1.Role of Ferroptosis in Non-small Cell Lung Cancer and Progress of Traditional Chinese Medicine Intervention
GUO XIAOQI ; WANG TIANQI ; XIA JINCHAN ; ZENG HUAHUI ; SHI WENBO
Chinese Journal of Lung Cancer 2024;27(3):216-230
Non-small cell lung cancer(NSCLC)is one of the malignant tumors with high morbidity and mortality worldwide.Ferroptosis is a new type of programmed cell death caused by abnormal accumulation of iron-dependent reactive oxygen species(ROS)leading to lipid peroxidation.It involves the balance between iron metabolism,lipid metabolism,oxy-gen free radical reaction and lipid peroxidation.Recent studies have found that ferroptosis is closely related to the occurrence and development of NSCLC.Due to the emergence of chemotherapy resistance and radiotherapy resistance in the treatment of NSCLC,there is an urgent need to develop new effective drugs and treatment strategies.Traditional Chinese medicine has unique advantages in the prevention and treatment of NSCLC due to its multi-targets and minimal side effects.In this review,we summarize the mechanism of ferroptosis in NSCLC,and discuss the research status of active ingredients of traditional Chinese medicine,single-herb traditional Chinese medicine and Chinese herbal compounds in the intervention of NSCLC through ferroptosis,in order to provide a new theoretical basis for the research of ferroptosis pathway and the prevention and treatment of NSCLC by targeted ferroptosis of traditional Chinese medicine.
2.Construction of a recombinant Bacillus subtilis strain expressing SpaA and CbpB of Erysipelothrix rhusiopathiae and evaluation of the strain immunogenicity in a mouse model.
Zhonglin CHENG ; Hao HUANG ; Siyi CAO ; Huahui SHI ; Jiye GAO ; Jixiang LI
Chinese Journal of Biotechnology 2024;40(12):4521-4532
To construct a recombinant Bacillus subtilis strain expressing SpaA and CbpB of Erysipelothrix rhusiopathiae for oral administration, we constructed the recombinant plasmid pDG1730-CBJA by fusion PCR and seamless cloning. The plasmid was introduced into B. subtilis KC strain by natural transformation, and the recombinant strain KC-spaA-cbpB was screened out on the plate containing spectinomycin (sper) and confirmed by PCR and starch degradation test. The SpaA and CbpB expressed by KC-spaA-cbpB were detected by Western blotting and indirect immunofluorescence assay, and the genetic stability of the recombinant strain in mice was determined. The plasmid pMAD-∆sper with knockout of sper was constructed and transformed into KC-spaA-cbpB. The sper-deleted mutant strain KC-spaA-cbpB: : ∆sper was screened and identified, and its immunogenicity in a mouse model was evaluated by oral immunization. The results showed that the recombinant strain KC-spaA-cbpB was stable in mice, expressing SpaA on the cell surface and CbpB on the spore surface. KC-spaA-cbpB: : ∆sper expressed SpaA and CbpB. The mice vaccinated with the spores of KC-spaA-cbpB: : ∆sper had higher levels of SpaA and CbpB-specific IgG in the serum that those vaccinated with the wild-type spores 42 days after vaccination by gavage (P < 0.01). The protective rate of mice immunized with the recombinant spores was 67.5%. The results indicated that a recombinant B. subtilis strain expressing SpaA and CbpB of E. rhusiopathiae was successfully constructed, and the recombinant strain laid a foundation for the development of oral live vector vaccines for swine erysipelas.
Animals
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Bacillus subtilis/immunology*
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Mice
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Erysipelothrix/immunology*
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Bacterial Proteins/immunology*
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Bacterial Vaccines/genetics*
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Erysipelothrix Infections/prevention & control*
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Immunization
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Mice, Inbred BALB C
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Plasmids/genetics*
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Immunogenicity, Vaccine
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Administration, Oral
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Antigens, Bacterial
3.Study on preparation and in vivo safety of progesterone cocrystal
Wenbo SHI ; Man LI ; Huahui ZENG ; Hui ZHANG ; Wenwen ZHAO ; Xiangxiang WU
China Pharmacy 2023;34(13):1567-1572
OBJECTIVE To prepare progesterone-2-chloro-4-nitroaniline cocrystal (CNA) so as to improve the solubility of progesterone and primarily evaluate the safety of the progesterone cocrystal in vivo. METHODS Using progesterone as the main body and CNA as the ligand, progesterone-CNA cocrystal was prepared with solvent evaporation method. The cocrystal was characterized by X-ray single crystal diffraction, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (IR). The dissolution rate of cocrystal was compared with those of progesterone and physical mixture. Forty-eight female KM mice were randomly divided into normal group (phosphate buffer containing 0.1% dimethyl sulfoxide), progesterone group (16 mg/kg), CNA group (9 mg/kg), progesterone-CNA cocrystal low-dose, medium- dose and high-dose groups (6, 12.5, 25 mg/kg), with 8 mice in each group. They were given relevant medicine/solvent intramuscularly, once a day, for consecutive 14 d. The safety of cocrystal was evaluated primarily by determining/observing the changes in body weight, organ index, tissue morphology, blood routine indicators, and liver and kidney function indicators. RESULTS The new crystal structure in the X-ray single crystal diffraction results, the new characteristic peak in the XRPD pattern, the change of melting point in the DSC results, and the change of the characteristic peak position in the range of 3 500- 2 750 cm-1 and 1 700-1 250 cm-1 in the infrared spectrum all Δ 基金项目国家重点研发计划项目(No.2022YFC3502100) indicated that progesterone-CNA cocrystal was successfully *第一作者 硕士研究生 。研究方向 :药物制备技术与工艺 。 prepared, and the dissolution rate of cocrystal was more than E-mail:SWB_1221@163.com # 通信作者教授,硕士生导师,博士。研究方向:药物制备技术与 twice that of the progesterone raw material drug. The results of 工艺。E-mail:wuxx-415@126.com in vivo safety experiments showed that the mortality rate of all 中国药房 2023年第34卷第13期 China Pharmacy 2023 Vol. 34 No. 13 · 1567 · groups was zero. Compared with normal group, uterine indexes of mice in progesterone group and progesterone-CNA cocrystal groups were significantly increased (P>0.05), and endometrium was also thickened; there was no statistical difference in the changes of body mass, liver and kidney function, liver index, kidney index, the number of leukocyte, lymphocyte and neutrophil in routine blood test among those groups (P>0.05), and the morphology of liver and kidney tissue has also no significant difference. However, the number of plasma red blood cells in the progesterone group decreased significantly (P<0.05), and there was no statistical significance in the number difference of red blood cells among progesterone-CNA cocrystal groups (P>0.05). CONCLUSIONS The progesterone-CNA cocrystal is successfully prepared with good safety in vivo, which significantly improve the solubility of progesterone.
4.Study on Protective Effects of Crocin against Triptolide-induced Visceral Organ Injury in Mice
Yinyin YAN ; Min YAN ; Xiangxiang WU ; Xin ZHU ; Wenbo SHI ; Mengyuan JIANG ; Huahui ZENG
China Pharmacy 2021;32(19):2320-2326
OBJECTIVE:To study the prot ective effects of crocin (CR)against triptolide (TP)-induced visceral organ injury in mice,and to provide reference for the studying TP compatibility and detoxification. METHODS :Fifty mice were randomly divided into normal group ,TP low-dose and high-dose groups (i.e. TP-L group ,TP-H group ,with 300,600 μg/kg),TP low-dose and high dose combined with CR groups (i.e. TP-L+CR group ,TP-H+CR group ,with 300 μg/kg TP+100 mg/kg CR ,600 μg/kg TP+ 100 mg/kg CR ),with 10 mice in each group. Except for normal group ,other groups were given relevant medicine intragastrically , once a day ,for consecutive 7 d. The body weight of mice was weighted every day ,and their death was recorded. After last administration,the mice were sacrificed ,and the heart ,liver,kidney and testis were taken ,and the organ index was calculated ; serum levels of ALT ,AST,BUN and Scr ,the activity of T-SOD and the contents of MDA were all determined. The pathological changes of heart ,liver,kidney and testis were observed ;mRNA expression of Bcl- 2,Bax and caspase- 3 in liver tissue were determined. RESULTS :Three,five,two and three mice in TP-L group ,TP-H group ,TP-L+CR group and TP-H+CR group died respectively,and the survival rates were 70%,50%,80% and 70%,respectively. Compared with normal group ,the body weight (7th day of experiment ),heart index ,liver index ,kidney index (except for TP-L group ),testicular index ,T-SOD activity and mRNA expression of Bcl- 2 in liver tissue ,serum levels of ALT (except for TP-L group ),AST(except for TP-L group ),BUN and Scr,MDA content and mRNA expression of Bax ,mRNA expression of caspase- 3 in liver tissue were increased significantly (P< 0.05 or P<0.01). There were obvious pathological changes in heart ,liver,kidney and testis tissue. Compared with the same dose of TP alone group ,the above indexes of TP combined with CR group were improved in varying degrees. Except for the renal index and serum ALT level of TP-L+CR group ,there was statistical significance for all indexes (P<0.05 or P<0.01);the pathological injuries of heart ,liver,kidney and testis were significantly improved. CONCLUSIONS :CR can relieve the damage of heart , liver,kidney and testis induced by TP ,which may be related to the antioxidant stress of CR.
5.Value of procalcitonin and lipopolysaccharide in identifying pathogens and evalu-ating therapeutic efficacy of hospital-acquired pneumonia
Yongxin SHI ; Weiqing SONG ; Huahui LI
Chinese Journal of Infection Control 2016;(1):41-44
Objective To explore the value of procalcitonin (PCT)and lipopolysaccharide (LPS)in identifying pathogens and evaluating therapeutic efficacy of hospital-acquired pneumonia (HAP).Methods A total of 110 HAP patients were enrolled in a prospective study,patients were divided into gram-negative bacterial infected HAP group (G- infected group,n=50),gram-positive bacterial infected HAP group (G+ infected group,n=30),and control group (nontypical pathogen or virus infected group,n =30).Serum levels of PCT,LPS and C-reactive protein (CRP)of patients were dynamically detected,receiver operating characteristic (ROC)curve and area under the curve (AUC)were adopted to assess the value of PCT and LPS in predicting pathogenic bacteria causing HAP. Results PCT and LPS levels of G - infected group were (3.43 ±1 .15)ng/mL and (0.20 ±0.08)EU/mL respec-tively,which were higher than G+ infected group ([0.42±0.12]ng/mL and [0.05±0.02]EU/mL respectively)and control group([0.14±0.08]ng/mL and [0.02 ±0.01 ]EU/mL respectively)(all P <0.05 ).Levels of PCT and CRP of G- infected group before and after therapy were both significantly different ([3.43±1 .15]ng/mL vs [0.63 ±0.22]ng/mL,[47.26±30.35]mg/L vs [9.21 ±6.54]mg/L,respectively)(both P <0.01).The levels of PCT, LPS,and CRP in moderate and severe patients were all significantly higher than mild patients ([5.43±1 .05]ng/mL vs [0.72±0.32]ng/mL,[0.33±0.07]EU/mL vs [0.09 ±0.04]EU/mL,[57.46 ±20.15 ]mg/L vs [8.25 ± 5.24]mg/L,respectively)(all P <0.05).Sensitivity and specificity of combined detection of PCT and LPS in dif-ferentiating gram-negative bacteria infected VAP from gram-positive bacteria infected VAP were 95.83% and 96.15% respectively,AUC was 0.95.Conclusion PCT and LPS have certain value in identifying pathogens of HAP,combined detection of PCT and LPS can increase specificity in identifying HAP type,and assess the efficacy of antimicrobial therapy in accordance with the dynamic change.

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