1.Clinical Observation on Modified Shehuang Ointment for the Treatment of Facial Seborrheic Dermatitis
Man-Xin HUANG ; De-Jian DUAN ; Hai-Fang GAN ; Dan HUANG ; Jia-Yao NIE
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(1):110-115
		                        		
		                        			
		                        			Objective To observe the clinical efficacy of modified Shehuang Ointment(mainly composed of Cnidii Fructus,Phellodendri Chinensis Cortex,and Zanthoxyli Pericarpium)for the treatment of facial seborrheic dermatitis(SD).Methods Seventy-two patients with facial SD were randomly divided into observation group and control group,with 36 patients in each group.Both groups of patients were given oral use of Acrivastine Capsules and Vitamin B6 Tablets,and additionally,the observation group was given topical application of modified Shehuang Ointment and the control group was given topical application of 2%Ketoconazole cream.The course of treatment covered 4 weeks.The changes of clinical symptom scores and dermatology life quality index(DLQI)scores in the two groups were observed before and after treatment,and the clinical efficacy and safety of the two groups were also evaluated.Results(1)After 4 weeks of treatment,the total effective rate of the observation group was 88.89%(32/36),and that of the control group was 72.22%(26/36).The intergroup comparison showed that the efficacy of the observation group was significantly superior to that of the control group,and the difference was statistically significant(P<0.05).(2)After treatment,the clinical symptom scores of erythema,scales,grease,rash area,itchiness and other clinical symptoms of the patients in the two groups were significantly decreased compared with those before treatment(P<0.05),and the clinical symptom scores of the observation group were significantly lower than those of the control group,the differences being statistically significant(P<0.05).(3)After treatment,the DLQI scores of patients in the two groups were significantly lower than those before treatment(P<0.05),and the DLQI scores in the observation group were significantly lower than those in the control group after treatment,the difference being statistically significant(P<0.05).(4)During the treatment period,no significant adverse reactions occurred in the two groups of patients,with high safety.Conclusion The conventional western medicine treatment combined with topical application of modified Shehuang Ointment exerts certain effect in the treatment of facial SD,which can effectively relieve the clinical symptoms and improve the quality of life of patients.
		                        		
		                        		
		                        		
		                        	
2.Screening and identification of human monoclonal antibodies against low-calcium response V antigen of Yersinia pestis
Li ZHANG ; Bin-Yang ZHENG ; Qi ZHANG ; Hai-Lian WU ; Hong-Xin PAN ; Feng-Cai ZHU ; Hai-Sheng WU ; Jian-Fang ZHOU
Chinese Journal of Zoonoses 2024;40(1):15-20
		                        		
		                        			
		                        			To characterize human antibodies against low-calcium response V(LcrV)antigen of Yersinia pestis,the mono-clonal antibodies were screened and assayed.Antibody gene was derived from peripheral blood mononuclear cells of the vaccin-ees immunized by plague subunit vaccine in phase Ⅱb clinical trial.Human ScFv antibody library was constructed by phage dis-play.After panning library by using recombinant LcrV antigen,antibody variable genes were sequenced and converted into IgG1 format to evaluate its binding specificity and relevant parameters.An anti-plague human ScFv antibody library was estab-lished contained 7.54× 108 independent clones.After panning by LcrV antigen,3 human antibodies named as RV-B4,RV-D1 and RV-E8,respectively,were identified.Using indirect enzyme-linked immunosorbent assay(ELISA)and Western blot(WB),the specific bindings of the mAbs to LcrV antigen were confirmed.The dissociation constant(KD)of them to LcrV is 2.1 nmol/L,1.24 nmol/L and 42 nmol/L,respectively.Minor protective efficacy was found among 3 human antibodies in Y.pestis 141-infected mice.Three anti-LcrV monoclonal antibodies generated from immunized vaccinees were binding specific antibod-ies and could not block plague infection in mice.These antibodies are the potential candidate reagents for basic research of plague immunity and the application of plague diagnosis.
		                        		
		                        		
		                        		
		                        	
3.Establishment and application of a prospective follow-up research method for acute infectious diseases in Shanghai community residents
Yaxu ZHENG ; Xiao YU ; Huanyu WU ; Liming WU ; Jian CHEN ; Wenjia XIAO ; Zhuoying HUANG ; Sheng LIN ; Qiwen FANG ; Rui LIU ; Hao ZHANG ; Xin CHEN
Shanghai Journal of Preventive Medicine 2024;36(1):5-10
		                        		
		                        			
		                        			ObjectiveTo present the exploration and application of a prospective follow-up research method for acute infectious disease surveillance based on natural community populations, using COVID-19 infection as an example, and to provide a reference for improving the infectious disease surveillance and early warning system. MethodsA multi-stage probability proportional sampling method was employed to sample residents from all communities of 16 administrative districts in Shanghai, with households as the units. A cohort for acute infectious diseases based on natural community populations was established. The baseline survey was conducted for all cohort subjects, and COVID-19 antigen test kits were distributed. From December 21, 2022 to September 30, 2023, prospective follow-up monitoring of COVID-19 antigen and nucleic acid was carried out on the study subjects on a weekly basis. The baseline characteristics and follow-up information of the cohort subjects were described. ResultsThe cohort for acute infectious diseases included a total of 12 881 subjects, comprising 6 098 males (47.3%) and 6 783 females (52.7%). The baseline survey revealed that 35.2% (4 540/12 881) of the subjects had a history of COVID-19 infection. During the follow-up period from December 21, 2022 to September 30, 2023, the average incidence density in the cohort was 0.61/person-year, with a higher incidence density in females (0.63/person-year) compared to males (0.59/person-year). Individuals aged 60 and above (0.64/person-year) and those with underlying health conditions (0.67/person-year) had a higher incidence density. Healthcare workers showed a notably higher incidence density (0.84/person-year) than that in other occupational groups. As of September 30, 2023, a total of 340 subjects in the cohort experienced secondary infections, with a median interval of 170 days between the first and second infections. ConclusionThis study applies cohort study method to acute infectious disease surveillance, providing crucial data support for estimating infection rates and forecasting alerts for acute infectious diseases in the community. This method can be promoted and applied as a new approach for acute infectious disease surveillance. 
		                        		
		                        		
		                        		
		                        	
4.Total saponins from Trillium tschonoskii maxim alleviates cerebral ischemia-reperfusion injury by inhibiting ferroptosis through Keap-1/Nrf2/HO-1 and Nrf2/SLC7A11/GPX4 pathways
Jian-Hong GAO ; Tian-Ying SONG ; Chao-Xi TIAN ; Fang-Yu ZHAO ; Yi-Duo HE ; Xin LIU ; Xian-Bing CHEN
Chinese Pharmacological Bulletin 2024;40(10):1850-1857
		                        		
		                        			
		                        			Aim To examine the neuroprotective im-pacts of total saponins from Trillium tschonoskii maxim(TST)on cerebral ischemia-reperfusion injury(CIRI)in rats and delve into the mechanisms of ferroptosis.Methods The CIRI model was prepared by dividing male SD rats into the model group,TST(0.1 g·kg-1)group,Donepezil hydrochloride(0.45 mg·kg-1)group,and sham group.The cognitive functions of rats in each group were assessed through the Morris water maze test,the changes in neurological function were evaluated using the Zea-Longa method,the infarct area was observed via TTC staining,and the pathologi-cal alterations in brain tissue were analysed using HE and Nissl staining.To further investigate the underly-ing mechanism,the mitochondrial structural changes were examined using transmission electron microscopy,and the levels of GSH-PX,MDA,and SOD were ana-lyzed.Additionally,the expressions of GPX4 and Nrf2 proteins were evaluated through immunohistochemistry and immunofluorescence.Furthermore,the protein lev-els of Keap1/Nrf2/HO-1 and Nrf2/SLC7A11/GPX4 pathways in rats were examined using Western blot-ting.Results The rats in the model group displayed diminished learning and memory capabilities in com-parison to those in the sham group,as well as a signifi-cantly increased cerebral infarction area and higher neurological function scores(P<0.01),significantly increased cerebral infarct area,disordered and loosely arranged neurons,and reduced Nissl bodies.Addition-ally,mitochondria showed typical signs of ferroptosis.Changes related to ferroptosis included decreased activ-ities of SOD and GSH-PX(P<0.01)and increased MDA levels(P<0.01).The expression of GPX4 and Nrf2-positive cells was significantly reduced,along with decreased fluorescence intensity of GPX4.Further-more,the protein expression of Keap1,Nrf2,HO-1,GPX4,SLC7A11 in the hippocampus decreased(P<0.05,P<0.01).Following the administration of TST,these effects showed improvement.Conclusions TST has neuroprotective effects,enhancing learning and memory abilities while reducing oxidative stress levels.The mechanism may involve the inhibition of ferroptosis through the Keap-1/Nrf2/HO-1 and Nrf2/SLC7 A11/GPX4 pathways.
		                        		
		                        		
		                        		
		                        	
5.mfat-1 gene therapy prevents and ameliorates multiple sclerosis in mice
Min-Yi TANG ; Xin-Yun BI ; Shuai WANG ; Chao-Feng XING ; Xiao-Li WU ; Zi-Jian ZHAO ; Fang-Hong LI
Chinese Pharmacological Bulletin 2024;40(10):1930-1936
		                        		
		                        			
		                        			Aim To investigate the preventive and therapeutic effects of the mfat-1 gene therapy on exper-imental autoimmune encephalomyelitis in mice.Meth-ods mfat-1 gene therapy was used to render the host endogenous capability of producing ω-3 PUFAs,con-comitantly reduce the levels of ω-6 PUFAs,and change the proportion of ω-3/ω-6 PUFAs.Then,the levels of PUFAs in blood were analyzed by gas chromatography.The neurological deficits in mice were evaluated by neurological dysfunction score.HE staining and LFB staining of mouse spinal cord slices were used to ob-serve central nervous system inflammation infiltration and demyelinating lesions.Flow cytometry microsphere microarray technology was used to detect the content of cytokines in serum.Results The mfat-1 gene therapy could significantly raise the proportion of ω-3/ω-6 PU-FAs(P<0.05),markedly delay the incubation period and peak period and reduce neurological dysfunction scores(P<0.05),and improve inflammation and de-myelination of spinal cords(P<0.05).It could also greatly increase the levels of IL-2,IFN-γ,IL-4 and IL-17 in serum(P<0.05).Conclusion The pro-portion of ω-3/ω-6 PUFAs in blood circulation en-hanced by mfat-1 gene therapy can effectively prevent and treat experimental autoimmune encephalomyelitis in mice.
		                        		
		                        		
		                        		
		                        	
6.Radix Angelica Sinensis and Radix Astragalus ultrafiltration extract improves radiation-induced pulmonary fibrosis in rats by regulating NLRP3/caspase-1/GSDMD pyroptosis pathway
Chun-Zhen REN ; Jian-Fang YUAN ; Chun-Ling WANG ; Xiao-Dong ZHI ; Qi-Li ZHANG ; Qi-Lin CHEN ; Xin-Fang LYU ; Xiang GAO ; Xue WU ; Xin-Ke ZHAO ; Ying-Dong LI
Chinese Pharmacological Bulletin 2024;40(11):2124-2131
		                        		
		                        			
		                        			Aim To investigate the mechanism of py-roptosis mediated by the NLRP3/caspase-1/GSDMD signaling pathway and the intervention effect of Radix Angelica Sinensis and Radix Astragalus ultrafiltration extract(RAS-RA)in radiation-induced pulmonary fi-brosis.Methods Fifty Wistar rats were randomly di-vided into five groups,with ten rats in each group.Ex-cept for the blank control group,all other groups of rats were anesthetized and received a single dose of 40 Gy X-ray local chest radiation to establish a radiation-in-duced pulmonary fibrosis rat model.After radiation,the rats in the RAS-RA intervention groups were orally administered doses of 0.12,0.24 and 0.48 g·kg-1 once a day for 30 days.The average weight and lung index of the rats were observed after 30 days of contin-uous administration.Hydroxyproline(HYP)content in lung tissue was determined by hydrolysis method.The levels of IL-18 and IL-1 β in serum were detected by ELISA.Lung tissue pathological changes were ob-served by HE and Masson staining.Ultrastructural changes in lung tissue were observed by transmission e-lectron microscopy.The expression levels of NLRP3/caspase-1/GSDMD pyroptosis pathway-related proteins and fibrosis-related proteins in lung tissue were detec-ted by Western blot.Results Compared with the blank group,the HYP content in lung tissue and the levels of IL-18 and IL-1 β in serum significantly in-creased in the model group(P<0.01).HE and Mas-son staining showed inflammatory cell infiltration and collagen fiber deposition.Transmission electron mi-croscopy revealed increased damaged mitochondria,disordered arrangement,irregular morphology,shallow matrix,outer membrane rupture,mostly fractured and shortened cristae,mild expansion,increased electron density of individual mitochondrial matrix,mild sparse structure of lamellar bodies,partial disorder,unclear organelles,and characteristic changes of pyroptosis.Western blot analysis showed increased expression of caspase-1,GSDMD,NLRP3,CoL-Ⅰ,α-SMA,and CoL-Ⅲ proteins(P<0.01).Compared with the model group,the RAS-RA intervention group showed signifi-cant improvement in body mass index and lung index of rats,decreased levels of IL-18 and IL-1 β inflammatory factors(P<0.01),improved mitochondrial structure,reduced degree of fibrosis,and decreased expression of caspase-1,GSDMD,NLRP3,COL-Ⅰ,COL-Ⅲ,and α-SMA proteins in lung tissue(P<0.01).Conclusion RAS-RA has an inhibitory effect on radiation-in-duced pulmonary fibrosis,and its mechanism may be related to the inhibition of pyroptosis through the regu-lation of the NLRP3/caspase-1/GSDMD signaling pathway.
		                        		
		                        		
		                        		
		                        	
7.Feasibility of Utilizing Virtual Non-Contrast Images Derived from Spectral CT for Pulmonary Angiography with Low Contrast Medium Intake to Replace True Non-Contrast Images of Chest
Xin FANG ; Yijun LIU ; Xu WANG ; Beibei LI ; Jian JIANG ; Jingyi ZHANG
Chinese Journal of Medical Imaging 2024;32(9):956-960
		                        		
		                        			
		                        			Purpose To explore the feasibility of using virtual non-contrast images derived from spectral CT for pulmonary angiography(CTPA)to replace true non-contrast scan of chest,in order to reduce radiation dose.Materials and Methods Fifty-six patients undergoing CTPA examination in the First Affiliated Hospital of Dalian Medical University were prospectively enrolled.All patients underwent both conventional chest CT and spectral CTPA scan.On both virtual unenhanced images and true unenhanced images,CT values and standard deviation values of the pulmonary parenchyma in the level of thoracic aorta,main pulmonary artery,erector spine,as well as pulmonary apex,tracheal carina,and lung base were measured.Signal noise ratio and contrast noise ratio of thoracic aorta and pulmonary artery were calculated.The image quality and iodine residue of the two groups was evaluated.The consistency of subjective scores was also assessed.Dose length product,CT dose index volume and contrast agent dosage were recorded.The CT value,signal noise ratio,contrast noise ratio,radiation dose and subjective score of each region of interest in true unenhanced and virtual unenhanced images were compared.Results The inter-observer subjective scores were consistent(Kappa=0.984-0.992,P<0.05).The subjective scores of the chest true unenhanced and virtual unenhanced images were 4.75±0.65 and 4.65±0.49,respectively,with no statistical difference(t=0.630,P=0.437).There was no significant difference in CT values of thoracic aorta,pulmonary artery and lung tissues between true unenhanced and virtual unenhanced images(P>0.05).There was no significant difference in signal noise ratio and contrast noise ratio between pulmonary artery and thoracic aorta(P>0.05).Replacing conventional chest scan with virtual unenhanced images of CTPA reduced the radiation dose by approximately 55%,reducing from(7.27±1.74)mSv to(3.26±0.84)mSv.Conclusion It is feasible to use CTPA virtual non-contrast images to replace conventional unenhanced CT chest imaging for lung disease screening,since it can significantly reduce the radiation dose to patients.
		                        		
		                        		
		                        		
		                        	
8.Novel Immune-related Proteins Identified from Mytilus coruscus by Hemocytes Full-length Transcriptome and Serum Differential Proteome
Wen-Hui XIAO ; Hao-Dong WANG ; Zong-Xin YANG ; Fang SONG ; Yue WANG ; Jian-Yu HE ; Xiao-Lin ZHANG ; Xiao-Jun YAN ; Zhi LIAO
Chinese Journal of Biochemistry and Molecular Biology 2024;40(7):947-963
		                        		
		                        			
		                        			Mytilus is one of bivalves with great economic and ecological values.The innate immune de-fense of Mytilus shows great significance in the study of marine biological immunology.Hemolymph is the main immune tissue for Mytilus.The Nanopore full-length transcriptome of Mytilus coruscus hemocytes,and the serum differential proteomics based on SDS-PAGE analysis were performed to identify key pro-teins involving in the immune response of Mytilus hemolymph in response of different bacteria and fungi stresses.A total of 44 proteins were identified in the serum induced by different microorganisms.Among them,26 proteins showed significant differential expression level in response to different microbial stres-ses,and their functions were involved in protein folding protection,cell autophagy and apoptosis regula-tion,reactive oxygen species production,energy metabolism regulation,cell detoxification,and immune regulation.The changes in expression levels of these proteins varied in response to different bacterial and fungal stresses,suggesting that Mytilus has different immune response strategies to different bacterial and fungal stresses.The results provide a new scientific basis for understanding the differential immune mech-anism of Mytilus innate immune system in response to different types of microbial invasion,as well as the screening of specific biomarker proteins for microbial infection,and provide ideas for the healthy develop-ment and disease prevention of shellfish aquaculture.
		                        		
		                        		
		                        		
		                        	
9.Correlation of miR-155 Expression with Drug Sensitivity of FLT3-ITD+Acute Myeloid Leukemia Cell Line and Its Mechanism
Ling-Yan WANG ; Pei-Fang JIANG ; Jia-Zheng LI ; Yan-Xin CHEN ; Jian-Da HU
Journal of Experimental Hematology 2024;32(2):395-401
		                        		
		                        			
		                        			Objective:To investigate the correlation of miR-155 expression with drug sensitivity of FLT3-ITD+acute myeloid leukemia(AML)cell line and its potential regulatory mechanism.Methods:By knocking out miR-155 gene in FLT3-ITD+AML cell line MV411 through CRISPR/Cas9 gene-editing technology,monoclonal cells were screened.The genotype of these monoclonal cells was validated by PCR and Sanger sequencing.The expression of mature miRNA was measured by RT-qPCR.The treatment response of doxorubicin,quizartinib and midostaurin were measured by MTT assay and IC50 of these drugs were calculated to identify the sensitivity.Transcriptome sequencing was used to analyze change of mRNA level in MV411 cells after miR-155 knockout,gene set enrichment analysis to analyze change of signaling pathway,and Western blot to verify expressions of key molecules in signaling pathway.Results:Four heterozygotes with gene knockout and one heterozygote with gene insertion were obtained through PCR screening and Sanger sequencing.RT-qPCR results showed that the expression of mature miR-155 in the monoclonal cells was significantly lower than wild-type clones.MTT results showed that the sensitivity of MV411 cells to various anti FLT3-ITD+AML drugs increased significantly after miR-155 knockout compared with wild-type clones.RNA sequencing showed that the mTOR signaling pathway and Wnt signaling pathway were inhibited after miR-155 knockout.Western blot showed that the expressions of key molecules p-mTOR,Wnt5α and β-catenin in signaling pathway were down-regulated.Conclusion:Drug sensitivity of MV411 cells to doxorubicin,quizartinib and midostaurin can be enhanced significantly after miR-155 knockout,which is related to the inhibition of multiple signaling pathways including mTOR and Wnt signaling pathways.
		                        		
		                        		
		                        		
		                        	
10.Protective Effect of Endogenous ω-3 Polyunsaturated Fatty Acid Against Cisplatin-Induced Myelosuppression
Qi-Hua XU ; Zong-Meng ZHANG ; Chao-Feng XING ; Han-Si CHEN ; Ke-Xin ZHENG ; Yun-Ping MU ; Zi-Jian ZHAO ; Fang-Hong LI
Journal of Experimental Hematology 2024;32(5):1601-1607
		                        		
		                        			
		                        			Objective:To investigate the protective effect of endogenous ω-3 polyunsaturated fatty acid(PUFA)against cisplatin-induced myelosuppression and the mechanism of reducing apoptosis in bone marrow nucleated cells using mfat-1 transgenic mice.Methods:The experimental animals were divided into 4 groups:wild-type mice normal control group,mfat-1 transgenic mice normal control group,wild-type mice model group and mfat-1 transgenic mice model group.The mice in the model group were injected intraperitoneally with 7.5 mg/kg cisplatin on day 0 and day 7 to construct a myelosuppression model,while the mice in the normal control group were injected intraperitoneally with an equal amount of saline,and their status was observed and their body weight was measured daily.Peripheral blood was taken after 14 day for routine blood analysis,and the content and proportion of PUFA in peripheral blood were detected using gas chromatography.Bone marrow nucleated cells in the femur of mice were counted.The histopathological changes in bone marrow were observed by histopathological staining.The apoptosis of nucleated cells and the expression level changes of apoptosis-related genes in the bone marrow of mice were detected by flow cytometry and fluorescence quantitative PCR.Results:Compared with wild-type mice,mfat-1 transgenic mice showed significantly increased levels of ω-3 PUFA in peripheral blood and greater tolerance to cisplatin.Peripheral blood analysis showed that endogenous ω-3 PUFA promoted the recovery of leukocytes,erythrocytes,platelets and haemoglobin in peripheral blood of myelosuppressed mice.The results of HE staining showed that endogenous ω-3 PUFA significantly improved the structural damage of bone marrow tissue induced by cisplatin.Flow cytometry and PCR showed that,compared with wild-type mice model group,the apoptosis rate of bone marrow nucleated cells in mfat-1 transgenic mice was significantly reduced(P<0.001),and the expression of anti-apoptotic genes Bcl-2 mRNA was significantly increased(P<0.01),while the expressions of pro-apoptotic genes Bax and Bak mRNA were significantly reduced(P<0.001,P<0.05).Conclusion:Endogenous ω-3 PUFA can reduce cisplatin-induced apoptosis in bone marrow nucleated cells,increase the number of peripheral blood cells and exert a protective effect against cisplatin-induced myelosuppression by regulating the expression of apoptosis-related genes.
		                        		
		                        		
		                        		
		                        	
            
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