Objective To investigate the mechanism of action and potential biomarkers of Jiangzhi Qingshen capsules in the treatment of dyslipidaemia based on clinical lipid metabolomics.Methods 30 patients with dyslipidaemia from Zhengzhou Hospital of Chinese Medicine were collected as the test group,and their lipid levels before and after taking Jiang Zhi Qing Shen capsule for 12 weeks were compared.Another 30 healthy patients were enrolled in the physical examination department of Zhengzhou Hospital of Chinese Medicine,and metabolomics investigation was carried out on plasma samples of the test group before and after the treatment as well as those of the healthy patients by LC-MS/MS technology.The differences between the groups were compared by multivariate statistical analysis,and the potential biomarkers were identified by HMDB and lipidblast databases,so as to clarify the possible pathways and targets for the treatment of mild dyslipidaemia by Jiangzhi Qingshen capsules.Results Jiangzhi Qingshen capsules could improve patients' blood lipid level,BMI and abdominal circumference significantly(P<0.05).And metabolomics results showed that 29 lipid metabolites in the plasma of the treated patients were dialled back,which involved cholesterol metabolism,fat digestion and absorption,glycerol-phospholipid metabolism and other biological processes.Conclusion The efficacy of Jiangzhi Qingshen capsules in treating dyslipidaemia was confirmed,and its mechanism of action might be related to the regulation of lipid metabolism.Metabolites such as acylcarnitine(Acar),phosphatidylethanolamine(PE)and phosphatidylcholine(PC)were expected to be used as the biomarkers of dyslipidaemia,so as to provide objective scientific basis for the lipid-lowering capsule,and to facilitate the promotion of its application.

Objective To investigate the clinical efficacy of the traditional Chinese medicine(TCM)throat opening and brightening method in treating unilateral vocal cord paralysis(UVCP).Methods Sixty patients with UVCP were prospectively collected and randomly assigned to two groups:the Chinese herbal medicine group(trea-ted with Buyang Huanwu Decoction,n=30)and the throat opening and brightening method group(treated with TCM throat opening and brightening method,n=30).The clinical studies that had utilized injection laryngoplasty for the treatment of UVCP(historical control group).Evaluation indicators included the voice handicap index-10(VHI-10),GRBAS-G,objective acoustic measurements of voice(vocal intensity,F0,shimmer,jitter,HNR),and aerodynamic measurements(maximum phonation time,MPT).Results Before treatment,no significant differences were observed between the two groups in all the evaluation indicators(P＞0.05).Post-treatment,the throat open-ing and brightening method group demonstrated significant improvements in VHI-10,GRBAS-G,shimmer,jitter,HNR,and MPT compared to pre-treatment values(P＜0.01),and these improvements were superior to those in the Chinese herbal medicine group.Pre-treatment VHI-10,GRBAS-G,and shimmer scores in the throat opening and brightening method group were significantly higher than those in the historical literature group(P＜0.01).Af-ter treatment,no significant differences were noted in any assessed parameters between the two groups(P＞0.05).Conclusion The TCM throat opening and brightening method significantly enhances phonatory function and quality of life in patients with UVCP,showing comparable efficacy to injection laryngoplasty.

A long-acting feline ω-interferon fusion protein(FSA-FeIFNω)was designed and its bio-logical function validated.According to the optimization of the sequence of feline serum albumin and feline ω interferon in NCBI,the recombinant plasmid pET-30a(+)-FSA-FeIFNω was con-structed,which was transformed into E.coli BL21(DE3)competent cells,the expression of re-combinant protein FSA-FeIFNω was induced by IPTG,and the expressed inclusion body protein was identified by Western blot,the refolding product was purified by Ni-NTA affinity chromatog-raphy,and the concentration of dialysis and concentrated protein after purification was determined by BCA method.The antiviral activity of recombinant protein was detected by micro-cytopathic in-hibition method in the CRFK/VSV system,the in vitro half-life was detected by 50％mouse plas-ma method,the tumor cell proliferation inhibition activity was detected by MTT method,and anti-tumor activity was detected by mouse melanoma model.The pET-30a(+)-FeIFNω and pET-30a(+)-FSA-FeIFNω expression vectors were successfully constructed,and 87 kDa recombinant FSA-FeIFNω protein was obtained in E.coli,with a purified protein purity of 95％,with a concen-tration of 1 g/L,and the biological activity was 2.56 × 106 IU/mg,the plasma half-life was significantly prolonged(＞24 h),and the half-inhibitory concentration IC50 of B16-F10 in mouse melanoma cells was 56.01 mg/L.The FSA-FeIFNω group significantly inhibited tumor growth,and the treatment effect was better than that of the control group and other experimental groups.The recombinant FSA-FeIFNω protein obtained in this study had long-acting effect and good biological activity.

To develop a novel immunocastration vaccine for animals,researchers designed and syn-thesized the recombinant plasmid pET-30a-SF which could express the recombinant protein SF.This protein was then conjugated in vitro with the synthetic peptide STGP to prepare the SF-STGP nanoparticle vaccine,and its immunocastration effect on mice was studied.The Spy Catcher and ferritin amino acid sequences were connected via GGGGS,and after codon optimization for E.coli,the recombinant plasmid pET-30a-SF was constructed and transformed into E.coli for in-duced expression.The recombinant protein SF was purified using Ni-column affinity chromatogra-phy and characterized.The peptide STGP,composed of Spy Tag,GnRH,and PADRE connected by GGGS,was conjugated with the recombinant protein SF in vitro.The self-assembled nanoparticles were observed using transmission electron microscopy(TEM)and dynamic light scattering(DLS).The prepared SF-STGP nanoparticles were mixed with MONTANIDE ISA 206 VG at a 1∶1 ratio to form the vaccine,which was then subcutaneously injected into male BALB/c mice for immunocastration evaluation.The recombinant protein SF showed the highest soluble expression when induced at 18 ℃ with 0.25 mmol/L IPTG for 14 h,and the maximum conjugation efficiency with STGP was achieved at a 1∶8 molar ratio.TEM and DLS analyses revealed that both the re-combinant protein SF and SF-STGP could self-assemble into nanoparticles with average diameters of 16.2 nm and 17.8 nm,respectively.Mouse immunization results demonstrated that the SF-STGP nanoparticle vaccine generated specific GnRH antibodies after the first immunization,with the spe-cific antibody D45o reaching its peak at the 10 th week.The SF-STGP+ISA 206 immunization group showed a peak D450 value of 2.8,and the specific antibody levels in all immunization groups were significantly higher than those in the control group(P＜0.05).Additionally,the SF-STGP nanoparticle vaccine effectively reduced serum testosterone levels in mice,with the testosterone concentration in the immunization groups being significantly lower than that in the control group(P＜0.05).Compared to the control group,the immunization group exhibits testicular atrophy.The constructed SF-STGP nanoparticle vaccine proves to be a highly effective immunogen,capable of inducing testicular atrophy and reducing gonadal hormone concentrations,demonstrating excellent castration effects.This study provides new insights into immunocastration vaccines for mammals.

Objective To investigate the mechanism of action and potential biomarkers of Jiangzhi Qingshen capsules in the treatment of dyslipidaemia based on clinical lipid metabolomics.Methods 30 patients with dyslipidaemia from Zhengzhou Hospital of Chinese Medicine were collected as the test group,and their lipid levels before and after taking Jiang Zhi Qing Shen capsule for 12 weeks were compared.Another 30 healthy patients were enrolled in the physical examination department of Zhengzhou Hospital of Chinese Medicine,and metabolomics investigation was carried out on plasma samples of the test group before and after the treatment as well as those of the healthy patients by LC-MS/MS technology.The differences between the groups were compared by multivariate statistical analysis,and the potential biomarkers were identified by HMDB and lipidblast databases,so as to clarify the possible pathways and targets for the treatment of mild dyslipidaemia by Jiangzhi Qingshen capsules.Results Jiangzhi Qingshen capsules could improve patients' blood lipid level,BMI and abdominal circumference significantly(P<0.05).And metabolomics results showed that 29 lipid metabolites in the plasma of the treated patients were dialled back,which involved cholesterol metabolism,fat digestion and absorption,glycerol-phospholipid metabolism and other biological processes.Conclusion The efficacy of Jiangzhi Qingshen capsules in treating dyslipidaemia was confirmed,and its mechanism of action might be related to the regulation of lipid metabolism.Metabolites such as acylcarnitine(Acar),phosphatidylethanolamine(PE)and phosphatidylcholine(PC)were expected to be used as the biomarkers of dyslipidaemia,so as to provide objective scientific basis for the lipid-lowering capsule,and to facilitate the promotion of its application.

Objective To investigate the clinical efficacy of the traditional Chinese medicine(TCM)throat opening and brightening method in treating unilateral vocal cord paralysis(UVCP).Methods Sixty patients with UVCP were prospectively collected and randomly assigned to two groups:the Chinese herbal medicine group(trea-ted with Buyang Huanwu Decoction,n=30)and the throat opening and brightening method group(treated with TCM throat opening and brightening method,n=30).The clinical studies that had utilized injection laryngoplasty for the treatment of UVCP(historical control group).Evaluation indicators included the voice handicap index-10(VHI-10),GRBAS-G,objective acoustic measurements of voice(vocal intensity,F0,shimmer,jitter,HNR),and aerodynamic measurements(maximum phonation time,MPT).Results Before treatment,no significant differences were observed between the two groups in all the evaluation indicators(P＞0.05).Post-treatment,the throat open-ing and brightening method group demonstrated significant improvements in VHI-10,GRBAS-G,shimmer,jitter,HNR,and MPT compared to pre-treatment values(P＜0.01),and these improvements were superior to those in the Chinese herbal medicine group.Pre-treatment VHI-10,GRBAS-G,and shimmer scores in the throat opening and brightening method group were significantly higher than those in the historical literature group(P＜0.01).Af-ter treatment,no significant differences were noted in any assessed parameters between the two groups(P＞0.05).Conclusion The TCM throat opening and brightening method significantly enhances phonatory function and quality of life in patients with UVCP,showing comparable efficacy to injection laryngoplasty.

A long-acting feline ω-interferon fusion protein(FSA-FeIFNω)was designed and its bio-logical function validated.According to the optimization of the sequence of feline serum albumin and feline ω interferon in NCBI,the recombinant plasmid pET-30a(+)-FSA-FeIFNω was con-structed,which was transformed into E.coli BL21(DE3)competent cells,the expression of re-combinant protein FSA-FeIFNω was induced by IPTG,and the expressed inclusion body protein was identified by Western blot,the refolding product was purified by Ni-NTA affinity chromatog-raphy,and the concentration of dialysis and concentrated protein after purification was determined by BCA method.The antiviral activity of recombinant protein was detected by micro-cytopathic in-hibition method in the CRFK/VSV system,the in vitro half-life was detected by 50％mouse plas-ma method,the tumor cell proliferation inhibition activity was detected by MTT method,and anti-tumor activity was detected by mouse melanoma model.The pET-30a(+)-FeIFNω and pET-30a(+)-FSA-FeIFNω expression vectors were successfully constructed,and 87 kDa recombinant FSA-FeIFNω protein was obtained in E.coli,with a purified protein purity of 95％,with a concen-tration of 1 g/L,and the biological activity was 2.56 × 106 IU/mg,the plasma half-life was significantly prolonged(＞24 h),and the half-inhibitory concentration IC50 of B16-F10 in mouse melanoma cells was 56.01 mg/L.The FSA-FeIFNω group significantly inhibited tumor growth,and the treatment effect was better than that of the control group and other experimental groups.The recombinant FSA-FeIFNω protein obtained in this study had long-acting effect and good biological activity.

To develop a novel immunocastration vaccine for animals,researchers designed and syn-thesized the recombinant plasmid pET-30a-SF which could express the recombinant protein SF.This protein was then conjugated in vitro with the synthetic peptide STGP to prepare the SF-STGP nanoparticle vaccine,and its immunocastration effect on mice was studied.The Spy Catcher and ferritin amino acid sequences were connected via GGGGS,and after codon optimization for E.coli,the recombinant plasmid pET-30a-SF was constructed and transformed into E.coli for in-duced expression.The recombinant protein SF was purified using Ni-column affinity chromatogra-phy and characterized.The peptide STGP,composed of Spy Tag,GnRH,and PADRE connected by GGGS,was conjugated with the recombinant protein SF in vitro.The self-assembled nanoparticles were observed using transmission electron microscopy(TEM)and dynamic light scattering(DLS).The prepared SF-STGP nanoparticles were mixed with MONTANIDE ISA 206 VG at a 1∶1 ratio to form the vaccine,which was then subcutaneously injected into male BALB/c mice for immunocastration evaluation.The recombinant protein SF showed the highest soluble expression when induced at 18 ℃ with 0.25 mmol/L IPTG for 14 h,and the maximum conjugation efficiency with STGP was achieved at a 1∶8 molar ratio.TEM and DLS analyses revealed that both the re-combinant protein SF and SF-STGP could self-assemble into nanoparticles with average diameters of 16.2 nm and 17.8 nm,respectively.Mouse immunization results demonstrated that the SF-STGP nanoparticle vaccine generated specific GnRH antibodies after the first immunization,with the spe-cific antibody D45o reaching its peak at the 10 th week.The SF-STGP+ISA 206 immunization group showed a peak D450 value of 2.8,and the specific antibody levels in all immunization groups were significantly higher than those in the control group(P＜0.05).Additionally,the SF-STGP nanoparticle vaccine effectively reduced serum testosterone levels in mice,with the testosterone concentration in the immunization groups being significantly lower than that in the control group(P＜0.05).Compared to the control group,the immunization group exhibits testicular atrophy.The constructed SF-STGP nanoparticle vaccine proves to be a highly effective immunogen,capable of inducing testicular atrophy and reducing gonadal hormone concentrations,demonstrating excellent castration effects.This study provides new insights into immunocastration vaccines for mammals.

Objective To analyze risk factors of acute respiratory failure(ARF)in patients with hypertriglyceridemia acute pancreatitis(HTG-AP)and construct a risk prediction model.Methods A total of 222 HTG-AP patients were included in this study and divided into the non-ARF group(176 cases)and the ARF group(46 cases)according to diagnostic guidelines for ARF.Clinical data of the two groups were compared and the predictive factors were screened.These selected factors were then utilized in a multivariate Logistic regression analysis to construct a Logistic regression model.Subsequent evaluation of the model′s predictive ability,accuracy and clinical utility was conducted through ROC,curve analysis,calibration plot examination and decision curve analysis(DCA),respectively.Results Compared with the non-ARF group,the levels of high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL)-C and albumin(ALB)were decreased in the ARF group(P＜0.05),while the levels of creatinine(Cr),urea nitrogen(BUN),aspartate aminotransferase(AST)and C-reactive protein(CRP)were increased,and the incidence of pleural fluid and ascites was also increased(P＜0.05).Multivariate Logistic regression analysis showed that higher levels of Cr and AST,lower levels of ALB,HDL-C and ascites were independent risk factors for HTG-AP complicated ARF(P＜0.05).Based on these results,a column-line prediction model for HTG-AP complicated ARF was established.After internal verification,the area under curve(AUC)of receiver operating characteristic(ROC)curve of the nomogram model was 0.952(95%CI:0.923-0.981),the Youden index was 0.808 and the sensitivity and specificity were 93.33%and 87.43%,respectively.The calibration curve showed that the probability of HTG-AP concurrent ARF predicted by the model was in good agreement with the actual probability.The DCA curve showed that the model had certain clinical value.Conclusion The nomogram prediction model combined could provide a scheme for the clinical prevention of HTG-AP complicated with ARF.

Objective:To observe the effect of pecking moxibustion on the pattern characteristics of redness,swelling,heat,and pain in the affected joints,also the synovial cell ultrastructure in rats with rheumatoid arthritis(RA)due to damp heat affecting bones/joints,and to explore the anti-inflammatory mechanism of pecking moxibustion in treating the early stage of RA. Methods:Eighteen rats were randomly selected from 78 female ones as the blank group,and all the other rats were subjected to preparing the"differentiation of disease and pattern"RA model due to damp heat affecting bones/joints by using the method of"collagen-induced arthritis plus windy,damp,and hot environment stimulation".Fifty-four rats with successful modeling were randomly divided into a model group,a drug group,and a pecking moxibustion group,with 18 rats in each group.Rats in the drug group were given methotrexate at a dose of 1 mg/(kg·bw)on the 1st,8th,and 15th days.Rats in the pecking moxibustion group were treated with pecking moxibustion at Quchi(LI11),Dazhui(GV14),and Ashi points,and each point was treated with moxibustion for 15 min every day and a total of 3 courses of treatment,with 6 d as a course of treatment.After treatment,the capillary permeability,joint swelling,joint surface temperature,and plantar thermal pain threshold of the diseased joints in rats were observed,and the ultrastructural changes of synovial cells were observed by transmission electron microscopy. Results:The local swelling,surface temperature,and Evans blue(EB)leakage volume were significantly higher(P<0.05),the thermal pain threshold was significantly lower(P<0.05),and the synovial cell ultrastructure was obviously damaged in the affected joints in the model group compared with the blank group.The swelling degree,surface temperature,and EB leakage volume were significantly reduced(P<0.05),the thermal pain threshold was significantly increased(P<0.05),and the ultrastructural abnormalities of synovial cells were significantly improved in the diseased joints in the drug group and the pecking moxibustion group compared with the model group.The thermal pain threshold of rats in the pecking moxibustion group was significantly improved compared with the drug group(P<0.05). Conclusion:Pecking moxibustion obviously improves the pattern characteristics of local redness,swelling,heat,and pain in the diseased joints of rats with RA due to damp heat affecting bones/joints and effectively repairs the ultrastructure of the damaged synovium.It suggests that the pecking moxibustion intervention has a significant anti-inflammatory effect on early RA.