1.Effects of D-ribose on High-energy Phosphate Metabolism of Skeletal Muscle Tissues of Tired Mice
Yan DING ; Dan WU ; Zhanhong JIA ; Dandan LI ; Yun WEI ; Jinxin RUAN ; Shuofeng ZHANG ; Yikun SUN
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(9):1916-1920
This article was aimed to study effect of D-ribose on the high-energy phosphate metabolism of skeletal muscle tissues of tired mice. The model was made by burden swimming. And then, the mice were divided into four groups, which were the model group, D-ribose group, caffeine group, and D-ribose with caffeine group). Intragastric administrations of drugs were given to all mice in four groups, three times per day. And all mice continued to swim for three days. The time of swimming was recorded. Gastrocnemius of mice were removed after swimming or 3 days later to measure the concentration of ATP, ADP, AMP and IMP with the HPLC. The results showed that compared with the control group, the time of burden s wimming was significantly prolonged for mice in the D-ribose group and the D-ribose with caffeine group. After three-day recovery, the concentration of ATP, AMP and IMP of gastrocnemius in the D-ribose group and the D-ribose with caffeine group mice was significantly increased. There was no significant difference in the caffeine group mice. It was concluded that D-ribose is involved in the high-energy phosphate metabolism of skeletal muscle tissues of tired mice . D-ribose promotes the recovery of ATP concentration in the gastrocnemius of tired mice, and prolongs the time of burden swimming. Therefore, it has a certain anti-fatigue effect .
2.Changes of DDAs content affected by different processing time and its relationship with safety of processed Fuzi.
Zhiyong LI ; Shuofeng ZHANG ; Hongsheng CHANG ; Jianning SUN ; Fei LI
China Journal of Chinese Materia Medica 2009;34(9):1086-1089
OBJECTIVETo study the correlation between content changes of Diester diterpenoid alkaloids (DDAs) content and safety of the processed Fuzi.
METHODSequential and Bliss methods were used to evaluate the safety of 7 kinds of Fuzi processed with different processing time. The relationship between ED50, TD50, TI and content changes of DDAs of those processed Fuzi was studied, the correlation between the content changes and effect of different processed Fuzi was analyzed, and the toxicity of those processed Fuzi with multiple linear regression was tested.
RESULTFuzi with good efficiency and safety contains proper hypaconitine (HA) and mesaconitine (MA). Aconitine (AC) interfered efficacy of Fuzi (negative correlation), HA showed positive correlation with toxicity and efficacy of Fuzi.
CONCLUSIONHA and MA kept in determinate proportion are very important for the safety and effectivity of processed Fuzi.
Aconitine ; analogs & derivatives ; metabolism ; Alkaloids ; chemistry ; Animals ; Diterpenes ; chemistry ; metabolism ; Drug-Related Side Effects and Adverse Reactions ; Drugs, Chinese Herbal ; adverse effects ; metabolism ; pharmacology ; Magnoliopsida ; chemistry ; Male ; Rats ; Time Factors
3.Study on intestinal absorption of multiple constituents under the drug response of Wuwei-Jiangya recipe
Huihui ZHAO ; Li YU ; Kailun ZHANG ; Changling WEI ; Yang LIU ; Baosheng ZHAO ; Shuofeng ZHANG ; Xiaoyan GAO ; Liying LIU
International Journal of Traditional Chinese Medicine 2012;34(9):804-807
ObjectiveTo research the intestinal absorption characteristics in rats of multiple constituents,when Wuwei-Jiangya recipe was used in rats and showed reducing blood pressure effects.Methods ① After extracting Wuwei-Jiangya recipe,we fed it to rats once daily,until the blood pressure reduced; ②Establish Wuwei-Jiangya recipe and intestinal absorption of multiple constituents fingerprint by using reverted gut sac method and RP-HPLC fmgerprint.ResultsAfter one week's administration,there was the hypotensive effects and multiple constituents can be absorbed by intestine.ConclusionWhen the drug works,reverted gut sac method for studying intestinal absorption constituents can be used for locking the exposure constituents.
4.Preparation of polyvinyl alcohol/lota-carrageenan scaffolds and its biocompatibility
Jing CUI ; Yabin ZHANG ; Siqi MA ; Yanjie XIONG ; Man CUI ; Shuofeng LI ; Pengcheng CHE ; Fanglian YAO ; Hong SUN
Chinese Journal of Tissue Engineering Research 2017;21(2):215-220
BACKGROUND:Polyvinyl alcohol (PVA) hydrogel with similar porous structure and mechanical properties to the natural cartilage is very suitable for the repair of articular cartilage. However, the pure PVA hydrogel after lyophilization wil be accompanied by the shrinkage of the polymer network and the col apse of the pores, leading to the inhomogeneous performance of the material even in the state of re-swel ing. Addition of the active polymer wil increase the cel adhesion ability of PVA hydrogel. OBJECTIVE:To construct PVA/lota-carrageenan (l-CA) composite materials with different mass fractions of l-CA and evaluate the biocompatibility with vascular endothelial cel s. METHODS:PVA/l-CA composite films with different contents of l-CA were fabricated and then co-cultured with vascular endothelial cel s. Attachment, proliferation and morphological changes of vascular endothelial cel s on the composite were observed by scanning electron microscope and MTT assay to evaluate its biocompatibility. PVA/l-CA three-dimensional scaffold with different contents of l-CA were constructed, and hemolysis experiment was conducted according to the biological evaluation standards of medical devices, and the porosity and pore size were observed using scanning electron microscope. RESULTS AND CONCLUSION:In vitro experimental results showed that the addition of l-CA could significantly increase the biological activity of PVA hydrogel, and promote the cel attachment and proliferation on the scaffold. The hemolysis rate of each experimental group was less than 5%(the accepted safety standard), suggesting that the composite materials were in accordance with the standard of medical devices for hemolysis experiment. These findings indicate that the composite scaffolds with 20%-30%l-CA possess the pore size suitable for cel growth and proliferation and the porosity beneficial for transportation of nutrients and metabolites, which can serve as an excel ent scaffold for tissue engineering.
5.Effects of biomimetic network membrane prepared by chitosan/gelatin/pectin on proliferation and mineralization of mesenehymal stem cells
Hong SUN ; Zhiwen YAN ; Shuofeng LI ; Yanjie XIONG ; Fan LIANG ; Ao LI ; Fanglian YAO ; Pengcheng CHE
Journal of Jilin University(Medicine Edition) 2019;45(1):17-22,后插1
Objective::To explore the effects of the biomimetic network membrane prepared by chitosan/gelatin/pectin on the proliferation and mineralization of mesenchymal stem cells (MSCs) , and to evaluate its feasibility of constructing tissue engineering bone.Methods:Chitosan, gelatin and pectin were made into a new biomimetic network membrane in a certain ratio by biomimetics.The experiment was divided into control group (MSCs+conventional medium) , material group (MSCs+network membrane+conventional medium) and material+OS group (MSCs+network membrane+OS medium) .The cell morphology was observed by inverted phase contrast microscope;the growth and secretion of extracellular matrix of the MSCs were observed under scanning electron microscope (SEM) .The proliferation of cells was determined by MTT assay (The MSCs were divided into negative control group and material group, and they were cultivated with blank medium and medium including materials) .The expression of calcium in MSCs was detected by Alizarin Red staining.Real-time polymerase chain reaction (RT-PCR) was used to determine the expression levels of osteocalcin (OC) mRNA and osteopontin (OPN) mRNA in the MSCs.Results:The network membrane was semitransparent thin film.The MSCs were short shuttle and clustered under inverted phase contrast microscope.After cultured for 7d, the MSCs were shuttle;after cultured for 14d, the number of MSCs was increased, with pseudo feet on the membrane;after cultured for21d, the MSCs clustered with a lot of neo-formed extracellular matrix.The MTT results showed that there was no significant difference in the proliferation level of MSCs between material group and negative control group (P>0.05) .The Alizarin Red staining results showed that the MSCs in the network membrane were dyed orange red.The RT-PCR results showed that the expression levels of OC mRNA in the MSCs in material group and material+OS group were lower on the 7th and 14th days, but on the 21th day, the expression levels were significantly increased and reached the peak;the expression level of OC mRNA in the MSCs in material group was significantly increased on the 7th day, and the expression level reached the peak on the 14th day, then fell slightly on the 21th day;compared with control group, the expression levels of OC mRNA and OPN mRNA in the cells in material group and material+OS group at different time points were significantly increased (P<0.01) , but there were no significant differences between material group and material+OS group (P>0.05) .Conclusion:Chitosan/gelatin/pectin biomimetic network membrane has good biocompatibility, and MSCs can grow and proliferate well on the membrane.The membrane can induce the MSCs to express mineralization-related genes and proteins without inducers.
6.Clinical evaluation of a melting curve analysis-based PCR assay for glucose phosphate dehydrogenase gene mutation detection.
Tizhen YAN ; Qingyan ZHONG ; Ning TANG ; Shuofeng WEI ; Qiuying HUANG ; Shiqiang LUO ; Wugao LI ; Qiuhua WANG ; Ren CAI
Chinese Journal of Medical Genetics 2014;31(2):156-162
OBJECTIVETo evaluate the clinical value of multicolor melting curve analysis(MMCA) for detecting genetic mutations in G6PD deficiency.
METHODSA total of 402 peripheral blood samples(256 males and 146 females) were collected from suspected patients or their relatives at the Prenatal Diagnosis Center of Liuzhou Maternal and Child Health Hospital between March 2012 and May 2012. The samples were screened by G6PD/6PGD quantitative ratio testing. The reliability of the assay was evaluated by multiplex probe melting curve assay(which can detect 16 G6PD mutations) and DNA sequencing through a double blind study.
RESULTSOne hundred seventy cases with G6PD/6PGD ratio < 1.0 and 232 cases with G6PD/6PGD ratio ≥ 1.0 were detected by the enzymological method. DNA sequencing has identified 182 wild type samples, 151 hemizygous mutation samples, 5 female homozygous mutation samples, 54 female heterozygous mutation samples and 10 female double heterozygous mutation samples. Multicolor melting curve analysis has detected 185 wild type samples, 148 hemizygous mutation samples, 5 female homozygous mutation samples, 55 female heterozygous mutation samples and 9 female double heterozygous mutation samples. The specificity and sensitivity of G6PD gene mutation detection by multicolor melting curve analysis were 100%(182/182) and 98.6%(217/220), respectively. The positive predictive value and negative predictive value were 99.5%(216/217) and 98.4%(182/185), respectively, and the Youden's index was 0.986. The concordance rate of the sample detection between the melting curve assay and DNA sequencing was 99.0%(398/402). Twenty-one different genotypes were detected by the multicolor melting curve analysis and 24 different genotypes were detected by DNA sequencing. Four samples containing mutations(c.196T>A or c.406C>T) were not detected by multicolor melting curve analysis, which can be attributed to different technical settings of the two methods.
CONCLUSIONMulticolor melting curve analysis for G6PD gene mutation detection is a simple, rapid, sensitive and specific method, which can be used for clinical diagnosis of G6PD deficiency.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Humans ; Infant ; Infant, Newborn ; Male ; Mutation ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA
7.Effect of cartilage tissue engineering scaffolds PVA/ι-CA on biological behavior and biocompatibility of ATDC-5 cells
Pengcheng CHE ; Xuan CHE ; Shuofeng LI ; Yabin ZHANG ; Yanjie XIONG ; Man CUI ; Jing CUI ; Fanglian YAO ; Hong SUN
Journal of Jilin University(Medicine Edition) 2017;43(6):1092-1097,前插2-前插3
Objective: To investigate the effect of cartilage tissue engineering scaffold PVA/ι-CA on the biological behavior of the ATDC-5 cells,and to evaluate its feasibility on constructing tissue engineering cartilage. Methods:The polyvinyl alcohol (PVA)and carrageenan were used to make the composite scaffold material PVA/ι-CA according to a certain proportion by physical blending technology and repeated freezing thawing method,and the porosity and pore size of PVA/ι-CA were detected.The ATDC-5 cells were seeded into the composite scaffold and its growth was observed; the expressions of collagen type Ⅱ in the ATDC-5 cells were tested by immunohistochemical staining and immunofluorescence staining; the morphology of the ATDC-5 cells was confirmed by Toluidine blue staining.The growth and secretion of extracellular matrix of the ATDC-5 cells were observed under scanning electron microscope (SEM);the proliferative rates of ATDC-5 cells in composite scaffold materials in negative control group (added with DMEM culture media)and experimental group (added with DMEM contain scaffold)were determined by MTT assay.The composite scaffolds were implanted subcutaneously in the SD rats.The histocompatibility and vascularization in vivo of the composite scaffolds were evaluated.Results:The average porosity of cartilage tissue engineering scaffold PVA/ι-CA was (86.88±3.88)%,and the average pore size was 20-40 μm.The HE staining results showed that the ATDC-5 cells grew well with the polygon and plumpness morphology. All the samples were stained positive for collagen type Ⅱ by immunohistochemistry and immunofluorescence staining,which verified the normal phenotype of chondrocytes on the scaffolds. All the sample were stained positive for toluidine blue staining,which verified ECM deposition of the ATDC-5 cells on the scaffolds.The number of the positive cells was significantly increased with the prolongation of time.After cultured for 7 d,few of the ATDC-5 cells presented polygonal;after cultured for 14 d,the ATDC-5 cells distributed more densely,and contacted with each other on the scaffold;after cultured for 21 - 28 d,the ATDC-5 cells filled the interconnected pores of the scaffolds,synthesizing a significant amount of neo-formed ECM.The proliferation of ATDC-5 cells in PVA/ι-CA grew fast during 7-14 d,and it became slow during 21-28 d;the difference was not statistically significant compared with control group (P >0.05).The subcutaneous implantation results showed the inflammatory reactions were slight at the early stage and eviated gradually,there was an increasing angiogenesis at the late stage,and the degradation and absorption of the meterial were slight.Conclusion:PVA/ι-CA composite material will be an ideal material for the cartilage tissue engineering.
8. Comparison of curative effect of different surgical methods on varicocele
Hongzhi LIU ; Hua PENG ; Shuofeng LI ; Yongshuang XIAO ; Yue CHEN ; Hailong LI ; Rumin WEN
International Journal of Surgery 2020;47(1):35-40
Objective:
To compare the curative effect of three different surgical methods: peritoneal varicocele ligation, peritoneal single-port laparoscopy and microscopy on varicocele.
Methods:
Retrospective analysis of the case data of 150 patients with varicocele treated in the Affiliated Hospital of Xuzhou Medical University from September 2016 to September 2018. The average age was 24.5 years, and the age range was 22-30 years. The patients were divided into three groups according to different surgical methods: open group, laparoscopy group and microscope group, with 50 cases in each group. Patients in the open group were treated with retroperitoneal spermatic cord ligation. Patients in the laparoscopic group were treated with single-hole laparoscopic laparoscopic surgery. Patients in the microscope group were treated with microscope surgery. Operation time, postoperative hospitalization time, hospitalization cost reserved arteries, surgical complications (such as testicular hydrocele, scrotal edema, epididymitis, testicular atrophy), recurrence, and semen quality improvement were compared between three groups. Measurement data were expressed as mean ± standard deviation(
9.A hnRNPA2B1 agonist effectively inhibits HBV and SARS-CoV-2 omicron in vivo.
Daming ZUO ; Yu CHEN ; Jian-Piao CAI ; Hao-Yang YUAN ; Jun-Qi WU ; Yue YIN ; Jing-Wen XIE ; Jing-Min LIN ; Jia LUO ; Yang FENG ; Long-Jiao GE ; Jia ZHOU ; Ronald J QUINN ; San-Jun ZHAO ; Xing TONG ; Dong-Yan JIN ; Shuofeng YUAN ; Shao-Xing DAI ; Min XU
Protein & Cell 2023;14(1):37-50
The twenty-first century has already recorded more than ten major epidemics or pandemics of viral disease, including the devastating COVID-19. Novel effective antivirals with broad-spectrum coverage are urgently needed. Herein, we reported a novel broad-spectrum antiviral compound PAC5. Oral administration of PAC5 eliminated HBV cccDNA and reduced the large antigen load in distinct mouse models of HBV infection. Strikingly, oral administration of PAC5 in a hamster model of SARS-CoV-2 omicron (BA.1) infection significantly decreases viral loads and attenuates lung inflammation. Mechanistically, PAC5 binds to a pocket near Asp49 in the RNA recognition motif of hnRNPA2B1. PAC5-bound hnRNPA2B1 is extensively activated and translocated to the cytoplasm where it initiates the TBK1-IRF3 pathway, leading to the production of type I IFNs with antiviral activity. Our results indicate that PAC5 is a novel small-molecule agonist of hnRNPA2B1, which may have a role in dealing with emerging infectious diseases now and in the future.
Animals
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Mice
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Antiviral Agents/pharmacology*
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COVID-19
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Hepatitis B virus
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Interferon Type I/metabolism*
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SARS-CoV-2/drug effects*
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Heterogeneous-Nuclear Ribonucleoprotein Group A-B/antagonists & inhibitors*
10.Targeting papain-like protease for broad-spectrum coronavirus inhibition.
Shuofeng YUAN ; Xiaopan GAO ; Kaiming TANG ; Jian-Piao CAI ; Menglong HU ; Peng LUO ; Lei WEN ; Zi-Wei YE ; Cuiting LUO ; Jessica Oi-Ling TSANG ; Chris Chun-Yiu CHAN ; Yaoqiang HUANG ; Jianli CAO ; Ronghui LIANG ; Zhenzhi QIN ; Bo QIN ; Feifei YIN ; Hin CHU ; Dong-Yan JIN ; Ren SUN ; Jasper Fuk-Woo CHAN ; Sheng CUI ; Kwok-Yung YUEN
Protein & Cell 2022;13(12):940-953
The emergence of SARS-CoV-2 variants of concern and repeated outbreaks of coronavirus epidemics in the past two decades emphasize the need for next-generation pan-coronaviral therapeutics. Drugging the multi-functional papain-like protease (PLpro) domain of the viral nsp3 holds promise. However, none of the known coronavirus PLpro inhibitors has been shown to be in vivo active. Herein, we screened a structurally diverse library of 50,080 compounds for potential coronavirus PLpro inhibitors and identified a noncovalent lead inhibitor F0213 that has broad-spectrum anti-coronaviral activity, including against the Sarbecoviruses (SARS-CoV-1 and SARS-CoV-2), Merbecovirus (MERS-CoV), as well as the Alphacoronavirus (hCoV-229E and hCoV-OC43). Importantly, F0213 confers protection in both SARS-CoV-2-infected hamsters and MERS-CoV-infected human DPP4-knockin mice. F0213 possesses a dual therapeutic functionality that suppresses coronavirus replication via blocking viral polyprotein cleavage, as well as promoting antiviral immunity by antagonizing the PLpro deubiquitinase activity. Despite the significant difference of substrate recognition, mode of inhibition studies suggest that F0213 is a competitive inhibitor against SARS2-PLpro via binding with the 157K amino acid residue, whereas an allosteric inhibitor of MERS-PLpro interacting with its 271E position. Our proof-of-concept findings demonstrated that PLpro is a valid target for the development of broad-spectrum anti-coronavirus agents. The orally administered F0213 may serve as a promising lead compound for combating the ongoing COVID-19 pandemic and future coronavirus outbreaks.
Animals
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Coronavirus Papain-Like Proteases/antagonists & inhibitors*
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Cricetinae
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Humans
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Mice
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Pandemics
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SARS-CoV-2/enzymology*
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COVID-19 Drug Treatment