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

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.

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.

Objective To observe the influence of hIL-10-MSCs on apoptosis of rats subject to discordant liver xenotransplantation. Methods The model of guinea pig to rat discordant liver xenotransplantation was set up. The orthotopic liver transplantation model was established by using modified two-cuff technique. Following groups were designed: control group, MSCs group and hIL-10MSCs group. Before and after operation, the recipients in control group were injected with normal sodium (2 ml) and decaesadril (0.5 ml) ;Those in MSCs group were injected with MSCs (4.0× 106/ml)and decaesadril (0.5 ml); Those in hIL-10-MSCs group were injected with hIL-10-MSCs (4.0 × 106/ ml)and decaesadril (0.5 ml). Twelve h after operation, the livers of recipient were observed by HE staining and electron microscope, and apoptosis of the livers was detected by using TUNEL. The expression levels of hIL-10, caspase-3, Fas and FasL were assayed by Western blot or immunohistochemistry. Results As compared with control group and MSCs group, the pathological changes was alleviated, the expression of IL-10 was significantly increased, the expression of FasL was significantly reduced in hIL-10-MSCs group. The positive area size of Fas and Caspase-3 in hIL-10MSCs group was 11.5 % and 25.1 %, respectively, significantly smaller than those in control group (35.3 % and 70.8 % respectively, P＜0.05. Apoptosis index in hIL-10-MSCs group was 32.5 %,which was significantly lower than in control group (74.1%) and MSCs group (50. 3 % ). ConclusionhIL-10-MSCs can notably decrease apoptosis of the transplanted liver probably by inhibiting theexpression of Fas/FasL.

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.

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 investigate the clinical value of procalcitonin (PCT) levels in bacteria identification in intensive care unit (ICU) patients with bloodstream infection.Methods There were 540 cases of patients with bloodstream infection in our ICU between December 2007 and December 2013.The PCT levels and bacteria were identified.The application effectiveness of PCT levels in the bacteria identification was studied.Results The G+ bacteria infection rate was 49.63％ (268/540),G-bacteria infection rate was 38.52％ (208/540),and the fungal infection rate was 11.85％ (64/540).The patients of G-bacteria had significant difference with G + bacteria and fungal infection (P ＜ 0.05).The PCT average and positive rate of G-bacteria were significantly higher than G + bacteria and fungi group (P ＜ 0.05),respectively.G+ bacteria and fungi infection did not have significant difference (P ＞ 0.05).When PCT ＞ 2.04 ng/ml,the sensitivity and specificity that applying serum PCT level to identify the between G-and G+ bacteria were 82.18％ and 76.09％,respectively.When PCT ＞3.16 ng/ml,the sensitivity and specificity that applying serum PCT level to identify the between G-and fungus bacteria were 59.42％ and 65.73％,respectively.Conclusions The identification between G-bacteria and G + bacteria,fungi with applying PCT level in bloodstream infections had high accuracy.When the PCT levels was greater than 2.04 ng/ml,the occurrence of G-bacteria was greater risk of infection.The accuracy of PCT level identifying the G + bacteria and fungi was poor.

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.