Objective To investigate the features of familiar facial palsy,ophthalmoplegia and dysphagia characterized by autosomal dominant inheritance in a family and to discuss the classification and pathogenesis of the disease.Methods Clinical,electrophysiological,pathological examinations were performed and blood samples were obtained from 5 patients and 26 family members.PCR protocol was used to identify a certain gene. Results In the 5 patients receiving physical examination,all had ptosis,external ophthalmoplegia,facial paralysis,dyphagia,hoarseness,decreased pharyngeal reflex;4 had amyotrophy of muscle of tongue,temporal nuscle,masseter and muscles of distal lower limbs;3 had proximal limb asthenia and distal limbs amyotrophy.Compared to those of oculopharyngeal muscular dystrophy(OPMD)with similar symptoms and signs,both electrophysiological manifestation and pathological findings of the family members supported the diagnosis of muscular dystrophy,but the(GCG)6(GCA)3GCG in the first exon of PABPN1 mutated neither in normal family members nor in patients.Conclusions This family presents clinical manifestations somewhat resembling to those of OPMD and distinctive to other disorders,but has a totally different genetic background from OPMD.It may be a new subtype of muscular dystrophy.

OBJECTIVETo investigate the growth inhibition and apoptosis of gastric cancer cell MKN45 induced by oridonin and its mechanism.

METHODSThe MTT method was used to investigate the inhibitory effect of oridonin on MKN45 cells. The AO/EB and Hoechst 33258 staining were used to observe the cell morphologic changes of apoptosis induced by oridonin. Prophase apoptotic ratio and cell cycle change were evaluated by GuavaEasycyte PCA-96 system. The expressions of Bcl-2, Bax and caspase 3 proteins were determined by Western blot.

RESULTSOridonin significantly inhibited the proliferation of MKN45 cells in dose- and time-dependent manner. Typical apoptotic features of the cells treated with oridonin were found by AO/EB and Hoechest33258 staining. When MKN45 cells were treated with different doses of oridonin for 12 h, the prophase apoptotic ratio was stepped up from 3.3% (untreated group) to 8.7%-17.9%; after 24 h, from 4.8% (untreated group) to 13.9%-29.3%. There was significant difference between treated and untreated groups (P <0.01). After treatment with oridonin for 24 h, MKN45 cells were arrested at G(2)/M phase. Western blot analysis showed up-regulated expression of Bax and caspase-3, and no significant change of Bcl-2, but Bcl-2/Bax ratio decreased significantly.

CONCLUSIONSOridonin significantly inhibits the proliferation of MKN45 cell. Apoptosis of MKN45 induced by oridonin may be associated with the up-regulated expression of Bax and the change of Bcl-2/Bax ratio, thus to activate the caspase pathway.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Kaurane ; pharmacology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Stomach Neoplasms ; metabolism ; bcl-2-Associated X Protein ; metabolism

BACKGROUND: Yaotu Granules have been proved to protect human nucleus pulposus cells and delay their degeneration. Notably, Yaotu Granules for lumbar disc herniation has achieved good clinical results.OBJECTIVE: To investigate the effect of the herbal compound formula Yaotu Granules on the Fas/FasL expression in a rabbit model of lumbar disc degeneration, and further elucidate the underling mechanism of preventing and treating lumbar disc degeneration.METHODS: Twenty New Zealand white rabbits were enrolled and the models of lumbar disc degeneration were established by minimally invasive puncture and rotation cutting, followed by randomized into normal saline, low-, middle-,and high-dose groups (n=5 per group). 10 mL of normal saline, 10, 20, and 40 mL of water decoction of Yaotu Granules were administered intragastrically into the normal saline, low-, middle-, and high-dose drug groups for 21 days, twice daily, respectively. Subsequently, the expression level of Fas/FasL in the rabbit nucleus pulposus cells in each group was detected.RESULTS AND CONCLUSION: The signal intensity of the rabbit lumbar disc on MRI was decreased, and ruptured annulus and posterior herniated disc were visible at 12 weeks after modeling. Masson staining showed that the nucleus pulposus cells arranged in disorder, and even ruptured. Additionally, safranin O staining found that the number of nucleus pulposus cells was decreased obviously. The order of the relative expression levels of Fas and FasL mRNA in the nucleus pulposus cells was as follows: normal saline group > low-dose drug group > middle-dose drug group > high-dose drug group (P < 0.05). These results suggest that Yaotu Granules delay the rabbit lumbar disc degeneration by downregulating the expression level of Fas/FasL.

Objective To explore how to apply the teaching mode combined flipped class with PBL teaching method to improve college students' ability of English reading and writing and the effectiveness of classroom. Methods In the class of college English reading and writing, students at grade 2015 were randomly divided into two groups. The experimental group (43 students) was adopted the class teaching with " flipped class and PBL teaching method"and the control group (43 students) was adopted traditional class teaching mode taught by the same teacher. The teaching effects of two groups were studied in a comparative way. The students' final grade, related data in the learning process, students'view of the two teaching modes are studied by analysis of basic data and questionnaire. Results At the end of the semester, the results of the experimental group are better than those of the control group. In the survey of the two class teaching modes, the students in the experimental group spent more time in the learning process than that of the control group.Students think that the teaching mode combined flipped class and PBL teaching method not only improves their autonomic learning ability, classroom participation, the English reading and writing ability, but also helps them perform better in the average scores in the final exam compared to the traditional classroom. Students are more satisfied with the new classroom teaching mode than with traditional one. Conclusion The teaching mode combined flipped class with PBL teaching method can promote the students'autonomous learning, contribute to absorb and internalize knowledge, improve the students'learning interest and learning efficiency, and improve teachers teaching ability.

Objective To compare and analyze the epidemiological features of adult patients with distal femoral fracture between the east and west areas in China from 2010 to 2011.Methods The data of adult patients with distal femoral fracture treated from January 2010 through December 2011 in 63 hospitals were collected through the PACS system and case reports checking system.The data from 35 hospitals in the east area were classified as group A and those from 28 hospitals in the west area as group B.The analytic items included gender,age and AO classification.Results A total of 2,523 adult distal femoral fractures were collected,involving i,544 males and 979 females,with a male to female ratio of 1.58:1.The adult femoral fractures predominated in an age range from 41 to 50 years (18.94％) and their high-risk type was 33-A (50.18％).In group A of 1,650 cases,there were 1,027 males and 623 females,with a male to female ratio of 1.65:1 and a median age of 48 years;in group B of 873 cases,there were 517 males and 356 females,with a male to female ratio of 1.45:1 and a median age of 45 years.There were no significant differences in the age distribution and gender proportion between groups A and B (P ＞ 0.05).The age distribution showed that the peak ranges were from 31 to 60 years in both groups.The proportion of type 33-A was the most and that of type 33-B the least in both groups.The proportion of type 33-A in group B (53.49％) was significantly higher than in group A (48.42％) while that of type 33-B in group B (16.84％) was significantly lower than in group A (21.39％) (P ＜ O.05).Conclusions Adult distal femoral fractures were common in middle-aged males and their high-risk type was 33-A.Their peak age was from 31 to 60 years in both east and west areas in China.They were more common in men.The predominant fracture type was 33-A in both areas.The east area witnessed a significant higher proportion of type 33-B and a significant lower proportion of type 33-A than the west area.

OBJECTIVETo investigate the role of tissue factor (TF) expression in the invasive and metastatic ability of colorectal carcinoma and explore the influence of TF on the invasive ability of HT-29 cells.

METHODSTF expression of specimens from 85 colorectal carcinomas and 6 colorectal adenomas was observed by immunohistochemistry. The role of TF expression in prognosis and tumor invasion and metastasis was analyzed. The plasmids pcDNA3.1/Zeo bearing either sense or antisense-TFcDNA were transfected into HT-29 cells by the way of Lipofectamine 2000. TF proteins in transfected and untransfected HT-29cells were detected by Western blot. In vitro Matrigel invasion assays were performed to show the invasive ability of those cells.

RESULTSTF expression was positive in 40 (47.1%) of 85 colorectal carcinoma specimens, but negative in normal mucosa and adenoma specimens. TF expression showed significant correlation with tumor invasive depth (r = 0.895, P < 0.01). TF expression showed significant correlation with synchronous and metachronous hepatic metastasis (r = 0.974, P < 0.01 and r = 0.963, P < 0.01 respectively). TF expression was a significant risk factor for hepatic metastasis (P < 0.01) and prognosis (P < 0.01). TF expression in HT-29 cells with sense/antisense-TFcDNA transfection was more/less than that of the cells without transfection. The invasive ability of HT-29 cells with sense-TFcDNA transfection was increased in vitro compared with the untransfected cells, but HT-29 cells with antisense-TFcDNA transfection got the contrary change.

CONCLUSIONSTF may take part in the invasive and metastatic process of primary colorectal carcinoma, and TF expression may be an indicator of hepatic metastasis and prognosis for colorectal carcinoma patients. TF expression may increase the invasive ability of HT-29 cell in vitro.

Blotting, Western ; Cell Movement ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; HT29 Cells ; Humans ; Immunohistochemistry ; Logistic Models ; Multivariate Analysis ; Thromboplastin ; analysis ; genetics

OBJECTIVETo study the clinical efficacy of three kinds of hybrid bioartificial liver support systems (HBLSS) in treating chronic severe hepatitis.

METHODSA bioartificial liver support system (BAL), comprising porcine hepatocytes and fiber tube style bioreactor, was constructed. Then three kinds of HBLSS were constructed: Molecular absorbent recirculating system (MARS) plus BAL; slow plasma exchange (SPE) plus continuous hemodiafiltration (CHDF) and BAL; and SPE plus hemoperfusion (HP) and BAL. One hundred-twenty patients in middle or late stages of chronic severe hepatitis were enrolled in this study. They were randomly divided into 6 groups: H1 group was treated with BAL+MARS, H2 with BAL+SPE+CHDF and H3 with BAL+SPE+HP (as treatment groups); C1 group was treated with MARS, C2 with SPE+CHDF and C3 with SPE+HP (as control groups). The changes in the clinical symptoms, in the hepatic encephalopathy stages, and in the serum total bilirubin (TBIL), the serum albumin (ALB), the prothrombin activities (PTA), endotoxin, ammonia, creatinine and a-fetal protein (AFP) were all observed before the treatment, right after it and 72 hours later. The improving and curing rates and the rates of side effect occurrences in each group were observed.

RESULTSIn all 6 groups, the patients' clinical symptoms ameliorated; their TBIL, endotoxin and ammonia levels decreased (P<0.05), and their PTA and AFP levels lowered significantly (P<0.05). But in the H1, H2 and H3 groups they were more distinctive than in the control groups. In H1 and H2 groups creatinine and ammonia levels were decreased more significantly than in the H3 group (P<0.05). The improving and curing rates of each group were 65 % (13/20), 60% (12/20), 45% (9/20), 45% (9/20), 40% (8/20) and 20% (4/20) respectively. No serious side effects were observed during the treatment.

CONCLUSIONIn treating middle and late stage chronic severe hepatitis, the measures used in H1, H2 and H3 are better than those in C1, C2 and C3. Furthermore, H1 and H2 treatments can ameliorate hepatic and renal functions, prevent the development of multiple organ dysfunction syndrome, and are better than those used in H3.

Adult ; Aged ; Animals ; Bioreactors ; Critical Illness ; Female ; Hemodiafiltration ; Hepatic Encephalopathy ; blood ; therapy ; Hepatitis, Viral, Human ; therapy ; Humans ; Liver ; cytology ; Liver Failure, Acute ; therapy ; Liver, Artificial ; Male ; Middle Aged ; Plasma Exchange ; Swine

OBJECTIVETo investigate the effects of the self-developed anti-heparanase polypeptide antibodies on growth and invasion of human hepatocellular carcinoma HCCLM6 cells.

METHODSUsing MTT, flow cytometry, plate clone formation, transwell invasion and heparan degrading enzyme assay, the growth and invasion changes of human hepatocellular carcinoma HCCLM6 cells by co-culture with each of three self-developed rabbit anti-heparanase polyclonal antibodies were detected.

RESULTSCompared with normal rabbit IgG, in the presence of each anti-heparanase polypeptide antibody, the growth, cell cycle and clone formation remained unchanged, and under the P1 or P2 anti-heparanase polypeptide antibody (with final concentration 100 microg/ml), the cell invasiveness was inhibited by 52.5% and 36.6%, respectively, and the heparanase activity was inhibited by 42.9% and 39.1%, respectively.

CONCLUSIONThe P1 and P2 anti-heparanase polypeptide antibodies can effectively inhibit the invasion ability and heparanase activity of liver cancer HCCLM6 cells. However, All the three antibodies have no effects on its growth, cell cycle and clone formation.

Antibodies ; pharmacology ; Carcinoma, Hepatocellular ; enzymology ; pathology ; Cell Adhesion ; Cell Differentiation ; Cell Line, Tumor ; Cell Movement ; Coculture Techniques ; Enzyme Activation ; Glucuronidase ; immunology ; metabolism ; Humans ; Liver Neoplasms ; enzymology ; pathology ; Neoplasm Invasiveness

Donation after brain death followed by circulatory death (DBCD) is a unique practice in China. The aim of this study was to define the pathologic characteristics of DBCD liver allografts in a porcine model. Fifteen male pigs (25-30 kg) were allocated randomly into donation after brain death (DBD), donation after circulatory death (DCD) and DBCD groups. Brain death was induced by augmenting intracranial pressure. Circulatory death was induced by withdrawal of life support in DBCD group and by venous injection of 40 mL 10% potassium chloride in DCD group. The donor livers were perfused in situ and kept in cold storage for 4 h. Liver tissue and common bile duct samples were collected for hematoxylin and eosin staining, TUNEL testing and electron microscopic examination. Spot necrosis was found in hepatic parenchyma of DBD and DBCD groups, while a large area of necrosis was shown in DCD group. The apoptosis rate of hepatocytes in DBD [(0.56±0.30)%] and DBCD [(0.50 ± 0.11)%] groups was much lower than that in DCD group [(3.78±0.33)%] (P<0.05). And there was no significant difference between DBD group and DBCD group (P>0.05)). The structures of bile duct were intact in both DBD and DBCD groups, while the biliary epithelium was totally damaged in DCD group. Under electron microscope, the DBD hepatocytes were characterized by intact cell membrane, well-organized endoplasmic reticulum, mild mitochondria edema and abundant glycogens. Broken cell membrane, mild inflammatory cell infiltration and sinusoidal epithelium edema, as well as reduced glycogen volume, were found in the DBCD hepatocytes. The DCD hepatocytes had more profound cell organelle injury and much less glycogen storage. In conclusion, the preservation injury of DBCD liver allografts is much less severe than that of un-controlled DCD, but more severe than that of DBD liver allografts under electron microscope, which might reflect post-transplant liver function to some extent.

BACKGROUND: Methylene blue is used as a developer to identify intervertebral disc degeneration in the transforaminal endoscopic surgery. However, many scholars have indicated that methylene blue can accelerate the degeneration process, whilst foreign researches have reported that it may play therapeutic effect on degenerative intervertebral discs under acidic conditions due to its acidophily. Therefore, whether methylene blue holds toxic effect on the disc remains controversial. OBJECTIVE: To determine whether methylene blue exerts toxic effect on nucleus pulposus cells by cell counting-kit 8 (CCK-8) assay. METHODS: The discarded nucleus pulposus from two patients with intervertebral disc herniation were selected. After digestion, nucleus pulposus cells were extracted and cultured until proliferated to 80% of the medium. Then, the cells were digested to make cell suspensions, divided into six groups and inoculated into the 96-well plates: blank control (only the medium, CCK-8 solution), control group (only medium, cells and CCK-8 solution), and the other groups were cultured with 1%, 2%, 3%, and 4% methylene blue, respectively. The absorbance values were measured by CCK-8 assay at 24, 48, 72 and 96 hours after incubation to calculate the cell viability, and the color change was observed. RESULTS AND CONCLUSION: The color in the control group was the deepest, and the color became lighter with the concentration of methylene blue increasing. The cell viability was the highest in the control group, and it was decreased with the concentration of methylene blue increasing. Thus, methylene blue may exert toxic effect on human nucleus pulposus cells.