1.Regulation of Jinxin Oral Liquid for the expression of negative regulatory factor of TLR3 signaling pathway SOCS1 in RSV infected BALB/c mice.
Zheng-Guang CHEN ; Shou-Chuan WANG ; Jian-Ya XU ; Qi-Gang DAI
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(12):1499-1506
OBJECTIVETo investigate the regulation trend of Jinxin Oral Liquid (JXOL) on the expression of negative regulatory factor of TLR3 signaling pathway SOCS1 in the lung tissue of RSV infected BALB/c mice at different time points.
METHODSTotally 75 BALB/c mice were randomly divided into 5 groups, i.e., the normal control group, the model group, the ribavirin group, the high dose JXOL group, and the equivalent dose JXOL group, 15 in each group. Each group had 3 intervention ways (I, II, and III) with 5 mice treated in each group. BALB/c mice were nasally infected with respiratory syncytial virus (RSV), and treated by different intervention ways. After intervention, mice were killed and their lung tissues were sampled, mRNA expression levels of RSV-M, SOCS1, and IFN-β were detected by Real time PCR. The expression of SOCSl at the protein level was detected by Western blot.
RESULTSCompared with the normal control group, the mRNA expression level of SOCS1 and IFN-β, and the protein expression level of SOCS1 increased significantly in the model group intervened by intervention I and II (all P < 0.01), but the mRNA expression level of IFN-β decreased significantly in model group intervened by intervention III (P < 0.01). Compared with the model group, the mRNA expression level of RSV-M all significantly decreased in the high dose JXOL group and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01). The mRNA expression level of SOCS1 significantly decreased in the high dose JXOL group intervened by intervention I and III and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01). The mRNA expression level of IFN-β significantly decreased in the high dose JXOL group intervened by intervention I and II and the equivalent dose JXOL group intervened by intervention I (all P < 0.01), while it significantly increased in the high dose JXOL group intervened by intervention III and the equivalent dose JXOL group intervened by intervention III (all P < 0.01). The protein expression level of SOCS1 significantly decreased in the high dose JXOL group intervened by intervention I and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01), while it significantly increased in the high dose JXOL group intervened by intervention III (all P < 0.01). Compared with the high dose JXOL group, the mRNA expression level of RSV-M decreased significantly in the equivalent dose JXOL group intervened by intervention I and II (P < 0.01). The mRNA expression level of SOCS1 and IFN-β decreased significantly in the equivalent dose JXOL group intervened by intervention I (P < 0.01), but the mRNA expression level of IFN-β increased significantly in the equivalent dose JXOL group intervened by intervention II and III (all P < 0.01). The protein expression level of SOCS1 decreased significantly in the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01).
CONCLUSIONSJXOL could inhibit the expression of SOCS1 in the lung tissue of RSV infected BALB/c mice at different time points. Its regulatory effect might be associated with promoting the expression of interferon type I and further fighting against RSV.
Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Lung ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; Respiratory Syncytial Virus Infections ; drug therapy ; metabolism ; Respiratory Syncytial Viruses ; Ribavirin ; Signal Transduction ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling Proteins ; metabolism ; Toll-Like Receptor 3 ; metabolism
2.Surgical treatment for chronic achilles tendon rupture and severe scarring.
Chuan-Xiu SUN ; Sheng-Wei HE ; Xu FANG ; Li-Dong MI ; Guang-Yu DU ; Xue-Gang SUN
China Journal of Orthopaedics and Traumatology 2015;28(4):354-356
OBJECTIVETo evaluate the clinical efficacy of autologous semitendinosus and gracilis tendon grafting with anchor repair for the treatment of chronic achilles tendon rupture and severe scarring.
METHODSFrom April 2010 to October 2012,26 patients with chronic achilles tendon rupture(with Myerson type III ) and severe scarring were treated with autologous semitendinosus and gracilis tendon grafting with anchor repair. There were 19 males and 7 females,with an average age of 32 years old (ranged, 22 to 47 years). The time from injury to surgery was from 3 to 12 months (7 months on average). The plantar flexion strength of all injuried feet attenuated and single heel rise test were positive in 26 cases before operation. Plaster immobilization and routine rehabilitation therapy were performed after operation. Clinical effects were evaluated by Arner-lindholm criterion and complications were observed after operation.
RESULTSAll the patients were followed up from 12 to 24 months with a mean of 16 months. No complications such as achilles tendon re-rupture, wound infection, etc were found during follow-up period. According to the Arner-Lindholm standard, 15 cases got excellent results and 11 good.
CONCLUSIONUsing autologous semitendinosus and gracilis tendon grafts with anchor repair to treat chronic achilles tendon rupture and severe scarring is a perfect surgical procedure.
Achilles Tendon ; injuries ; surgery ; Adult ; Chronic Disease ; Cicatrix ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Rupture ; Young Adult
3.Molecular Genetic Analysis of One Sudden Unexplained Death in the Young by Whole Exome Sequencing.
Chun WANG ; Hui WANG ; Xin-shu XU ; Chuan-chao XU ; Xiao-ping LAI ; Rui CHEN ; Han-guang LIN ; Sheng-yuan QIU
Journal of Forensic Medicine 2015;31(6):436-444
OBJECTIVE:
To find the mutation of disease-causing genes of sudden unexplained death syndrome (SUDS) in the young by whole exome sequencing in one case.
METHODS:
One SUDS case was found no obvious fatal pathological changes after conventional autopsy and pathological examination. The whole exome sequencing was performed with the Ion Torrent PGM™ System with hg19 as reference sequence for sequencing data. The functions of mutations were analyzed by PhyloP, PolyPhen2 and SIFT. A three-step bioinformatics filtering procedure was carried out to identify possible significative single nucleotide variation (SNV), which was missense mutation with allele frequency < 1% of myocardial cell.
RESULTS:
Four rare suspicious pathogenic SNV were identified. Combined with the analysis of conventional autopsy and pathological examination, the mutation MYOM2 (8_2054058_G/A) was assessed as high-risk deleterious mutation by PolyPhen2 and SIFT, respectively.
CONCLUSION
Based on the second generation sequencing technology, analysis of whole exome sequencing can be a new method for the death cause investigation of SUDS. The gene MYOM2 is a new candidate SUDS pathogenic gene for mechanism research.
Autopsy
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Brugada Syndrome/genetics*
;
Cause of Death
;
DNA Mutational Analysis/methods*
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Death, Sudden/etiology*
;
Exome
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Gene Frequency
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Genetic Testing/methods*
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High-Throughput Nucleotide Sequencing/methods*
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Humans
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Molecular Biology
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Molecular Diagnostic Techniques/methods*
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Molecular Sequence Data
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Mutation
4.Expression of HBcAg in eukaryotic cells by retroviral vector mediated gene transfer.
Chuan-lin DING ; Kun YAO ; Tian-tai ZHANG ; Jiang-ying XU ; Lin XU ; Guang-yong PENG
Chinese Journal of Experimental and Clinical Virology 2003;17(1):81-84
BACKGROUNDTo construct recombinant retroviral vector expressing HBcAg in eukaryotic cells.
METHODSThe HBV core gene fragment was amplified by using PCR from pADR which contains complement nucleotide sequence of HBV subtype adr and cloned into retroviral expression plasmid pLXSN, then transfected into packing cell (PT67) with lipofec AMINE. After 2-3 weeks selection with G418, large colonies were isolated and transferred to individual plates. Virus-containing medium was collected and used to infect NIH3T3, EL4 and mouse bone marrow derived dendritic cells(DC). DNA was extracted from infected cells and tested by PCR. Indirect immunofluorescence and FACS were used to detect the expression of HBcAg. Cell mediated immunity of immunized C57BL/6 mice with transduced DC was analyzed.
RESULTSThe insertion of HBV core gene fragment in the recombinant retroviral plasmid was confirmed by PCR as well as enzyme digestion with EcoR1 and BamH2. The viral titer reached 3 x 10(5) CFU/ml. The result of PCR showed that the HBV core gene had been integrated into the genome of infected NIH3T3 cells. Indirect immunofluorescence and FACS analysis showed that HBcAg had been expressed in these cells. HBcAg specific CTL responses could be generated in mice immunized with retrovirus transduced DC.
CONCLUSIONSHBV core gene had been integrated into eukaryotic cells with retroviral vector and target gene had been expressed efficiently. These results may have some impact on gene therapy of chronic hepatitis B.
3T3 Cells ; Animals ; Cloning, Molecular ; Dendritic Cells ; metabolism ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; Humans ; Mice ; Recombination, Genetic ; genetics ; Retroviridae ; genetics ; Transfection
5.High intensity focused ultrasound for liver cancer.
Chuan-xing LI ; Guo-liang XU ; Jian-jun LI ; Guang-yu LUO
Chinese Journal of Oncology 2003;25(1):94-96
OBJECTIVETo evaluate the effect of high intensity focused ultrasound (HIFU) for liver cancer.
METHODSHIFU treatment was performed in 44 liver cancer patients under general anesthesia and ultrasound positioning. Before and after HIFU treatment, the clinical symptoms, liver functional tests, AFP and MRI were evaluated.
RESULTSAfter HIFU treatment, the remission rate of clinical symptoms was 87.5% (28/32), with the scanty ascites in 3 patients disappeared. ALT (79.73 +/- 12.31 U/L) and AST (103.47 +/- 24.55 U/L) before HIFU were reduced to normal in 84.6% (22/26) and 73.5% (25/34) patients. AFP in 64.3% (18/28) patients decreased > or = 50% of the original value. After HIFU, MRI showed coagulative necrosis and blood supply reduction or disappearance of tumor in the target region.
CONCLUSIONHIFU treatment pocesses good effect on liver cancer, which will offer a considerable weight in noninvasive local treatment of hepatic tumor.
Adult ; Aged ; Female ; Humans ; Liver Neoplasms ; therapy ; Male ; Middle Aged ; Ultrasonic Therapy ; Ultrasound, High-Intensity Focused, Transrectal
6.Classification and morphology of jugular bulb and its clinical significance
Guang-Yong TIAN ; Da-Chuan XU ; De-Liang HUANG ; Lu-Jun HAN ; Zhi-Qiang PENG ; Ze-Yu LI ; Xiao-Tian SHI
Chinese Journal of Neuromedicine 2008;7(5):483-486,494
Objective To observe the anatomic and imaging morphology ofjugnlar bulb and its relationship with the surrounding structures, and to investigate the classification ofjugnlar bulb and its clinical significance. Methods We dissected 30 human temporal bones and studied multi-slice spiral CT imaging data of temporal bone of 120 cases and blood vessel cast mould specimen of the jugular bulb of 6 cases, to observe the morphology of jugnlar bulb and its spatial relationship with the surrounding structures. We made an imagined sagittal plane on the medial well of the tympanic cavity, with a horizontal tangent line of the proximal wall of the tympanic cavity and a vertical tangent line of the posterior wall of the tympanic cavity as coordinate axes (X axis and Y axis), respectively, so the 4 quadrants ( Ⅰ , Ⅱ, Ⅳ, Ⅳ) were formed. The jugular bulb was classified intro 4 types according to the quadrant where its top was projected and subtyped according to its position on the inner or outer side of the plane. The operation via mastoid approach was simulated on specimen to observe the effect of jugnlar bulb on the operation route. Results Some jugular bulbs were flat type and others were prominent types. The classification in the group of CT image: type Ⅰ , 11 case (9%);type Ⅱ, 63 cases (53%);type Ⅲ, 25 cases (21%);type Ⅳ, 21cases (17%). The classification in the group of specimen: type Ⅰ, 1 case (3%);type Ⅱ, 11 cases (37%);type Ⅲ, 8 cases (27%);type Ⅳ, 10 cases (33%). Each type of the jugular bulb had different effects on the operative approach. Conclusions The classification method with the 4 quadrants is a simple and three-dimensional way to describe the position of the jugular bulb for imaging diagnosis or operative scheme design.
7.A microanatomic study of endoscope-assisted presigmoid-retrolabyrinthine keyhole approach to the petroclival region
Zhi-Qiang PENG ; Guang-Yong TIAN ; Yong-Yuan HE ; Da-Chuan XU
Chinese Journal of Neuromedicine 2009;8(8):810-812
Objective To provide an anatomical basis for endoscope-assisted presigmoid-retrolabyrinthine keyhole approach to the petroclival region. Methods Ten formalin-fixed adult cadaver heads were used in this study. A C-shaped postauricular skin incision about 6 cm in length was made to expose the anatomic structures, which were observed under microscope and endoscope. Results The distance from the anterior margin of the sigraoid sinus to the most posterior point of the posterior semicircular canal was 9.1±1.3 mm on the right and 9.6±1.8 mm on the left side. With appropriate adjustment of the endoscope and the angle of the microscope, the presigmoid-retrolabyrinthine keyhole approach allowed good exposure of the important structures in the petroclival region with also clear display of the cranial nerves and the adjacent blood vessels. Conclusion The facial and vestibulocochlear nerves and the posterior orifice of the internal auditory canal can be used as the landmarks for localization of the peroclival region and its surrounding structures.
8.A Novel Neutralizing epitope of Human cytomegalovirus glycoprotein M Screened by Phage Display
Benxu WANG ; Yu LIU ; Zhan LIU ; Yaping GAO ; Fang WANG ; Heping PAN ; Guang YANG ; Hua XU ; Beifen SHEN ; Chuan LIU ; Ningsheng SHAO
Progress in Biochemistry and Biophysics 2009;36(2):220-227
Human cytomegalovirus glycoprotein complex Ⅱ (gC Ⅱ ) consists of two glycoproteins, gM and gN. Although gC Ⅱ specific IgG purified from HCMV positive patient sera can neutralize HCMV, there has been no report on the generation of virus-neutralizing antibodies by immunizing with one epitope of gM. The epitope, termed MAD, was screened from random phage peptide library by subtractive strategy. The peptide sequence of MAD was highly homologous with 32~38 amino acids of HCMV gM. Mice immunized with MAD coupled with keyhole limpet hemocyanin (KLH) could produce specific antibodies against MAD, and the antibodies obtained could bind not only native HCMV particles, but also the recombinant gM30~78 peptide. ELISA analysis results showed that MAD could specifically bind HCMV-positive human serum samples. Virus-neutralizing assay results demonstrated that the antibodies against MAD could inhibit HCMV strain AD169 entering the human embryonic lung cells. The results suggested that MAD could be used as a new potential protective antigen in the development of HCMV vaccine.
9.Development of cell-penetrating peptides as vectors for drug delivery.
Jin REN ; Chuan-Guang QIN ; Chun-Lan XU ; Qiu-Yu WANG ; Xiao-Jia ZUO
Acta Pharmaceutica Sinica 2010;45(1):17-25
Biomacromolecules play an important role in the treatment of many diseases, but as a result of cell membrane serving as the natural barriers, only the small molecular compounds whose molecular weights are smaller than 600 Da can get through cell membrane and enter the cell. In recent years, some short peptides (the length less than 30 amino acids) are found to have the cell-penetrating function, called cell-penetrating peptides (CPPs). They are able to effectively translocate segments of protein, polypeptides, nucleic acid into the cells of many mammal animals with many methods. They have high transduction efficiency and will not lead to cell damage. So, the discovery of CPPs has a very good applicable prospect in such research fields as cell-biology, gene-therapy, drug transduction in vivo, evaluation of clinical medicine and medical immunology. This paper reviews the types and characteristics of CPPs, internalization mechanisms, applications, and their existing problems.
Absorption
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drug effects
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Amino Acid Sequence
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Animals
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Cell Membrane Permeability
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Cell-Penetrating Peptides
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classification
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pharmacology
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physiology
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Drug Carriers
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Endocytosis
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physiology
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Humans
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Protein Transport
10.Clinical application of real time-polymerase chain reaction in determining cytomegalovirus viral DNA load in renal transplant recipients.
Chuan-Bao ZHANG ; Hui-Ying LAI ; Hong-Tao XU ; Da-Guang WANG ; Fei XIAO
Chinese Medical Journal 2012;125(19):3575-3577
BACKGROUNDCytomegalovirus (CMV) remains a significant clinical problem among immunosuppressed renal transplant patients. Quantitative PCR assays have become the most common methods in the determination of CMV infections in transplant patients. This study was to determine the relationship between CMV infection and the acute rejection of the transplanted kidney.
METHODSPlasma samples from 77 renal transplant patients that were pre-transplant negative for CMV infection were tested using real-time quantitative PCR and CMV gene-specific primers. The detected viral loads were retrospectively compared with the acute rejection rate and the chronic or mild rejection rates of the renal transplant.
RESULTSCMV-DNA was detected in 29 of 77 recipients, yielding a positive rate of detection of 37.7% for this procedure. Twelve of the 21 recipients (57.1%) who suffered acute rejection had positive CMV-DNA. Among the 56 recipients suffered from chronic or mild rejection, 17 (30.4%) had positive CMV-DNA plasma. Moreover, of the 29 recipients who had detectable CMV-DNA after transplant, 12 (41.4%) suffered from acute rejection; of the 48 recipients with undetectable CMV-DNA, only nine (18.8%) developed acute rejection. Post-transplant patients with acute rejection had a higher rate (57.1% vs. 30.4%, P = 0.03) of post-transplant CMV infection than those with chronic or mild rejection.
CONCLUSIONCMV infection is a risk factor of acute renal transplant rejection and CMV infection should be prevented and treated in renal transplant recipients.
Adult ; Cytomegalovirus Infections ; diagnosis ; genetics ; DNA, Viral ; genetics ; Female ; Humans ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction ; methods ; Viral Load ; Young Adult