase group (P < 0.05) compared with the control. The deletion mutant D-Kinase had no influence on hPDLFS cell proliferation and cycle (P>0.05).

CONCLUSIONUnder stress conditions, IRE1alpha can improve the cell cycle of hPDLFs cells from the G1 to the S phase. The deletion mutant D-Rnase cause hPDLFs cell growth arrest at the G1 phase, whereas deletion mutant D-Kinase has no significant effect.

Objective To investigate the effect of gene silencing of Y-box binding protein-1(YB-1) by RNA interference on the proliferation and migration in prostate cancer cells lines PC-3 cells .Methods YB-1 siRNAs(pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2) were synthesized and transfected into cloned into the the PC-3 cells by liposome .The expressions of YB-1 were measure by RT-PCR and Western blotting .The proliferation and migration were respectively detected by MTT and Transwell method . Results ThemRNA and protein expressions of YB-1 were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2 (P<0 .05) ,compared with the control group ,the inhibition ratio of mRNA expression was 36 .23% and 39 .42% respectively and the inhibition ratio of protein expression was 41 .56% and 55 .33% respectively .The proliferation and migration were significantly decreased by pGenesil-1-YB-1-1 and pGenesil-1-YB-1-2(P<0 .05) .Conclusion YB-1 gene silencing by RNA interference inhibits the proliferation and migration in prostate cancer cells lines PC-3 cells .

Objective: To investigate the perception about illness and identify its influencing factors among patients with non-alcoholic fatty liver disease ( NAFLD ) , so as to provide insights into the management of NAFLD patients. Methods: NAFLD patients admitted to Hangzhou First People's Hospital Affiliated to Medical School of Zhejiang University from January to June, 2020, were selected as the study subjects, and subjects' demographic features were collected using questionnaires, including gender, age and education level. The perception about illness, coping models and social support were assessed using the Brief Illness Perception Questionnaire (BIPQ), Medical Coping Modes Questionnaire ( MCMQ ) and Social Support Rating Scale ( SSRS ), respectively, and factors affecting the perception about illness were identified using multivariable linear regression analysis among NAFLD patients. Results: The 286 respondents included 151 males ( 52.80% ) and 135 females ( 47.20% ), and had a mean age of ( 55.27±10.39 ) years. The mean illness perception score was 38.55±9.21 among the respondents. The mean SSRS score was 42.90±8.64. The mean coping mode scores of confronce, avoidance and resignation were 23.51±4.30, 17.49±2.82, and 7.12±2.05, respectively. Multivariable linear regression analysis identified education level ( high school, β'=-0.216; diploma and above, β'=-0.355 ), household monthly income per capita ( β'=-0.372 ), regular exercise ( β'=-0.310 ), coping modes ( confronce, β'=-0.326; avoidance, β'=-0.191 ) and social support level ( β'=-0.259 ) as factors affecting the perception about illness among NAFLD patients. Conclusion Negative perceptions about illness are found among NAFLD patients, and household income, education level, regular exercise and coping modes are factors affecting the illness perception among NAFLD patients.

Objective: To investigate immune responses to influenza virus infections and the immunogenicity of trivalent split-virus influenza vaccine among elderly populations in Jiangbei District, Ningbo City, so as to provide the support for promoting influenza vaccination among elderly populations. Methods: The elderly populations at ages of 60 years and older were recruited in Jiangbei District of Ningbo City from September to November, 2020, and the participants were assigned to the vaccination group and the control group according to vaccination intention. The titers of haemagglutination inhibition ( HI ) antibodies against influenza viruses A ( H1N1 and H3N2 ) and BV were measured using the micro HI test prior to vaccination and 30 days post-vaccination, and the protective rate, geometric mean titer ( GMT ) and seroconversion rate of antibodies were analyzed before and after vaccination. Results: There were 290 participants in the vaccination group, including 132 men (45.52% ), and 290 controls, including 132 men ( 45.52% ). There were no significant differences between the vaccination group and the control group in terms of the protective rate or GMT of antibodies against influenza viruses A ( H1N1 and H3N2 ) and BV prior to vaccination ( P>0.05 ). Following vaccination, the protective rates of antibodies against influenza viruses A ( H1N1 and H3N2 ) and BV were 98.62%, 94.14% and 88.28%, and the GMT of antibodies against influenza viruses A ( H1N1 and H3N2 ) and BV increased by 9.26, 6.19 and 10.09 folds, while the seroconversion rates of antibodies against influenza viruses A ( H1N1 and H3N2 ) and BV were 78.62%, 68.28% and 71.38%, respectively. The protective rates, GMT and seroconversion rates of antibodies against influenza viruses A ( H1N1 and H3N2 ) and BV were all significantly greater in the vaccination group than in the control group post-vaccination ( P<0.05 ). A lower increase was seen in the GMT of antibodies against the influenza virus BV among residents at ages of 80 years and older (increase by 7.91 folds) than among residents at ages of 70 to 79 years ( increase by 12.53 folds ) and 60 to 69 years (increase by 13.32 folds) in the vaccination group post-vaccination ( P<0.05 ), and the seroconversion rate of antibodies against the influenza virus BV was significantly lower in residents at ages of 80 years and older ( 62.57% ) than in those at ages of 70 to 79 years ( 83.33% ) ( P<0.05 ), while the positive conversion rate of antibodies against the influenza virus A ( H3N2 ) was significantly lower in residents at ages of 80 years and older ( 62.57% ) than in those at ages of 60 to 69 years ( 91.30% ) ( P<0.05 ). Conclusions Low-level immune responses are detected to antibodies against influenza virus A ( H3N2 ) and BV among elderly populations in Jiangbei District of Ningbo City, and trivalent split-virus influenza vaccine shows a high immunogenicity among elder populations. An emphases on improvements in coverage of influenza vaccination among elderly populations at ages of 60 to 69 years, and development of influenza vaccines with a higher protective efficacy for residents at ages of 80 years and older are recommended.