ObjectiveTo investigate the effects of Schisandrae Chinensis Fructus lignans on schizophrenia induced by dizocilpine maleate （MK-801） in mice and to clarify its mechanism. MethodsMale mice of 4-6 weeks old were randomized into blank， model， positive drug， and low-， medium-， and high-dose （40， 80， 160 mg·kg^-1， respectively） Schisandrae Chinensis Fructus lignans groups. The blank group was administrated with distilled water， and the other groups were injected with 0.5 mg·kg^-1 MK-801 to induce schizophrenia symptoms. Meanwhile， risperidone was injected at 0.2 mg·kg^-1 in the positive drug group， and mice in the intervention groups were injected with corresponding drugs for 14 consecutive days. The behavioral changes of mice were observed by autonomous activity test， open field test， forced swimming test， and water maze test. The levels of dopamine （DA） and 5-hydroxytryptamine （5-HT） in the brain and tumor necrosis factor-α （TNF-α） and nuclear factor-κB （NF-κB） in peripheral blood were quantified by enzyme-linked immunosorbent assay （ELISA）. The changes in the prefrontal lobe of mice were observed by hematoxylin-eosin staining， and the changes of the hippocampal tissue were observed by Nissl staining. The protein levels of silencing information regulatory factor 1 （SIRT1） and forkhead box protein O3a （FoxO3a） in the hippocampus of mice were determined by Western blot. ResultsCompared with the model group， low， medium， and high doses of Schisandrae Chinensis Fructus lignans reduced the total number of autonomous activities， total distance in the open field test， immobile time in the forced swimming test， and levels of TNF-α and NF-κB in peripheral blood （P<0.05）， while increasing the number of platform crossings in the water maze test and DA and 5-HT levels in the brain tissue （P<0.05）. Compared with the model group， risperidone and low， medium， and high doses of Schisandrae Chinensis Fructus lignans improve the neural cell morphology in the CA1 region， with full cells in neatly dense arrangement and exhibiting clear membrane boundary. Schisandrae Chinensis Fructus lignans inhibited the expression of SIRT 1 and FoxO3a in the hippocampus （P<0.05）. ConclusionTo sum up， Schisandrae Chinensis Fructus lignans may improve the behavior of schizophrenic mice by activating the SIRT1/FoxO3a signaling pathway to exert neuroprotective effects.

Objective: To investigate the prevalence and influencing factors of hepatitis C virus (HCV) infection among drug users in Baoshan District, Shanghai Municipality, so as to provide insights into strengthening HCV intervention among drug users. Methods: Drug users under community management in Baoshan District from 2017 to 2023 were recruited. Demographic information, drug use behaviors, sexual behaviors and receipt of intervention service were collected through questionnaire surveys. Blood samples were collected for HCV antibody testing, and the prevalence of anti-HCV antibody was analyzed. Factors affecting the prevalence of anti-HCV antibody among drug users were analyzed using a multivariable logistic regression model. Results: A total of 2 801 drug users were surveyed, including 2 233 males (79.72%) and 568 females (20.28%). The majority of drug users were aged 40 to <60 years (1 663 drug users, 59.37%). The prevalence of anti-HCV antibody was 28.35%, showing an overall upward trend from 2017 to 2023 (P<0.05). Multivariable logistic regression analysis showed that females (OR=1.468, 95%CI: 1.169-1.844), 40 years and over (40 to <50 years, OR=2.441, 95%CI: 1.838-3.242; 50 to <60 years, OR=2.377, 95%CI: 1.787-3.161; 60 to 97 years, OR=1.637, 95%CI: 1.163-2.304), using traditional drugs (OR=2.488, 95%CI: 1.967-3.147) or mixed drugs (OR=2.950, 95%CI: 1.974-4.409), having injected drugs (not share needles, OR=3.649, 95%CI: 2.849-4.673; share needles, OR=3.532, 95%CI: 1.851-6.738) and never using condoms during sexual contacts with spouses/cohabitants in the past year (OR=1.975, 95%CI: 1.354-2.879) were associated with a higher prevalence of anti-HCV antibody; the educational level of high school/technical secondary school (OR=0.483, 95%CI: 0.280-0.835) or college and above (OR=0.280, 95%CI: 0.129-0.608) was associated with a lower prevalence of anti-HCV antibody. Conclusions The prevalence of anti-HCV antibody among drug users in Baoshan District showed an upward trend from 2017 to 2023. Gender, age, educational level, type of drugs, history of drug injection and never using condoms during sexual contacts with spouses/cohabitants were influencing factors for prevalence of anti-HCV antibody among drug users.

Objective: To analyze the physical and chemical properties of human gastric factor by bioinformatics method , and to express and purify the protein. Methods: Protein online analysis software was used to predict the physical and chemical properties of gastric intrinsic factor and analyze their hydrophilicity and hydrophobicity. The online tool was used to predict and analyze the human gastric factor signal peptide and its subcellular location . The eukaryotic expression recombinant plasmid pcDNA3 . 1 ( + ) _human GIF_His tag was constructed , and the gastric intrinsic factor was purified by nickel column after expression in HEK293F cells . The purity and activity of purified gastric intrinsic factor were verified by SDS_PAGE , Western blot and indirect ELISA . Results: Gastric factor contained 417 amino acids and was a hydrophilic acid stable protein . It was a secreted protein with Sec original signal peptide . pcDNA3 . 1 ( + ) _human GIF_His tag recombinant plasmid was successfully constructed and soluble expression was obtained in HEK293F cell expression system. Conclusion The eukaryotic source of human gastric intrinsic factor is successfully prepared , and the bioinformatics results show that the protein is a hydrophilic acid stable secreted protein , laying a foundation for the subsequent use of this protein as an immunogen and protein calibrator to construct an immunoassay for gastric factor.

[Objective]To investigate the protective effect of Zhuangtongyin on the Middle Cerebral Artery Occlusion(MCAO)model by modulating ferroptosis through the Nrf2-SCL7A11/xCT-Gpx4 pathway and its underlying mechanism.[Methods]C57BL/6J mice were randomly divided into Sham operation group(Sham),model group(MCAO),low-dose Zhuangtongyin group(ZTY-L),high-dose Zhuangtongyin group(ZTY-H),with 5 mice in each group.The MCAO group was modelled by silica gel embolization,the middle cerebral artery of mice was embolized for 1h,then the silica gel was pulled out and reperfusion was performed after 72 h;and the other operations in the Sham group were the same as those in the MCAO group except that the thread plug was not inserted.The neural function of mice was evaluated by Zea-Longa method.TTC staining was used to evaluate the volume of cerebral infarction.The level of brain injury was evaluated by HE staining and Nissl staining.Prussian blue staining and the expression of iron transport-related carrier receptors TfR1 and DMT1 on mRNA level was detected by qPCR to evaluate the iron ion deposition level in mice brain.The expression of lipid peroxidation-related gene ACSL4 on mRNA level was detected by qPCR,and the content of 4-HNE was detected by ELISA kit to evaluate the lipid peroxidation level of mice brain.The expressions of ferroptosis marker PTGS2 mRNA level was detected by qPCR.The expressions of Nrf2,SCL7A11/xCT,Gpx4 in mice brain tissue were detected by Western-blot and immunofluorescence.[Results]Zhuangtongyin improved the nerve function of mice after MCAO(P＜0.05)and the cerebral infarction volume of mice(P＜0.05)and alleviate the pathological injury of cerebral cortex cells after MCAO operation.Zhuangtongyin attenuated the accumulation of trivalent iron ions in the brain tissue of mice following MCAO.Additionally,Zhuangtongyin downregulated the expression of TfR1 and DMT1 mRNA(P＜0.001),a transporter associated with cellular iron ion uptake,in the brains of post-MCAO mice.Furthermore,Zhuangtongyin reduced levels of lipid peroxidation product 4-HNE(P＜0.001)and suppressed ACSL4 mRNA expression in brain tissue post-MCAO(P＜0.001).Besides,Zhuangtongyin downregulated the expression of PTGS2 mRNA(P＜0.001),in the brains of post-MCAO mice.Zhuangtongyin increased the expression of nrf2 into the nucleus(P＜0.001),and increased the expression of xCT and Gpx4 in neurons after MCAO(P＜0.001).[Conclusion]Zhuangtongyin can enhance the nerve function and reduce cerebral infarction volume in MCAO/R mice,alleviate the pathological damage of cerebral cortex cells,and modulate the expression of key signaling molecules in the Nrf2-SCL7A11/xCT-Gpx4 pathway.Therefore,it is suggested that the mechanism by which Zhuangtongyin improves MCAO/R injury in mice may involve regulating ferroptosis through the Nrf2-SCL7A11/xCT-GPX4 pathway.

Objective To observe the efficacy of Zishen Ningxin capsule combined with telmisartan in the treatment of perimenopausal hypertension,and its influence on the patients'blood pressure,biochemical indexes and TCM symptom scores.Methods 300 patients with perimenopausal hypertension were randomly divided into control group and observation group with 150 patients in each group.The control group was given telmisartan treatment,and the observation group was given Zishen Ningxin capsule on the basis of the control group.The treatment course of both groups was 8 weeks.The dynamic blood pressure and coefficient of variation of blood pressure at 24 h before and after treatment were observed and compared between two groups(24 h systolic blood pressure and coefficient of variation,24 h diastolic blood pressure and its coefficient of variation),sex hormone levels(serum estradiol,follicle stimulating hormone,testosterone,progesterone),blood lipid levels(triacylglycerol,total cholesterol,low density lipoprotein cholesterol,high density lipoprotein cholesterol),renin and hypersensitive C-reactive protein levels,as well as the changes of TCM symptom score were evaluated for the efficacy of the two groups.Results After treatment,the 24 h systolic blood pressure and its coefficient of variation,24 h diastolic blood pressure and its coefficient of variation,the levels of sex hormones,blood lipids,renin and hypersensitive C-reactive protein,as well as TCM symptom scores in two groups were significantly improved(P＜0.05),and all indexes in the observation group were better than those in the control group(P＜0.05).The total effective rate of observation group was significantly higher than that of control group(P＜0.05).There was no significant difference of the incidence of adverse reactions in the two groups(P＞0.05).Conclusion Zishen Ningxin capsule combined with telmisartan has significant clinical efficacy in the treatment of perimenopausal hypertension,which can effectively reduce blood pressure,relieve symptoms,improve the levels of sex hormones,blood lipids,renin and hypersensitive C-reactive protein.

Objective To explore prokaryotic expression,purification of hemolysin coregulatory protein(Hcp)of Vibrio cholerae,and preparation of its polyclonal antibodies.Methods PCR was used to amplify Vibrio cholerae Hcp gene and clone it into pET28a vector to construct recombinant expression vector.The recombinant vector pET28a-hcp was transformed into E.coil BL21(DE3)for expression condition optimization and expression form identification.The soluble Hcp protein was purified by Ni-NTA column.The purified Hcp protein was used to immunize BALB/c mice to prepare polyclonal antibodies.The antibody titer was detected by indirect enzyme-linked immunosorbent assay(ELISA)to evaluate its immunogenicity.Western blot was used to analyze the specific recognition of antibodies to Hcp protein in Vibrio cholerae.Results The enzyme fragment digested by recombinant vector pET28a-hcp was consistent with the expected,the sequencing results were consistent with the Hcp gene sequence in the GenBank database,and the pET28a-hcp recombinant plasmid was successfully constructed.The recombinant plasmid was induced by isopropyl-β-D-thiogalactopyranoside(IPTG)to express the target protein with a relative molecular weight of 28 kD.The pure Hcp protein was obtained after purification by Ni-NTA column,and then Hcp polyclonal antibody(anti-Hcp)with a titer of 1∶512 000 could be obtained from immunized mice.Western blot results showed that anti-Hcp had specificity in recognizing Hcp protein in Vibrio cholerae.Conclusion The soluble expression of Hcp protein is successfully obtained,and high-titer polyclonal antibodies against Hcp are obtained after immunization of mice,which may lay a foundation for subsequent studies on the role of Hcp protein in the pathogenesis of T6SS in non-O1/non-O139 V.cholerae.

Dynamic tracking analysis of monoclonal antibodies (mAbs) biotransformation in vivo is crucial, as certain modifications could inactivate the protein and reduce drug efficacy. However, a particular challenge (i.e. immune recognition deficiencies) in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen. To address this limitation, a multi-epitope affinity technology utilizing the metal organic framework (MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region (CDR) mimotope peptide (HH24) and non-CDR mimotope aptamer (CH1S-6T) onto the surface of MOF@Au nanocomposite. Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited significantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope affinity technology. Moreover, the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HC-Asn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry (LC-QTOF-MS). Therefore, multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb. Particularly, the novel bioanalytical platform can be successfully used for the tracking analysis of trastuzumab modifications in different biological fluids. Compared to the spiked phosphate buffer (PB) model, faster modification trends were monitored in the spiked serum and patients' sera due to the catalytic effect of plasma proteins and relevant proteases. Differences in peptide modification levels of trastuzumab in patients' sera were also monitored. In summary, the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb, contributing to improved understanding and paving the way for future research and clinical applications.

Dynamic tracking analysis of monoclonal antibodies(mAbs)biotransformation in vivo is crucial,as certain modifications could inactivate the protein and reduce drug efficacy.However,a particular chal-lenge(i.e.immune recognition deficiencies)in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen.To address this limitation,a multi-epitope affinity technology utilizing the metal organic framework(MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region(CDR)mimotope peptide(HH24)and non-CDR mimotope aptamer(CH1S-6T)onto the surface of MOF@Au nanocomposite.Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited signifi-cantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope af-finity technology.Moreover,the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HC-Asn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry(LC-QTOF-MS).Therefore,multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb.Particularly,the novel bioanalytical platform can be suc-cessfully used for the tracking analysis of trastuzumab modifications in different biological fluids.Compared to the spiked phosphate buffer(PB)model,faster modification trends were monitored in the spiked serum and patients'sera due to the catalytic effect of plasma proteins and relevant proteases.Differences in peptide modification levels of trastuzumab in patients'sera were also monitored.In summary,the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb,contributing to improved understanding and paving the way for future research and clinical applications.

OBJECTIVE: To establish the mode of anterior cervical surgery in outpatient setting, and evaluate its preliminary effectiveness. METHODS: A clinical data of patients who underwent anterior cervical surgery between January 2022 and September 2022 and met the selection criteria was retrospectively analyzed. The surgeries were performed in outpatient setting ( n=35, outpatient setting group) or in inpatient setting ( n=35, inpatient setting group). There was no significant difference between the two groups ( P>0.05) in age, gender, body mass index, smoking, history of alcohol drinking, disease type, the number of surgical levels, operation mode, as well as preoperative Japanese Orthopaedic Association (JOA) score, visual analogue scale score of neck pain (VAS-neck), and visual analogue scale score of upper limb pain (VAS-arm). The operation time, intraoperative blood loss, total hospital stay, postoperative hospital stay, and hospital expenses of the two groups were recorded; JOA score, VAS-neck score, and VAS-arm score were recorded before and immediately after operation, and the differences of the above indexes between pre- and post-operation were calculated. Before discharge, the patient was asked to score satisfaction with a score of 1-10. RESULTS: The total hospital stay, postoperative hospital stay, and hospital expenses were significantly lower in the outpatient setting group than in the inpatient setting group ( P<0.05). The satisfaction of patients was significantly higher in the outpatient setting group than in the inpatient setting group ( P<0.05). There was no significant difference between the two groups in operation time and intraoperative blood loss ( P>0.05). The JOA score, VAS-neck score, and VAS-arm score of the two groups significantly improved at immediate after operation when compared with those before operation ( P<0.05). There was no significant difference in the improvement of the above scores between the two groups ( P>0.05). The patients were followed up (6.67±1.04) months in the outpatient setting group and (5.95±1.90) months in the inpatient setting group, with no significant difference ( t=0.089, P=0.929). No surgical complications, such as delayed hematoma, delayed infection, delayed neurological damage, and esophageal fistula, occurred in the two groups. CONCLUSION The safety and efficiency of anterior cervical surgery performed in outpatient setting were comparable to that performed in inpatient setting. Outpatient surgery mode can significantly shorten the postoperative hospital stay, reduce hospital expenses, and improve the patients' medical experience. The key points of the outpatient mode of anterior cervical surgery are minimizing damage, complete hemostasis, no drainage placement, and fine perioperative management.
Humans ; Treatment Outcome ; Cervical Vertebrae/surgery* ; Outpatients ; Retrospective Studies ; Blood Loss, Surgical ; Spinal Fusion ; Neck Pain

Cluster of differentiation 36 (CD36) is a membrane glycoprotein receptor capable of binding and transporting fatty acid. Nogo-B regulates the metabolism of fatty acids in the liver and affects the development of liver cancer. To date, it remains unclear whether the interaction between CD36 and Nogo-B affects the proliferation and migration of breast cancer cells. In the current study, we aimed to determine whether the interference of CD36 and Nogo-B affects the proliferation and migration of triple-negative breast cancer (TNBC) cells. The results showed that inhibition of CD36 or Nogo-B alone can inhibit the proliferation and migration of TNBC cells, and the inhibitory effect was more pronounced when CD36 and Nogo-B were inhibited simultaneously. Meanwhile, it was found that inhibition of CD36 and Nogo-B expression can inhibit the expression of Vimentin, B-cell lympoma-2 (BCL2) and proliferating cell nuclear antigen (PCNA). In vivo, knockdown of CD36 or Nogo-B in E0771 cells reduced its tumorigenic ability, which was further enhanced by knockdown of CD36 and Nogo-B simultaneously. Mechanistically, inhibition of CD36 and Nogo-B expression can decrease fatty acid binding protein 4 (FABP4) and fatty acid transport protein 4 (FATP4) expression. Moreover, overexpression of CD36 and Nogo-B-induced cell proliferation was attenuated by FABP4 siRNA, indicating that inhibition of CD36 and Nogo-B expression could inhibit the absorption and transport of fatty acids, thereby inhibiting the proliferation and migration of TNBC. Furthermore, inhibition of CD36 and Nogo-B expression activated the P53-P21-Rb signaling pathway which contributed to the CD36 and Nogo-B-inhibited proliferation and migration of TNBC. Taken together, the results suggest that inhibition of CD36 and Nogo-B can reduce the proliferation and migration of TNBC, which provides new targets for the development of drugs against TNBC.
Humans ; Triple Negative Breast Neoplasms/metabolism* ; Cell Movement ; Cell Proliferation ; Cell Line, Tumor ; Fatty Acids