1.A Research Progress on Matrine and Its Preparations
Huan HE ; Xinyue SHANG ; Shaojiang SONG
World Science and Technology-Modernization of Traditional Chinese Medicine 2016;18(7):1099-1107
Matrine is an effective natural product isolated from the traditional herb,Sophora flavescens,with wide range of pharmacological and clinical effects and treating as the treatment for chronic viral hepatitis in clinic.Currently,the mainstream clinical preparations of matrine involved injections,tablets,capsules and suppositories,while preparations under development included targeting preparations,sustained-release preparations and transdermal drug delivery preparations.Admittedly,It can significantly improve the biological utilization of matrine through the optimization of its clinical preparations for with scientific researches and the development of new dosage forms.While the clinical applications will be expanded and the clinical efficacy of the preparations of matrine enhanced with the development of researches for new dosage forms and drugs and the application of new materials.It may be also benefical to the study of monomer compositions of Chinese material medica,including the development of the preparation of matrine.Rested on the recent studies,pharmacological activities,preparation methods and researches for the preparations of matrine was summarized in this text for providing a reference for developing its preparations in the future.
2.Construction and assessment of recombinant plasmid pRNAT-U6.1/CFB siRNA and its inhibitory effect on proliferation of human umbilical vein endothelial cells
Huan, TONG ; Qingli, SHANG ; Jingxue, MA ; Jian, GAO ; Xin, WANG
Chinese Journal of Experimental Ophthalmology 2015;33(8):686-690
Background Choroidal neovascularization (CNV) is one of the causes of blindness in multiple eye diseases.Researches showed that complement system participates in the pathogenesis of CNV.Objective This study was to construct the recombinant of complement factor B-small interference RNA (CFB-siRNA) expression vector and to observe its inhibitory effect on human umbilical vein endothelial cells (ECV-304).Methods CFB gene primers were designed based on human CFB gene,and an expression vector of CFB-siRNA was constructed by inserting CFB-siRNA into pRNAT-U6.1/Neo plasmid.Recombinant plasmids were confirmed by the digestion analysis of restriction endonuclease,and all inserted sequences were verified by DNA sequencing.The recombinant pRNAT-U6.1/CFB-siRNA plasmid and the blank plasmid were transfected into ECV-304 cells in the CFB-siRNA group and blank plasmid group by electroblot,respectively,and non-transfected cells served as the normal control group.The cells were observed under the fluorescence microscope 48 hours after transfection,and the transfective efficiency was calculated.The relative expression of CFB mRNA in the cells of different groups was detected by semi-quantitative reverse transcription PCR (RT-PCR).MTT was employed to calculated the growth inhibitory rates of the cells 24,48 and 72 hours after transfection.The percentages of the cells in different cell cycles were detected by flow cytometry.Results The sequence of the target vector was identical to the designed sequence.The green fluorescence protein (GFP) was seen in both the CFB-siRNA group and the blank plasmid group.The relative expression levels of CFB mRNA were 0.07 ±0.04,0.14 ±0.02 and 0.14 ±0.03 in the CFB-siRNA group,the blank plasmid group and the normal control group,respectively,a significant difference was obtained among the three groups (F=233.05,P =0.00);the expression level of CFB mRNA in the CFB-siRNA group was significantly declined in comparison with the blank plasmid group and the normal control group (both at P<0.05).The growth inhibitory rates of the cells were (23.45 ±0.01) %,(33.48 ±0.02) % and (45.49±0.01) % at 24,48 and 72 hours after transfection,respectively,a significant difference was obtained among the three groups (Fgroup =212.99,P =0.00);the growth inhibitory rates in CFB-siRNA group were significantly higher than that in the blank plasmid group and normal control group (all at P< 0.05).The percentages of G1 phase cells were (44.4 ±0.5) %,(25.8 ±0.4) % and (27.9 ± 0.6) % in the CFB-siRNA group,the blank plasmid group and the normal control group respectively,a significant difference was obtained among the three groups (F=58.98,P=0.00).The percentages of G1 phase and G2 phase cells in the CFB-siRNA group were significantly higher than those in the blank plasmid group and the normal control group (all at P<0.05).Conclusions Recombinant pRNAT-U6.1/CFB siRNA inhibits the proliferation of ECV-304 cells effectively by arresting the cells in G1 intermediate phase of the growth cycle.
3.Long-term results of dual anti-collagen membrane in guided tissue regeneration.
Shu-huan SHANG ; Cheng-zhang LI ; Ming-wen FAN
Chinese Journal of Stomatology 2010;45(6):346-349
OBJECTIVETo evaluate the long-term clinical effect of dual anti-collagen membranes in guided tissue regeneration (GTR).
METHODSThis randomized clinical trial included 26 teeth in 24 patients, presenting a total of 31 lesions consisting of intrabony defects and furcation defects. Twenty-six teeth were divided into two groups and treated by GTR with dual anti-collagen membranes and atelocollagen membranes, respectively. At baseline, 6 months, 1, 3 and 6 years, the following parameters were recorded: clinical attachment level, probing depth, gingival recession and the quantity of alveolar bone analyzed by computer assisted densitometry image analysis (CADIA).
RESULTSAt 1 year after GTR surgery, the gain of clinical attachment in dual anti-collagen membranes group was (3.93 ± 1.74) mm, compared with (2.25 ± 1.90) mm in atelocollagen group (P = 0.044). The increasing of the value of CADIA in dual anti-collagen membrane and atelocollagen group were (53.14 ± 21.35) and (32.96 ± 17.97), P = 0.031. At 3 and 6 years, clinical parameters remained basically stable in both groups, compared to that at 1 year after surgery.
CONCLUSIONSThe regeneration of periodontal tissues obtained by GTR with dual anti-collagen membranes could be maintained on a long-term basis.
Adult ; Alveolar Bone Loss ; surgery ; Bone Regeneration ; Collagen ; Densitometry ; methods ; Dental Plaque Index ; Female ; Follow-Up Studies ; Furcation Defects ; surgery ; Guided Tissue Regeneration, Periodontal ; methods ; Humans ; Image Interpretation, Computer-Assisted ; methods ; Male ; Membranes, Artificial ; Middle Aged ; Periodontal Attachment Loss ; surgery ; Periodontal Index ; Young Adult
4.Research progress of the related cytokines of Th1 and Th2 on glaucoma
Chinese Journal of Experimental Ophthalmology 2017;35(11):1052-1056
Glaucoma is a leading cause of irreversible blindness,which includes visual field defects and chronic degenerative diseases of the optic nerve,such as apoptosis of the retinal ganglion cells (RGCs) and progressive loss of optic nerve axons.Elevated intraocular pressure is the primary risk factor for glaucoma,and lowering intraocular pressure is the first choice of treatment for slowing the development of glaucoma.It has recently been found that immunological factors are one of the non-stress-dependent risk factors for optic nerve damage in glaucoma,while most of the immune and non-immune biological effects are regulated by cytokines.CD4+ helper T cells are the major sources of cytokines.Thl and Th2-related cytokines play an essential role in the pathogenesis of glaucoma and related with the survival and apoptosis of RGCs.In this review,we discussed the potential relationship between Th1/Th2-related cytokines and glaucoma.
5.Contents Determination of Nine Components in Guizhi Decoction and Comparison of Different Decoction Methods
Pengfei YUAN ; Huan LIU ; Mingying SHANG ; Feng XU ; Guangxue LIU ; Shaoqing CAI ; Lanfang LI ; Tingliang JIANG
China Pharmacy 2016;(6):801-804,805
OBJECTIVE:To establish a method for the contents determination of 9 components in Guizhi decoction,and com-pare the effects of traditional decoction method and the extracting machine decoction method on these contents in Guizhi decoction. METHODS:HPLC was performed on the column of Agilent Zorbax SB-C18 with mobile phaseof acetonitrile- 0.1% phosphoric ac-id(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 230 nm,254 nm and 280 nm,the column tempera-ture was 25℃,and the injection volume was 10μl. RESULTS:The linear range was 0.410 2-210.0μg/ml for gallic acid(r=0.999 9), 0.994 0-254.5μg/ml for albiflorin(r=0.999 9),1.636 0-1 675.0μg/ml for paeoniflorin(r=0.999 9),0.988 3-506.0μg/ml for liquiri-tin(r=0.999 6),0.987 3-31.59 μg/ml for coumarin(r=0.999 5),0.486 8-124.6 μg/ml for cinnamic acid(r=0.999 5),2.458 0-314.6μg/ml for cinnamaldehyde(r=0.999 5),0.034 3-1.096 μg/ml for 2-methoxy cinnamaldehyde(r=0.999 8),and 1.711 0-219.0 μg/ml for glycyrrdhizic acid (r=0.999 7);RSDs of precision,stability and reproducibility tests were lower than 5%,recoveries were 93.56%-103.19%(RSD=4.00%,n=9)、101.51%-107.32%(RSD=2.21%,n=9)、95.08%-103.76%(RSD=2.87%,n=9)、100.82%-105.73%(RSD=1.85%,n=9)、85.08%-89.12%(RSD=1.40%,n=9)、92.31%-99.12%(RSD=2.71%,n=9)、99.17%-102.32%(RSD=1.24%,n=9)、100.15%-103.98%(RSD=1.18%,n=9)、99.93%-102.61%(RSD=1.03%,n=9). The content of total effective components from the extracting machine decoction method was 4 565μg/g,that from the traditional decoc-tion method was 2 742 μg/g.CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of 9 componentsin Guizhi decoction. The contents of gallic acid,albiflorin and 2-methoxy cinnamaldehyde are first re-ported. The total effective components from the extracting machine decoction method are higher than that from the traditional decoc-tion method.
6.Subcellular localization of macrophage migration inhibitory factor and its diverse biological functions in tumors
Huan ZHANG ; Chen YANG ; Shuang SHANG ; Xiao-xi LV ; Fang HUA
Acta Pharmaceutica Sinica 2023;57(8):2085-2097
Macrophage migration inhibitory factor (MIF) is an enzyme-active pleiotropic cytokine that is expressed in various immune cells and tumor cells. MIF plays diverse roles in inflammation and tumor progression. It acts as a cytokine involved in immune response and inflammatory lesions. Additionally, MIF is closely associated with tumor proliferation, metastasis, and other tumor hallmarks, exerting a multifaceted influence on tumor occurrence and progression. MIF not only functions by being secreted into the extracellular space as a cytokine but can also be localized within the cytoplasm and nucleus, exhibiting diverse biological functions. As MIF in promoting tumor progression becomes increasingly recognized, MIF-based therapeutic strategies have become a hot research topic in oncology. Here, we provide a comprehensive review of MIF with different subcellular localization about their pro-tumoral functions. A better understanding of MIF in tumor biology will bring broader perspectives for the development of novel MIF targeting strategies and give promising direction for future tumor treatments.
7.Expression of zinc finger protein X-linked in childhood B lineage acute lymphoblastic leukemia.
Zhuo WANG ; Huan-Huan LIANG ; Ben-Shang LI ; Xiao-Hang HUANG ; Jing ZHANG ; Xiang WANG ; Li-Xia DING ; Hua JIANG
Chinese Journal of Contemporary Pediatrics 2013;15(7):509-513
OBJECTIVETo study the expression of zinc finger protein X-linked (ZFX) in bone marrow mononuclear cells (BMMCs) of children with B lineage acute lymphoblastic leukemia (B-ALL) and its relationship with prognosis.
METHODSThe expression of ZFX in human leukemia cell lines (REH, HL-60, NB(4) and K562) was measured by Western blot. ZFX gene was cloned by PCR from one patient and DNA sequencing technology was used to confirm it. Real-time PCR was used for detecting ZFX mRNA expression in the BMMCs of 82 children with newly-diagnosed B-ALL, 24 children with complete remission (CR) after induction therapy and 64 control children (fracture or congenital heart disease patients). According to the presence of bone marrow or central nervous system relapse during a follow-up of 3 years, the patients were identified as having a good or poor prognosis. Their ZFX mRNA levels in BMMCs at diagnosis were compared.
RESULTSZFX protein was expressed in human leukemia cell lines REH, HL-60, NB(4) and K562. ZFX mRNA expression was significantly higher in the newly-diagnosed ALL group than in the control group (P < 0.01). ZFX mRNA expression in the ALL CR group was significantly reduced compared with the newly-diagnosed ALL group (P < 0.01). Children with a poor prognosis had significantly higher ZFX mRNA levels at diagnosis than those with a good prognosis (P < 0.05).
CONCLUSIONSZFX is over-expressed in children with B-ALL and its levels are higher in those with a poor prognosis than those with a good prognosis, which suggests that ZFX is important in the prognosis evaluation of B-ALL.
Adolescent ; Cell Line, Tumor ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Kruppel-Like Transcription Factors ; analysis ; genetics ; physiology ; Male ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Prognosis ; Real-Time Polymerase Chain Reaction
8.Clinical features of antiviral therapy-induced thyroid disease in patients with chronic hepatitis C.
Jun-Ping LIU ; Huan-Rong HOU ; Yi KANG ; Jia SHANG ; Yong-Ge CAO ; Shou-Qin LIANG ; Xiu JIN
Chinese Journal of Hepatology 2013;21(4):257-260
OBJECTIVETo investigate the clinical features of thyroid disease occurring in response to antiviral therapy in patients with chronic hepatitis C (CHC).
METHODSEighty-two patients diagnosed with CHC were recruited for study from our hospital between 2009 and 2010. All patients were given a 48-week course of antiviral combination therapy with pegylated-interferon (Peg-IFN; 180 mug qw ih) and ribavirin (RBV; 15 mg/kg bw). Patient sera was collected prior to treatment (baseline), at treatment weeks 24 and 48, and post-treatment week 24, and used to detect changes in levels of thyroid function markers, thyroid-specific and other autoantibodies, complement factors, and immunoglobulins (Igs). Differential expression of biomarkers was assessed between patients who developed thyroid disorder and those who did not.
RESULTSAt treatment week 48, 13.4% (11/82) of cases developed hypothyroidism, 3.7% (3/82) developed hyperthyroidism, 20.7% (17/82) tested positive for thyroglobulin antibody, and 22.0% (18/82) tested positive for thyroid peroxidase antibody. The patients who did not develop thyroid disease had significantly higher post-treatment levels (vs. baseline) of IgG (14.84 +/- 2.61 vs. 12.95 +/- 3.32 g/L, F = 10.458, P = 0.002) and C4 (0.26 +/- 0.09 vs. 0.22 +/- 0.08 g/L, F = 6.835, P = 0.011) and significantly lower IgM (0.86 +/- 0.48 vs. 1.00 +/- 0.42 g/L, F = 9.106, P = 0.003). The patients who developed thyroid disease showed no significant differences in the baseline and post-treatment levels of IgG, C4, or IgM. When the two groups of patients who did or did not develop thyroid disease were compared, there was no difference in the amount of patients who achieved sustained virological response.
CONCLUSIONAntiviral-induced thyroid disease in patients with refractory hepatitis C manifests as clinically-detectable abnormalities in serum levels of thyroid autoantibody and markers of hypothyroidism. Levels of other autoantibodies and Igs do not correlate with the development of thyroid disease in these patients, and thyroid disease does not appear to affect the efficacy of Peg-IFN + RBV antiviral therapy.
Antiviral Agents ; therapeutic use ; Drug Therapy, Combination ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Polyethylene Glycols ; therapeutic use ; Ribavirin ; therapeutic use ; Thyroid Diseases ; chemically induced
9.Construction and identification of recombinant human platelet-derived growth factor-B adenoviral vector and transfection into periodontal ligament stem cells.
Shu-huan SHANG ; Yu-feng ZHANG ; Bin SHI ; Xiang-rong CHENG
Chinese Journal of Stomatology 2008;43(10):584-588
OBJECTIVETo construct a recombinant human platelet-derived growth factor-B (PDGF-B) adenoviral vector and to transfect it into human periodontal ligament stem cells (PDLSC).
METHODSThe recombinant plasmid pAd-PDGF-B was constructed by homologous recombination and confirmed by restriction endonucleases digestion. Recombinant adenovirus was packaged in HEK293 cells. PDLSC were transfected with recombinant adenovirus and PDGF-B expression was confirmed. Expression of collagen type I gene was determined by quantitative analysis of the products of RT-PCR. The cell proliferation was determined with MTT colorimetric assay.
RESULTSThe recombinant plasmid pAd-PDGF-B was confirmed by restriction endonucleases digestion. EGFP expression was observed on the third day after transfecting, and the expression of PDGF-B was detected. Immunohistochemical methods revealed that PDGF-B was expressed in PDLSC. Levels of expression of collagen type I gene were increased significantly by transfer of the exogenous PDGF-B gene to PDLSC. At the same time, findings indicated that Ad-PDGF-B stimulated PDLSC proliferation. MTT assay indicated the absorbance of PDLSC by stimulating with Ad-PDGF-B was (0.68 +/- 0.02), P < 0.01.
CONCLUSIONSUsing the AdEasy system, the human PDGF-B recombinant adenovirus can be rapidly obtained. These results indicate that recombinant adenoviruses encoding PDGF-B transgenes could modulate proliferative activity of PDLSC, enhance the high expression of collagen type I and lay the foundation for periodontal tissue regeneration and dental implant gene therapy.
Adenoviridae ; genetics ; Cells, Cultured ; Genetic Vectors ; Humans ; Periodontal Ligament ; cytology ; Proto-Oncogene Proteins c-sis ; genetics ; Stem Cells ; Transfection
10.Preliminary molecular analysis of bacterial composition in periapical lesions with primary endodontic infections of deciduous teeth.
Jia-jian SHANG ; Qiu-bo YANG ; Huan-ying ZHAO ; Shuang CAI ; Yan ZHOU ; Zheng SUN
Chinese Medical Journal 2013;126(16):3112-3117
BACKGROUNDThe bacterial composition of periapical lesions in deciduous teeth has not been well documented. This study was designed to explore the bacterial compositions, especially the dominant bacteria in periapical lesions using 16S rRNA sequencing.
METHODSTissue samples were collected from 11 periapical lesions in deciduous teeth with primary endodontic infections. DNA was extracted from each sample and analyzed using 16S rRNA cloning and sequencing for the identification of bacteria.
RESULTSAll DNA samples were positive for 16S rRNA gene PCR. One hundred and fifty-one phylotypes from 810 clones were identified to eight phyla, and each sample contained an average of 25.9 phylotypes. In addition, 59 phylotypes were detected in more than two samples, and Fusobacterium (F.) nucleatum (8/11), Dialister (D.) invisus (8/11), Campylobacter (C.) gracilis (7/11), Escherichia (E.) coli DH1 (6/11), Aggregatibacter (A.) segnis (6/11), and Streptococcus (S.) mitis (6/11) were the most prevalent species. Furthermore, 45 as-yet-uncultivated phylotypes were also identified.
CONCLUSIONSChronic periapical lesions in deciduous teeth contained polymicrobial infections. F. nucleatum, D. invisus, C. gracilis, E. coli DH1, A. segnis, and S. mitis were the most prevalent species detected by 16S rRNA sequencing.
Bacteria ; classification ; genetics ; isolation & purification ; Bacterial Infections ; microbiology ; Child ; Child, Preschool ; Female ; Humans ; Male ; Periapical Tissue ; microbiology ; RNA, Ribosomal, 16S ; genetics ; Tooth, Deciduous ; microbiology