Mechanism of Wen Shen Xuan Bi Tang in treatment of osteoporosis based on network pharmacology and animal experimental verification / 中国药理学通报

Aim To predict and validate the mechanism of wenshen xuanbi tang(WSXBT) in treatment of osteoporosis (OP) based on network pharmacology, molecular docking techniques and in vivo experimental techniques. Methods Network pharmacology was used to screen the key ingredients and core targets of WSXBT for the treatment of osteoporosis. Metascape database was used for gene ontology (GO) biological process enrichment analysis and kyoto encyclopedia of genes and genomes (KEGG) signaling pathway enrichment analysis of core targets. AutoDockTools 1. 5. 7 software was applied in molecular docking to simulate the binding activity of key active ingredients to core targets. To study the efficacy of WSXBT on rats with osteoporosis and to verify the related targets and pathways, rat models of osteoporosis were established by excising the bilateral ovaries of rats. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum OPG, PINP and RANKL content. Biomechanical tester was applied to test the biomechanics of rat femurs. Micro-CT was applied to detect the femoral bone density. Then, Western blot was employed to measure the protein expression levels of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt). Results A total of 156 active ingredients of WSXBT were screened, involving 229 potential targets, 23 core targets and 145 signaling pathways. The molecular docking results showed that five key ingredients, including quercetin, kaempferol, naringenin, isobavachin and licochalcone a, possessed good binding ability to the core targets of PIK3R1 and AKT1. The results of in vivo experiments showed that WSXBT could significantly increase bone density, improve bone tissue microstructure, enhance femur biomechanics and increase PINP expression and OPG/RANKL ratio in rats with osteoporosis. Results of WB showed that WSXBT significantly increased p-PI3K/PI3K and p-Akt/Akt ratios. Conclusions WSXBT could improve bone mineral density in postmenopausal osteoporotic rats through PI3K/ Akt signaling pathway and increasing OPG/RANKL ratio.

bFGF and IGF-1 promotes differentiation of bone mesenchymal stem cells into cardiomyocyte-like cells and its related mechanisms / 中国药理学通报

Aim To elucidate the biological effects of insulin-like growth factor-1 ( IGF-1 ) and basic fibro-blast growth factor (bFGF) alone or in combination on the differentiation of bone mesenchymal stem cells (BMSCs) into cardiomyocytes (CMs), and to explore the mechanism in the differentiation process of BMSCs into CMs induced by IGF-1 or bFGF. Methods After four weeks of BMSCs induced by induction differentiation medium ( with or without LY294002) containing IGF-1 and/or bFGF, the expression levels of proteins associated with the cardiomyogenic phenotype in BMSCs were detected via immunocytochemistry, immuno-fluorescence staining, and Western blot. Meanwhile, the expression levels of pathway related proteins were detected by Western blot. The cell ultrastructure was observed by transmission electron microscopy (TEM) . The expression levels of myocardial specific genes were measured via RT-qPCR. Results Compared with the control group, the expression levels of myocardial specific proteins and genes significantly increased in the IGF-1, bFGF and combination groups and were the highest in the coinduction group. The TEM of the conduction group showed parallel myofilaments, mitochondria, endoplasmic reticulum and so on. The additon of LY294002 decreased the expression levels of myocardial specific proteins, genes and phosphorylation Akt. Conclusions The effect of IGF-1 combined with bFGF on the differentiation of BMSCs into CMs is markedly better than that induced by IGF-1 or bFGF a-lone, and the differentiation process may depend on the PI3K/Akt signaling pathway.

Role of fibroblast growth factor 2 and tanshinoneⅡA in the differentiation of rat bone marrow mesenchymal stem cells into cardiomyocyte￣like cells / 解剖学报

Objective To investigate the effects of fibroblast growth factor 2 (FGF-2) combined with tanshinoneⅡA on differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into cardiomyocyte-like cells. Methods BMSCs were isolated and cultured. The cultured cells were identified by flow cytometry, BMSCs were divided into experimental control group, FGF-2 group, tanshinoneⅡA group and the combined induction group. The activity and value of BMSCs were detected by MTT. The Real-time PCR method was used to detect BMSCs. Expression of early myocardial transcription factors GATA-4 and Nkx2. 5; Immunocytochemical staining for detection of connexin43(Cx43)and cardiac troponin-Ⅰ(cTnI); Immunofluorescence staining for detection of desmin and Tm; The expression of desmin and tropomyosin(Tm)was detected by Western blotting. Results The cell activity and proliferation after induction were good. The expression of GATA-4 and Nkx2. 5 in the induction group were stronger than that in the experimental control group, and the difference was statistically significant (P<0. 05). The positive expression of Cx43 and cTnI in the induction group increased significantly. The markers of the combined group were the most obvious, and the difference was statistically significant (P<0. 05). The expression of Myo-specific protein desmin and Tm in the combined induction group increased significantly, and the difference was statistically significant (P<0. 05). Conclusion Both FGF-2 and tanshinoneⅡA can promote the proliferation of BMSCs and induce the differentiation of BMSCs into cardiomyocyte-like cells. The synergistic effect of the two is better than other groups.

Influence of perivascular space enlargement on the prognosis of acute anterior circulation mild stroke / 医学研究生学报

Objective Few studies are reported on the influence of perivascular space enlargement (PVSE) on the prognosis of cerebral infarction. This study was to investigate the clinical correlation of EPVS in the basal ganglia and central semiovale with the prognosis of the first acute cerebral infarction (ACI) with anterior circulation mild small-artery occlusion (SAO).Methods We treated 137 cases of the first ACI with anterior circulation mild SAO in Tangshan Gongren Hospital from August 2015 to October 2016. According to the scores on PVSE in the basal ganglia and central semiovale, we divided the patients into a mild PVSE (score: 0-1) and a severe PVSE group (score: 2-4). Based on the National Institutes of Health Stroke Scale (NHISS) and the modified Rankin Scale (MRS) scores, we classified the outcome of neurological function recovery as good (MRS≥2) and poor (MRS<2) and analyzed the risk factors for the poor prognosis of ACI by logistic regression analysis.Results There were 60 cases of severe and 77 cases of mild PVSE in the in the basal ganglia as compared with 57 cases of severe and 80 cases of mild PVSE in the in the central semiovale. Good prognosis was achieved in 97 cases while poor prognosis observed in 40. Multivariate logistic regression analysis showed that the risk factors for the poor prognosis of ACI included NHISS at the onset (OR=5.393, 95% CI: 1.858-15.654), hypertension (OR=3.729, 95% CI: 1.310-10.610), and the severity of PVSE in the basal ganglia (OR=3.137, 95% CI: 1.343-7.325).Conclusion For the first acute cerebral infarction with anterior circulation mild small-artery occlusion, the severity of PVSE in the basal ganglia is an important factor affecting the recovery of neurological function.

Clinical features and long-term prognosis of patients suffering from myocardial infarction with premature st segment elevation / 解放军医学杂志

Objective To investigate the risk factors, clinical features and prognosis of premature ST-segment elevation myocardial infarction (STEMI). Methods In this retrospective analysis, 1428 patients who were diagnosed as premature STEMI (male ≤55 years old, female ≤65) and treated with percutaneous coronary intervention (PCI), admitted to General Hospital of Shenyang Command from July 2008 to November 2012, were divided into two groups, namely the young (≤45 years old, n=381) and middle aged (>45 years old, n=1047) groups. Clinical characteristics and prognosis of the premature STEMI patients in different age groups were summarized and compared. Results The percentage of male (97.1% vs 76.7%, P<0.01), smokers (76.9% vs 65.7%, P<0.001), overweight (70.3% vs 64.9%, P=0.014), hyperlipidemia (53.3% vs 44.6%, P=0.004) were significant higher in young patients than in middle aged patients. Higher proportions of morbidities, such as hypertension (35.4% vs 43.6%, P=0.005), diabetes (14.7% vs 23.9%, P<0.001), renal insufficiency (3.4% vs 8.5%, P=0.001) and stroke (1.6% vs 5.3%, P=0.002) were found in middle aged than young patients. The incidence of multi-vessel disease was lower in young patients (47.5% vs 63.6%, P<0.001). Young patients were associated with a significantly decreased 3-year incidence of major adverse cardiovascular events compared with middle aged patients (7.1% vs 4.8%, P=0.04). Conclusion Young patients with premature STEMI is associated with less comorbidities but more controllable risk factors and worse long-term prognosis than middle aged counterparts.

Effects of acupuncture at "Tianshu" (ST 25) on electro-activity and mechanical motility of migrating motor complex during jejunal digestion period in rats with detached jejunum / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of acupuncture at "Tianshu" (ST 25) on electro-activity and mechanical motility at different phases of migrating motor complex (MMC) during jejunal digestion period in rats with detached jejunum, so as to explore the effect and mechanism of acupuncture on regulating intestinal movement.</p><p><b>METHODS</b>Sixteen adult SD rats were selected. Electrodes were implanted in the serous membrane of intestinal smooth muscl.e and high-sensitivity sensors of strain gauge were sutured on serosal surface, and then the rat was anesthetized and its jejunum was detached. Electro-acriviry and mechanical motility of jejunal smooth muscle were recorded simultaneously. Acupuncture was applied at "Tianshu" (ST 25) at MMC I , MMCII and MMC III, respectively, to observe its influence on electro-activity and mechanical motility.</p><p><b>RESULTS</b>At phase of MMC I, there was no obvious change of the fast wave before and after the acupuncture, while the frequency and amplitude of slow wave and mechanical motility were both significantly decreased compared with baseline (P < 0.01). At MMCII-Ill, the frequency and amplitude of fast wave, slow wave and motility were all significantly decreased compared with baseline (P < 0.01). Acupuncture at "Tianshu" (ST 25) had prohibited effects on electro-activity and mechanical motility of jejunal smooth muscle in rats with detached jejunum.</p><p><b>CONCLUSION</b>Acupuncture at "Tianshu" (ST 25) has obvious prohibited effects on electro-acrivity and mechanical motility at MMC I , MMC II and MMC III time phases in rats with detached jejunum. The possible mechanism is that acupuncture at "Tianshu" (ST 25) could prohibit jejunum movement through reflex path of skin-sympathetic.</p>

Effects of edaravone on the expression of GRP78, Caspase-12, and neuron apoptosis in juvenile rat hippocampus after status convulsive / 中华儿科杂志

<p><b>OBJECTIVE</b>To observe the expression of GRP78 (glucose regulated protein, GRP78), Caspase-12 and the change of neuron apoptosis in the juvenile rat hippocampus after status convulsive (SC), and to explore the effect of edaravone on them.</p><p><b>METHODS</b>One hundred and ninety-five juvenile male Sprague-Dawley (SD) rats were randomly divided into normal saline control group (NS group), status convulsive group (SC group) and edaravone treatment group (ED group). Each group was further divided into five subgroups in different executed time points after SC. The rats in status convulsive group were kindled into epilepsy by lithium-pilocarpine method. Expression of GRP78 mRNA and caspase-12 mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR) method. Expressions of GRP78 and caspase-12 protein were detected with immunohistochemical methods. The neuron apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL).</p><p><b>RESULTS</b>(1) Measured by immunohistochemistry the value of OD of GRP78 (0.1480 ± 0.0164, 0.1682 ± 0.0114, and 0.1540 ± 0.0102, respectively, 12 h - 48 h points) and caspase-12 (0.1325 ± 0.0165, 0.1794 ± 0.0213, 0.1525 ± 0.0423, and 0.1309 ± 0.0199, respectively, 12 h-72 h points) positive cells in the SC group increased, there was a significant difference compared with NS group (GRP78: 0.1214 ± 0.0147, 0.1272 ± 0.0177, and 0.1260 ± 0.0157, respectively, 12 h-72 h points. Caspase-12: 0.1050 ± 0.0121, 0.1041 ± 0.0151, 0.1058 ± 0.0222, and 0.1036 ± 0.0186, respectively, 12 h - 72 h points) (P < 0.01, or P < 0.05). By ED intervention GRP78 (0.1550 ± 0.0131, 0.1886 ± 0.0154, and 0.1721 ± 0.0151, respectively, 12 h - 48 h points) positive cells value of the OD increased as compared with SC group (P < 0.01, or P < 0.05). and caspase-12 (0.1211 ± 0.0184, 0.1545 ± 0.0205, and 0.1085 ± 0.0219, respectively, 12 h, 24 h and 72 h points) positive cells value of the A decreased as compared with SC group (P < 0.01, or P < 0.05). (2) Measured by RT-PCR, the expression of GRP78 mRNA and caspase-12 mRNA trend was similar to protein. (3) The TUNEL positive cells in hippocampus CA(1) of SC group (11.41 ± 2.37) were more than that of NS group after the SC 12 h (P < 0.01), reached its highest level at 48 h (28.78 ± 5.11), after the intervention with edaravone (8.98 ± 2.22, 13.09 ± 2.54 and 20.57 ± 4.89, respectively, 12 h-48 h points), TUNEL positive cells showed a significant drop in SC group at 12 h-48 h time points (P < 0.01, or P < 0.05), but still significantly higher than that of the NS group (6.22 ± 1.50, 6.57 ± 1.61 and 6.72 ± 1.14, respectively) (P < 0.01, or P < 0.05), at the 4 h time point (NS group 6.29 ± 1.49, SC group 6.61 ± 1.71, ED group 5.75 ± 1.41) among the three groups, no significant difference in TUNEL positive cells was found (P = 0.759).</p><p><b>CONCLUSIONS</b>The expression of GRP78 and caspase-12 increased after SC. Edaravone increased expression of GRP78 and decreased expression of caspase-12 in hippocampus rat with pilocarpine-induced seizures, reduced the number of neuronal apoptosis. These results suggest that edaravone may have protective effect against the hippocampal damage caused by status convulsive.</p>

Effects of edaravone on glial fibrillary acidic protein and interleukin-lβ expression and neuronal apoptosis in juvenile rat hippocampus after status convulsion / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of edaravone on glial fibrillary acidic protein (GFAP) and interleukin-1β (IL-lβ) expression and neuronal apoptosis in the juvenile rat hippocampus after status convulsion (SC).</p><p><b>METHODS</b>One hundred and ninety-five juvenile male Sprague-Dawley (SD) rats were randomly divided into 3 groups: normal saline control and SC with and without edaravone treatment. Each of the 3 groups was further subdivided into subgroups sacrificed at 4, 12, 24, 48 and 72 hrs after SC (n=15). The SC model was prepared using lithium-pilocarpine. The expression of GFAP and IL-lβ protein was detected with immunohistochemistry methods. The neuronal apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL). The hippocampal GFAP mRNA expression was detected by RT-PCR.</p><p><b>RESULTS</b>The value of IOD of GFAP and IL-lβ positive cells measured by immunohistochemistry in the untreated SC group increased compared with the control group. Expression of GFAP and IL-lβ protein was significantly reduced in the edaravone treated SC group compared with the untreated SC group. RT-PCR showed the expression trend of GFAP mRNA was similar to that of protein. The TUNEL positive cells in the hippocampus CA1 in the untreated SC group increased significantly 12 hrs after SC and reached a peak at 48 hrs compared with the control group. The intervention with edaravone decreased significantly TUNEL positive cells between 12-48 hrs after SC, but the number of TUNEL positive cells in the intervention group remained significantly greater than in the control group.</p><p><b>CONCLUSIONS</b>The expression of GFAP and IL-lβ in the hippocampus increases after SC in rats. Edaravone may decrease the expression of GFAP and IL-1β and reduce the number of neuronal apoptosis. These results suggest that edaravone may have protective effects against brain damage caused by SC.</p>

Protection of edaravone on neurons and its effects on the expression of interleukin-lbeta in juvenile rat hippocampus following status convulsion / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the possible protection of edaravone on neurons of the hippocampus after status convulsion (SC) and its effects on the expression of interleukin-1beta (IL-lbeta) in juvenile rats.</p><p><b>METHODS</b>One hundred and ninety-five juvenile male Sprague-Dawley rats were randomly divided into three groups: SC, edaravone pretreatment and normal saline control (control group). Each group was subdivided into five groups sacrificed at 4, 12, 24, 48 and 72 hrs after SC induction. SC model was prepared using lithium-pilocarpine. The edaravone pretreatment group received edaravone by intraperitoneal injection once daily three days before convulsion induction. Histopathologic changes in the hippocampus were viewed under a light microscope and an electron microscope. Expression of apoptosis cells was observed by TdT-mediated dUTP nick end labeling (TUNEL). Expression of IL-lbeta protein was determined by immunohistochemistry.</p><p><b>RESULTS</b>Under the electron microscrope, a small quantity of neurons showed karyopycnosis and endocytoplasmic reticulum (ER) expanded remarkably 24 hrs after SC induction; at 48 hrs the ER expanding was alleviated somewhat but mitochomdria swelling was more severe. The edaravone pretreatment group showed less severe neuronal changes compared with the SC group under the microscopes. The TUNEL positive cells in the hippocampus of the SC group were significantly more than those of the control group 12 hrs, and peaked at 48 hrs after SC induction. The edaravone pretreatment group showed decreased TUNEL positive cells in the hippocampus compared with the SC group, although the positive cells were more than those in the control group between 12 and 48 hrs after SC induction. The immunohistochemistry assay demonstrated that the expression of IL-lbeta in the hippocampus of the SC group increased significantly compared with that of the control group 12, 24, 48 and 72 hrs after SC induction. Edaravone pretreatment resulted in a significantly decreased IL-lbeta expression in the hippocampus as compared with the SC group.</p><p><b>CONCLUSIONS</b>Edaravone pretreatment may decrease the IL-1beta expression and neuronal apoptosis in the hippocampus. This suggests that edaravone may have protective effects against the hippocampal damage caused by SC.</p>

Effects of edaravone on the expression of interleukin-1beta, nuclear factor-kappaB and neuron apoptosis in juvenile rat hippocampus after status convulsion / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of edaravone on expression of interleukin-1beta (IL-1beta), nuclear factor-kappaB (NF-kappaB) and neuron apoptosis in the juvenile rat hippocampus after status convulsion (SC).</p><p><b>METHODS</b>One hundred and ninety-five juvenile male Sprague-Dawley (SD) rats were randomly divided into normal saline (NS) control group, status convulsive group and edaravone treatment group. Each group was further divided into five subgroups for different time points. The rats in status convulsive group were kindled into epilepsy by lithium-pilocarpine chemical method. Expressions of IL-1beta and NF-kappaB proteins were detected with immunohistochemistry methods. Expression of NF-kappaB mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR). The neuron apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL).</p><p><b>RESULTS</b>(1) Measured by immunohistochemistry the value of IOD of IL-1beta (30.83 +/- 3.81, 41.00 +/- 5.61, 36.32 +/- 6.78 and 28.48 +/- 4.61, respectively, 12-72 h points) and NF-kappaB (67.60 +/- 5.81, 74.61 +/- 7.94, 82.43 +/- 10.67, 70.70 +/- 5.85 and 68.22 +/- 9.67, respectively, 4-72 h points) positive cells in the SC group increased,there was significant difference compared with NS group (IL-1beta: 11.74 +/- 2.32, 12.93 +/- 2.49, 13.02 +/- 2.83 and 12.98 +/- 5.29, respectively, 12-72 h points. NF-kappaB: 48.67 +/- 16.14, 44.62 +/- 7.82, 53.16 +/- 114.45, 54.27 +/- 5.25 and 55.56 +/- 7.56, respectively, 4-72 h points) (P < 0.01, or P < 0.05). By ED intervention in IL-1beta (22.01 +/- 4.45, 28.28 +/- 4.50 and 26.00 +/- 5.34, respectively, 12-48 h points) and NF-kappaB (58.56 +/- 6.37, 59. 86 +/- 6.73, 70.00 +/- 10.09, 64.78 +/- 7.56 and 64.45 +/- 6.51, respectively, 4-72 h points) positive cells value of the IOD decreased as compared with SC group (P < 0.01, or P < 0.05). (2) Measured by RT-PCR, the expression of NF-KB mRNA and protein trend was similar. (3)The TUNEL positive cells in hippocampus, CA1 of SC group (11.41 +/- 2.37) were more than that of NS group 12 h after the SC (P < 0.01), reached its highest level at48 h (28.78 +/- 5.11), after the intervention with edaravone (8.98 +/- 2.22, 13.09 +/- 2.54 and 20. 57 +/- 4.89, respectively, 12-48 h points) ,TUNEL positive cells showed a significant drop in SC group at 12-48 h time points (P < 0.01, or P < 0.05), but still significantly higher than that of the NS group (6.22 +/- 1.50, 6.57 +/- 1.61 and 6.72 +/- 1.14, respectively) (P < 0.01, or P < 0.05), at the 4 h time point(NS group 6.29 +/- 1.49, SC group 6.61 +/- 1.71, ED group 5.75 +/- 1.41) among the three groups, no significant difference in TUNEL positive cells was found (P = 0.759).</p><p><b>CONCLUSIONS</b>Edaravone inhibited expression of IL-1beta and NF-kappaB in pilocarpine-induced seizures in rat hippocampus, reduced the number of neuronal apoptosis. These results suggest that edaravone may have protective effect against the damage caused by status convulsion.</p>

Immune response after injecting polysaccharide nucleic acid of Bacillus Calmette-Guerin under mucous membrane of bladder in rabbits / 中国肿瘤生物治疗杂志

Objective:To investigate the immune response after injecting polysaccharide nucleic acid of Bacillus Calmette-Guerin(BCG-PSN)under the mucous membrane of bladder in rabbits and to search for the most suitable dose of BCG-PSN.Methods:The rabbits were randomly divided into 5 groups:high dose(0.35 mg/ml)BCG-PSN group,mid- dle dose(0.175 mg/ml)BCG-PSN group,low dose(0.0875 mg/ml)BCG-PSN group,BCG(0.35 mg/ml)control group and BCG-PSN(0.35 mg/ml)intra-bladder perfusion control group.T lymphocyte subsets(CD4~+,CD8~+)in the peripheral blood were determined by flow cytometry before and 1 week,2 weeks,1 month and 3 months after the treatment; the levels of IL-2,TNF-?,and IFN-?of peripheral blood were determined by enzyme-linked immunosorbent assay;H-E and Masson staining was used for the pathological examination of the bladder.Results:(1)BCG-PSN injection increased the number of CD4~+ and CD8~+ T lymphocytes in a time- and dose-dependent manner,with the peak numbers appeared 2 weeks after BCG-PSN injection;the numbers restored to the normal levels 3 months after BCG-PSN injection.BCG control group had a similar changing pattern to the BCG-PSN injection group.The number of CD4~+ T cells BCG-PSN perfusion group was significantly lower than that in the high-dose BCG-PSN injection group(P

Protective effect of pueraria flavonoid on the cerebral ischemic reperfusion injury in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To research the protective effect of pueraria flavonoid on the cerebral ischemic reperfusion injury.</p><p><b>METHOD</b>Using the middle cerebral artery occlusion model (MCAO) in rats, we investigated the influence of pueraria flavonoid on the brain water content, the infarct volume, the activities of SOD, and the content of MDA.</p><p><b>RESULT</b>Pueraria Flavonoid could obviously reduce the brain water content and the infract volume in MCAO, increase the activities of SOD, and decrease the content of MDA in the cerebral ischemia- reinfusion model of rats.</p><p><b>CONCLUSION</b>Pueraria has the function of scavenging free radicals and the protective effect on ischemic brain tissue.</p>

Clinical characteristics of myasthenia gravis with dysphonia as the initial symptom / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To identify clinical features and diagnostic tests that would alert the otolaryngologist to consider myasthenia gravis (MG) in the differential diagnosis of dysphonia, we reviewed the clinical characteristics of MG whose initial symptom is dysphonia.</p><p><b>METHODS</b>31 patients who presented with dysphonia as their initial and primary complaint are reported, their symptoms and signs are observed and analyzed.</p><p><b>RESULTS</b>Patients with dysphonia as their initial symptom of MG may complain of vocal fatigue, difficulty sustaining or projecting their voices, breathy voice or intermittent hoarseness. These symptoms are characterized by fluctuating weakness and abnormal fatigability. Flexible fibroendoscopic examination revealed that patients had incomplete adduction of the vocal folds, fatigue of the tensors of the vocal fold, incomplete glottic closure, vocal cord paralysis, saliva pooling over the bilateral or unilateral pyriform sinus. Neostigmine test revealed dramatic improvement in all patients. Serum levels of anti-Ach-R antibodies were tested in 19 cases, only 5 cases were abnormality. All patients had improved after treatment</p><p><b>CONCLUSIONS</b>Voice changes can be the first sign of early MG. Based on fluctuating weakness or weak voice at the end of the day, a positive neostigmine test, significantly higher circulating antibody to acetylcholine receptor, a diagnosis of MG could definitively be made.</p>

Gene Gly82Ser polymorphism of receptor of advanced glycaton end products in patients with transient ischemia attack / 中华神经科杂志

Objective To explore the relationship between receptor of advanced glycaton end products(RAGE)gene Gly82Ser polymorphism and patients with transient ischemia attack(TIA).Methods The Gly82Ser gene at the position of RAGE gene exon 3 was identified by a polymerase chain reaction- restriction fragment length polymorphism(PCR-RFLP)method in 70 cases of TIA & Diabetes(DM), 60 of simply TIA and 66 healthy control subjects.Results The genotypes of RAGE gene Gly82Ser identified were GG, GS and SS.The frequencies of RAGE gene Gly82Ser GS heterozygous genotype of TIA & DM and control were respectively 62.9% and 43.9%, significantly higher in TIA & DM patients than in control subjects(OR 2.036, 95% CI 1.021--4.062, P=0.042), however no significant difference was found between simply TIA and control(53.3% vs 43.9%, OR 1.299,95% CI O.644--2.618, P=0.465). Significant difference of the frequency of S allele was found neither between TIA & control and control(being 34.3% and 26.5%, respectively, OR 1.446,95% CI 0.859--2.434, P=0.164), nor between simple TIA and control(28.3% vs 26.5%, OR 1.096,95%CI 0.630--1.907, P=0.746).Conclusions RAGE gene Gly82Ser GS heterozygous genotype may be associated with TIA & DM patients.RAGE gene Gly82Ser polymorphism is a risky factor for TIA & DM patients, but not for TIA patients.

Hypoxia-inducible factor-1 alpha dependent expression and significance of the related multidrug resistance genes induced by hypoxia in human hepatocarcinoma cell / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the mechanism of multidrug resistance of hepatocellular carcinoma induced by hypoxia and the potential role of hypoxia-inducible factor-1 alpha (HIF-1 alpha) and multidrug resistance related genes.</p><p><b>METHODS</b>Human hepatocarcinoma cell lines HepG2 cells were exposed to hypoxia and were transfected by plasmid HIF-1 alpha/PCDNA3, respectively. The expressions of multidrug resistance gene (mdr1), multidrug resistance protein (MRP1), and lung resistance protein (LRP) gene at the mRNA and the protein levels in the above two groups were respectively analyzed by real-time fluorescent quantitative PCR and Western-blot technique.</p><p><b>RESULTS</b>In the hypoxia group, the expressions of mdr1, MRP1 and LRP were stepped up correlating to the degree of hypoxia, especially the prominent increase in the expression of MRP1. Furthermore, they were synchronous with the changes of the expression of HIF-1 alpha. Also the increased expression of mdr1, MRP1, and LRP gene was observed in transfected HepG2 cells by plasmid HIF-1 alpha/PCDNA3.</p><p><b>CONCLUSIONS</b>Resistance of hepatocellular carcinoma to chemotherapeutics could be induced by hypoxia. HIF-1 alpha may be critical to the upregulation of the expression of the related multidrug resistance genes induced by hypoxia. HIF-1 alpha and these related multidrug resistance genes could be potential molecular targets for reversing multidrug resistance of hepatocellular carcinoma.</p>

A study on the inhibitory effect on transplanted tumor growth expressing HCV NS3 protein by HCV minigene vaccine based on invariant chain substitution in mice / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To establish a transplanted tumor producing HCV NS3 protein in mice and study the therapeutic effect of minigene vaccine based on invariant chain substitution.</p><p><b>METHODS</b>SP2/0-NS3 cells expressing HCV NS3 antigen were injected subcutaneously into BALB/ c mice. After three days of inoculation, different therapeutical reagents were injected intramuscularly into different groups of mice. The boost immunization was carried out two weeks after the first immunization. The efficiency of HCV NS3 Th1 minigene vaccine was estimated after 60 days observation.</p><p><b>RESULTS</b>For saline, pCI-neo, pHCV-NS3 and pHCV-NS3-Th1 treated groups, the induction period needed for tumor growth was 16.17+/-2.55, 14.40+/-1.82, 16.75+/-2.36, and 24.00+/-5.57 days (t =2.623, P =0.034 vs saline, t =3.713, P =0.010 vs pCI-neo and t =2.425, P =0.045 vs pHCV-NS3) respectively. The tumorigenesis rates were 100%, 100%, 57.1% (8/14, chi2 = 6.190, P = 0.013 vs saline and chi2 = 6.608, P = 0.010 vs pCI-neo) and 46.7% (7/15, chi2 = 9.707, P = 0.002 vs saline and chi2 = 10.311, P = 0.001 vs pCI-neo ) respectively. The survival rates were 0, 0, 50.0% (7/14, chi2 = 5.787, P = 0.016 vs saline and chi2 = 9.333, P = 0.002 vs pCI-neo) and 53.3% (8/15, chi2 = 6.651, P = 0.010 vs saline and chi2 = 10.311, P = 0.001 vs pCI-neo) respectively. The average tumor diameter of the pHCV-NS3-Th1 treated group was significantly smaller compared with the control groups and the pHCV-NS3 treated group (P =0.001). Moreover, the average survival time of tumor-bearing mice immunized with pHCV-NS3-Th1 was 6 days longer compared with the saline treated group, 12 days longer compared with the pCI-neo treated group (P =0.001), and 6 days compared with the pHCV-NS3 treated group.</p><p><b>CONCLUSION</b>HCV NS3 Th1 epitope vaccine might be a potential biotherapy candidate against HCV infection.</p>

Tumor necrosis factor related apoptosis-inducing ligand induces apoptosis and sensitizes resistant hepatic cancer cells to chemotherapy / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effective of tumor necrosis factor related apoptosis-inducing ligand (TRAIL) combination with chemotherapeutic agent inducing apoptosis in resistant hepatic cancer cells.</p><p><b>METHODS</b>HepG2 cells resistant to Adriamycin (HepG2/ADR) were induced stepwise. The effects of TRAIL in combination with ADM (0.1 mg/L) on promoting apoptosis in HepG2/ADR were analyzed, the proliferation was observed by MTT assay, the apoptosis of cells was also observed by flow cytometry and TUNEL method.</p><p><b>RESULTS</b>HepG2/ADR was confirmed resisting to ADM. Treated with TRAIL combination with ADM (0.1 mg/L), it showed significant inhibitory effect on the growth of HepG2/ADM, the percentage of apoptosis was increased as comparison with control at 24 h.</p><p><b>CONCLUSION</b>MDR1 might not take part in resistance to TRAIL-induced apoptosis. TRAIL dramatically augmented the sensitivity to chemotherapeutic agents in HepG2/ADR. Combined TRAIL with chemotherapeutic agents treatment could be a novel and attractive strategy to drug-resistant/ TRAIL-resistant tumor cells.</p>

Expression and clinical significance of HIF-1a protein in hepatocellular carcinoma tissues / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of HIF-1a protein in hepatocellular carcinoma (HCC) tissues.</p><p><b>METHODS</b>Immunohistochemistry (IHC), Western blotting and RT-PCR techniques were used to detect the expression of the HIF-1a gene protein in 35 HCC, 26 cirrhotic and 15 normal liver tissues. Their relationship with the pathological characteristics of the tumors were also analyzed.</p><p><b>RESULTS</b>The positive rates of HIF-1a expression in HCC tissues was 94%, which was similar to the positive rates of HIF-1a expression in liver cirrhosis tissues of 92%, but was higher than that in normal hepatic tissues of 7%, but the residual proliferatic hepatic trabeculae among the necrotic liver cells and the fibrotic tissues expressed HIF-1a strongly in comparison with the cirrhotic liver tissues. The expression intensity of HIF-1a protein of the cirrhotic liver tissues was stronger than that in HCC; the results by Western blotting and RT-PCR were in accordance to that by IHC. In addition, the expression intensity in HCC had a negative correlation in differentiation degree and a positive correlation to intrahepatic and extrahepatic metastases but no correlation was found between HIF-1a expression and the existence of portal vein tumor emboli, prognosis and the status of HBsAg.</p><p><b>CONCLUSION</b>HIF-1 protein was expressed in HCC and cirrhotic liver tissues, and was only affected by the factor of hypoxia. The expression of HIF-1a protein is associated with the differentiation of the tumor and its intrahepatic and extrahepatic metastases but was not related to the existence of portal vein tumor emboli, prognosis and the status of HBsAg. This phenomenon may provide a new idea for the treatment of liver cancer.</p>

Antitumor effects of soluble TRAIL in human hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate therapeutic potential of soluble TRAIL (sTRAIL) in hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Expression of TRAILR was determined by in situ hybridization in 60 samples of resected hepatocellular carcinoma, 20 samples of normal liver tissue near the margin of benign tumor and 2 HCC cell lines of HepG2 and SMMC-7721. The clinical data of the patients were analyzed as well as cellular effects of sTRAIL in promoting apoptosis on HCC cell lines HepG2 and SMMC-7721 (p53 gene mutated) after exposure to different concentrations of recombinant protein.</p><p><b>RESULTS</b>High death receptor (DR) expression and low DcR expression in HCC tissue differed from low DR expression and high DcR expression in the normal hepatic tissue with statistical significance. DR5, DR4, and DcR2 but not DcR1 were expressed in both cell lines. The expression of DR was closely correlated with HCC differentiation, with the weak expression in poor differentiation. The positive rate of DR expression in 32 cases of grade III-IV was significantly lower than that in 28 cases of grade I-II (P < 0.05). Cell apoptosis rates were 10%, 70% and 50% of HCC cells, Jurkat cells and human cholangiocarcinoma cell line QBC939 24 h after recombinant of TRAIL alone.</p><p><b>CONCLUSION</b>TRAILR expression is prevalent in HCC, with different receptor types existing. HCC is resistant to TRAIL-mediated apoptosis. The treatment of TRAIL alone only has a limited effect on inducing apoptosis on HCC cell lines of HepG2 and SMMC-7721.</p>

Hepatitis B x protein activated vascular endothelial growth factor expression through hypoxia inducible factor-1 pathway / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate whether hepatitis B x protein (HBx) stimulates vascular endothelial growth factor (VEGF) through hypoxia inducible factor-1 (HIF-1 alpha) pathway.</p><p><b>METHODS</b>Two plasmids including pIRES-EGFP-HBx and pTK-Hyg were co-transfected to a hepatocellular carcinoma cell line SMMC-7721. With fluorescence-positive and fluorescence-negative hygromycin-resistant colonies selected, expressions of VEGF and HIF-1 alpha in protein or/and mRNA level were detected.</p><p><b>RESULTS</b>Fluorescence-positive cells were stably integrated with HBx, in which expression of HIF-1 alpha and VEGF were upregulated. Fluorescence-negative cells did not express HBx, VEGF or HIF-1 alpha.</p><p><b>CONCLUSION</b>HBx can activate VEGF through HIF-1 alpha pathway.</p>