Objective To establish hepatocellular carcinoma(HCC) cell lines which olig-expressed IGF1R gene stably.Methods An eukaryotic expressing vector pSUPER-IGF1R-siRNA that could block IGF1R expressing was transferred into hepatocellular carcinoma cell lines SMMC7721 and Hep3B with Lipofectamine 2000 reagents.After transferred,cells were selected with G418 to obtain positive clones.The expressions of IGF1R,cyclin D1 and cyclin B1 were detected by RT-PCR and Western-blot.Cell growth curve were painted.Results Two cell lines clones were screened olig-expressing IGF1R gene stably.The experimental cell lines grew more slowly than control cell lines and the expression of cyclin D1 decreased(P

Objective To study the effect of Fufangbiejiaruanganpian (FFBJRGP) on deposition of collagens around hepatic Disse space in experimental alcohol liver fibrosis (ALF). Methods The ALF rat model was induced by feeding a mixture of alcoholic liquid which included alcohol, vegetable oil and pyrazole, combined with a high fatty food. Then, all of the ALF rats were treated with FFBJRGP in high, moderate, and low dose. At the time of termination of the treatment, HYP level in liver tissue, collagen Ⅳ and LN levels in the serum and pathological changes were studied by Mallory stain and sirius red stain. Results The levels of HYP in hepatic tissues, collagen Ⅳ and LN in sera are increased remarkably in ALF rats, while all these indexes were reduced after moderate or low-dose FFBJRGP treatment (P

This article summarized the technique research and application status of near-infrared spectroscopy (NIRS), in order to explore hypoxia,qi-deficiency and their connections with typical symptoms. It also discussed NIRS acquired oxygen saturation (SaO2) and its relevant means and methods for information quantification ofqi-deficiency. The accurate obtained SaO2 value was applied in the association with the strength of chest distress in the subjective sensation due toqi-deficiency. According to current data, the normal value,qi-deficiency degree and uncomfortable degree of subjective sensation among patients of different altitudes and ages were summarized. It can be applied to assist TCM physicians in the quick quantification of qi-deficiency.

Cucumber mosaic virus was detected from infected Basella rubra L. with the indirect enzyme-linked immunosorbent assay. Total RNA was extracted from infected leaves and the cDNAs of coat protein gene of CMV-Ba were obtained by RT-PCR. The amplified cDNA fragments were then cloned into pMD 18-T vector and sequenced,the result showed that the CP gene was 657 nucleotides in length. This sequence was aligned with the obtained CP gene and some CMV strains or isolates of subgroup Ⅰ and subgroup Ⅱ in GenBank using DNA MAN software. The results showed that CMV-Ba shared 90.9%~93.8% and 76.1%~76.9% identity with the known CP genes of subgroup Ⅰ and Ⅱ respectively in nucleotide level,on the other hand,amino acids deduced from CMV-Ba CP gene shared 92.7%~97.7% and 72.4%~78.1% identity with the known CP protein of subgroup Ⅰ and Ⅱ,respectively. This suggested that CMV-Ba CP gene belongs to CMV subgroupⅠ.

Objective To observe the influence of hIL-10-MSCs on rejection of rat discordant liver xenotransplantation. Methods The model of cavia to rat discordant liver xenotransplantation was established. The rats were randomized into the control group, MSCs group, and hIL-10-MSCs group.The survival time and liver function of each recipient were observed and expression levels of E-Selectin, LFA-1, VCAM-1, and NF-κB determined by RT-PCR and ELISA 24 h after transplantation.Results The survival time was prolonged, liver function improved, and expressions levels of E-Selectin, LFA-1, VCAM-1 and NF-κB were significantly decreased in the hIL-10-MSCs group (P＜0.05).Conclusion hIL-10-MSCs are able to protect transplanted liver and decrease the expression of E-Selectin, LFA-1, VCAM-1, and NF-κB in the liver.

BACKGROUND:Extradural cortical stimulation combines the advantages of repetitive transcranial magnetic stimulation, transcranial direct current stimulation, subdural cortical stimulation and deep brain stimulation, which can significantly improve motor and language function after stroke. OBJECTIVE:To review the theoretical research and clinical application of extradural cortical stimulation for stroke recovery. METHODS:An online retrieval of PubMed database and CNKI database between January 1995 and April 2014 was performed for articles on theoretical research and clinical application of extradural cortical stimulation for stroke recovery, with the key words of“cortical stimulation, extradural motor cortex stimulation, extradural cortical implants, extradural cortical stimulation, stroke, rehabilitation”in English and Chinese. RESULTS AND CONCLUSION:Because of implantable cortical stimulation, the advantage of extradural cortical stimulation is its minimal invasiveness, high accuracy and transdural contact with the brain. For lack of effective treatment for the chronic phase of stroke patients with motor and language dysfunction, extradural cortical stimulation may be a new therapeutic method. Motor and language functional improvement must derive from reactivation of plasticity, local enhancement of perilesional areas, enhancement of network function and inter-hemispheric balance function, and amplification of sensory input.

Objective To study the effects of lentiviral vector of microRNA targeting IGF1R gene on growth of liver cancer cells.Methods The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pPRIME vector,named pPRIME-IGFIR-miR30-shRNA.The viruses were propagated on 293T cells.Viruses were purified by CsCI gradient according to standard techniques,and functional PFU titers were determined by plaque assay on 293 cells.The effect of pPRIME-IGFIR-miR30-shRNA on IGFIR expression of Hep3B、SMMC7721 cells was detected by RT-PCR and Western blot.The antitumor potential of PRIME-IGFIR-miR30-shRNA to Hep3B、SMMC7721 cells was evaluated by CCK-8 assay and Tunel.Results PRIME-IGFIR-miR30-shRNA was constructed successfully.Functional PFU titers of pPRIME-IGFIR-miR30-shRNA were 4.58?109PFU/ml.PRIME-IGFIR-miR30-shRNA inhibited IGFIR expression and the proliferation of Hep3B、SMMC7721 cells.Conclusions PRIME-IGFIR-miR30-shRNA expressing IGFIR-siRNA can inhibit IGFIR expression and may be used for further investigation of gene therapy of liver cancer.

AIM: To explore the effect of pulmonary arterial smooth muscle cell (PASMC) apoptosis on the reversal of hypoxic pulmonary arterial remodeling during reoxygenation and its possible mechanism.METHODS: Male SD rats (n=24) were randomly divided into normoxia for 4 weeks group, hypoxia for 4 weeks group, reoxygenation for 1 week after hypoxia for 4 weeks group and reoxygenation for 6 weeks after hypoxia for 4 weeks group.Right ventricular systolic pressure (RVSP), right ventricular hypertrophy index, pulmonary arterial medial thickness (MT) and medial area (MA) as well as autophagy and apoptosis in the pulmonary arterial medial layer were examined during hypoxia-reoxygenation.The rat primary PASMCs were divided into normoxia for 48 h group, hypoxia for 48 h group, reoxygenation for 24 h after hypoxia for 48 h group and normoxia for 72 h group to explore the changes of PASMC autophagy and apoptosis following hypoxia-reoxygenation.Finally, primary PASMCs were divided into normoxia for 72 h group, reoxygenation for 24 h after hypoxia for 48 h group and reoxygenation for 24 h after hypoxia for 48 h + chloroquine (inhibitor of autophagy) group to investigate the effect of PASMC autophagy during hypoxia on the apoptosis during reoxygenation.RESULTS: After hypoxia for 4 weeks, the RVSP, during right ventricular hypertrophy index, MT and MA increased significantly compared with normoxia group (P<0.05), and gradually decreased during reoxygenation.The expression of LC3 in the pulmonary arterial medial layer increased evidently after hypoxia and gradually reversed during reoxygenation.Moreover, the P62 and cleaved caspase-3 expression decreased after hypoxia compared with normoxia group, and increased markedly following reoxyge-nation.The expression of cleaved caspase-3/PARP in rat primary PASMCs decreased significantly under hypoxia (P<0.05), and increased evidently during reoxygenation.The expression of P62 and LC3-II decreased markedly under hypoxia (P<0.05).After inhibition of PASMC autophagy under hypoxia, the expression of cleaved caspase-3/PARP decreased remarkably during reoxygenation (P<0.05).CONCLUSION: The PASMC apoptosis participates in the reversal of hypoxic pulmonary arterial remodeling, and the PASMC autophagy under hypoxia might facilitate its apoptosis during reoxygenation.

Objective · To design and build a high-throughput sequencing approach based on targeted panel sequencing (TPS) using for the primary immunodeficiency disease (PID) diagnosis. Methods · By reviewing the literature and querying the relevant databases to determine the known diseasecausing genes of PID, capture probes using for the TPS were designed and customized for all exons and flanking sequences of these genes. A child suspected with PID was diagnosed by the customized TPS. Results · The PID sequencing panel contains a total of 100 known pathogenic genes. The sequencing data of the patient has 16414298 reads. The average coverage depth is 157 X, 98.35% of the target region sequencing depth is greater than 20 X, and 99.97% of the target region sequencing depth is greater than 1 X. Finally, a heterozygous nonsense mutation was found in the exon 2 of the CXCR4 gene (c.1000C>T, p.Arg334*) in the child. The results of Sanger sequencing confirmed the variation in the child and showed that his parents were wildtype at the corresponding sites, indicating the mutation is de novo. Conclusion · This study established a high-throughput sequencing diagnostic approach for PID, with which a case of WHIM syndrome was successfully diagnosed.

Objective To observe the effects of hIL-10-MSCs on heterologous T cell prolifera-tion. Methods hlL-10 was cloned by RT-PCR and then hIL-10/pLOX-cwGFPs was construct. The lentivirus was transfected into cavy MSCs and cell culture supernatant was tested for IL-10 protein by ELISA assay. The effects of hIL-10-MSCs on heterologous T cell proliferation were determined by CCK-8. The effect of peripheral blood mononuclearcell secretion IFN-γ was tested by EIASA. Results hIL-10/pLOX-cwGFP were constructed successfully and hIL-10-MSCs cell culture supernatant of IL-10 protein were increased obviously, hIL-10-MSCs could inhibit heterologous T cell proliferation sig-nificantly(P<0.05) but could not induce T cell proliferation (P<0.05). Adding IL-2 could reverse this inhibition(P<0.05), hIL-10-MSCs cell culture supernatant could inhibit peripheral blood mono-nuclearcell secretion IFN-γ. Conclusion hIL-10-MSCs may play an important role in significant im-munosuppression of heterologous T cell proliferation and suppression of secretion IFN-γ by peripheral blood mononuclear cells.