OBJECTIVE To investigate the effect and potential mechanism of sodium ferulate （SF） on corneal endothelial dysfunction and corneal endothelial cell （CEC） injury. METHODS The male New Zealand rabbits were divided into control group， benzalkonium chloride （BAK） group and BAK+SF group， with 6 rabbits in each group. Except for control group， the other groups were given BAK into the anterior chamber to induce bullous keratopathy model， and BAK+SF group then given SF solution 200 mg/kg intraperitoneally the next day after surgery， twice a day， for consecutive 14 d. The transparency of corneal and edema of corneal stroma in each group of rabbits （before and on the 1st， 7th， and 14th day after surgery） were observed， and the corneal thickness （14th day after surgery） and intraocular pressure （1st to 14th day after surgery） were measured. On the 14th day after operation， the corneal endothelial structure was evaluated and the expressions of functionally related proteins [phalloidin， zonula occludens-1 （ZO-1）， Na+/K+-ATPase， Ki67] were detected. On the 14th day after surgery， the corneal tissue was collected in BAK group， the primary rabbit CECs were isolated and cultured， and they were divided into blank group and SF groups with different mass concentrations. The cell viabilities after being cultured for different time， and the protein expressions of Ras homologous gene family A （RhoA）， bone morphogenetic protein receptor 1A （BMPR1A） and BMRP2 were determined in each group. RESULTS Compared with BAK group， the transparency of corneal and edema of corneal stroma were gradually improved， and the corneal thickness was significantly decreased in BAK+SF group （P＜0.05）. The rabbit CECs in BAK+SF group were only damaged to zone B and showed a normal hexagonal endothelial cells structure. The protein expressions of phalloidin， ZO-1， Na+/K+-ATPase and Ki67 in BAK+SF group were significantly increased （P＜0.05）. When SF concentration was lower than and equal to 200 mg/L， it could promote the proliferation of rabbit CEC， in concentration manner （P＜0.05） and time-dependent trend. SF at concentrations of 50， 100， and 200 mg/L could up-regulate the protein expressions of RhoA， BMPR1A and BMPR2 in concentration-dependent manner （P＜0.05）. CONCLUSIONS SF can improve the transparency of corneal and edema of corneal stroma in bullous keratopathy model rabbits， reduce corneal thickness， maintain the integrity of corneal endothelium structure， and promote the recovery of corneal endothelial function； this compound can promote the proliferation of CEC， the mechanism of which may be related to the activation of RhoA-ROCK-BMP pathway.

ObjectiveTo investigate the mechanism of action of PNPLA3 I148M mutation in the development and progression of non-alcoholic fatty liver fibrosis. MethodsThe lentiviral vectors carrying the mutant or wild-type PNPLA3 I148M gene were constructed and transfected into rat hepatic stellate (HSC-T6) cells. Quantitative real-time PCR was applied to measure the mRNA expression of transforming growth factor β1 (TGF β1). The t-test was applied for statistical analysis. ResultsThe lentiviral vectors carrying the mutant or wild-type PNPLA3 I148M gene were successfully constructed and transfected into HSC-T6 cells, and a HSC-T6 cell line with stable expression of the mutant or wild-type PNPLA3 gene was established. Compared with the cell line carrying the wild-type gene, the cell line carrying the mutant gene showed significantly higher mRNA expression of TGF β1 (1.25±0.15 vs 0.48±0.07; t=11.826, P＜0001). ConclusionPNPLA3 I148M mutation can increase the expression of TGF β1 in HSC-T6 cells, which provides a new cell model and new research ideas for investigating the role of PNPLA3 I148M mutation in non-alcoholic fatty liver fibrosis.

Objective To investigate the influence of leptin receptor (LEPR) gene K109R polymorphism on the risk of nonalcoholic fatty liver disease (NAFLD) and its interaction with PNPLA3 I148M polymorphism in the Han Chinese population in Qingdao,China.Methods Blood samples were collected from 296 NAFLD patients and 321 healthy controls,and the genotypes of these patients were determined by PCR and genotyping.Related statistical analyses were performed to compare genotypes,alleles,and clinical data between the two groups.Generalized multifactor dimensionality reduction (GMDR) was used to investigate the interaction between LEPR K109R and PNPLA3 I148M genes.Results The distribution of LEPR K109R genotypes and alleles showed no significant differences between the NAFLD group and the control group (P ＞ 0.05).PNPLA3 I148M gene polymorphisms were closely associated with the risk of NAFLD,and the risk of NAFLD in G mutant gene carriers was 2.07 times that in patients who did not carry this gene (OR =2.07,95％ CI 1.423-3.013,P ＜ 0.001).The joint action of LEPR K 109R and PNPLA3 I 148M significantly increased the risk of NAFL (OR =3.393,95％ CI 1.856-6.201,P ＜ 0.001).Conclusion In the Han Chinese population in Qingdao,LEPR K109R gene polymorphism is not associated with the risk of NAFLD,but its interaction with PNPLA3 I148M polymorphism can significantly increase the risk of NAFLD.

OBJECTIVETo investigate the relationship between SREBP-1c and the risk of liver disease associated with the triacylglyceride lipase PNPLA3 I148M variant using a human hepatoma cell line model transfected with recombinant lentiviruses.

METHODSHuh7 cells were transfected with control lentivirus or lentivirus containing the PNPLA3 I148M variant (variant). The two cell groups were compared to assess differences in triglyceride content (using oil red O staining), levels of triglyceride and cholesterol (using automated biochemical analyzer), expression of SREBP-lc mRNA (using fluorescence quantitative PCR), and expression of SREBP-1c protein (using western blot.

RESULTSCells expressing the PNPLA3 I148M variant showed higher triglyceride content (0.54+/-0.03 mmol/L vs. control cells: 0.23+/-0.02 mmol/L; t=22.58, P<0.001), cholesterol level (0.28+/-0.03 mmol/L vs. control cells: 0.13+/-0.02 mmol/L; t =11.83, P<0.001), SREBP-1cmRNA expression (13.59+/-0.60 vs. 11.81+/-0.82; [The abstract and text in the paper say variant increases, but the data shown says the higher value is in the control cells. Please correct to properly express the data.] P=0.001), and SREBP-1c protein expression. The level of SREBP-1c was positively correlated with serum triglyceride in the cells expressing the PNPLA3 I148M variant (r=0.912, P<0.01).

CONCLUSIONThe risk of liver disease associated with the PNPLA3 I148M variant, which increases lipogenesis, may involve SREBP-1c and a pathway that increases triglycerides.

Objective To investigate the association between the Adiponectin rs266729 and rs2241766 gene polymorphisms and nonalcoholic fatty liver disease in the Han Chinese population residing in Qingdao. Methods Adiponectin rs266729 and rs2241766 gene polymorphisms were genotyped in patients with NAFLD (n = 336) and healthy controls (n = 280) using polymerase chain reaction (PCR). Serum lipid profiles and adiponectin levels were determined using biochemical methods. Statistical analyses were performed using Pearson Chi square test, logistic regression analysis, t test, linear regression analysis. Results We found a significant association between the Adiponectin rs266729 genotype frequencies and allele frequencies between NAFLD pa-tients and controls (χ2= 9.929, P = 0.007; χ2= 9.809, P = 0.002). After adjustment of confounding factor, the rs266729 G allele was associated with an increased risk of NAFLD compared to the C allele (OR = 1.410, 95%CI: 1.082-1.831, P = 0.008) No significant differences were found in the rs2241766 genotype frequencies and allele frequencies between NAFLD population and the controls (OR = 1.410, 95%CI: 1.082-1.831, P = 0.008). Conclusion The Han Chinese in Qingdao carrying the rs266729 G allele are at increased risk of NAFLD.

Objective To explore the relationship between traditional Chinese medicine (TCM) syndromes of state of evil domination and clinical and laboratory indicators of IgA nephropathy. Methods A prospective study was used to collect data on clinical and laboratory examination of IgA nephropathy in multi clinical centers. Patients’ TCM syndrome types were determined according to the national diagnostic criteria at the same time. Totally 266 patients with IgA nephropathy were included in the study to establish a database for analyzing the relationship between TCM syndrome of state of evil domination and clinical and laboratory indicators. Results In the relationship between syndromes of state of evil domination distribution and clinical subtypes, patients of wind-heat syndrome had more macroscopic hematuria;patients of phlegm-damp syndrome had more nephrotic syndrome;patients of damp-heat syndrome had more chronic nephritis type Ⅰ;patients of blood stasis syndrome had more chronic nephritis type Ⅱ. In the relationship between syndrome of state of evil domination distribution and clinical manifestation, the incidence of hypertension was higher in patients of blood stasis syndrome than in other three types. It was more serious for hematuresis in the patients of wind-heat syndrome. For patients of phlegm-damp syndrome, the incidence of heave proteinuria was highest. In the relationship between syndrome of state of evil domination distribution and laboratory examination, 24-hour urinary protein quantification was higher than in patients of wind-heat and damp-heat syndrome, but the level of blood albumin was lowest. For patients of blood stasis syndrome, serum creatinine level was significantly higher than in other three types;the level of eGFR was just the opposite. The levels of blood cholesterol and triglyceride in patients of phlegm-damp syndrome were higher than in other three types. The activated partial thromboplastin time (APTT) levels in patients of blood stasis and phlegm-damp syndrome were lower than in other patients, but the FIB level was the exact opposite of APTT. In the relationship between syndrome of state of evil domination distribution and the stages of chronic kidney disease (CKD), patients of wind-heat syndrome were more in the first stage of CKD;patients of blood stasis syndrome were more in the third stage of CKD. Conclusion There is relative correlation between TCM syndromes of state of evil domination and clinical and laboratory indicators in IgA nephropathy, which would provide some reference to narrow the gap in the information of the four methods of TCM with clinical and laboratory indicators to enhance accurate diagnosis of TCM syndrome.

OBJECTIVETo study chemical constituents in nutshell of Trapa acornis and in vitro inhibitory activity against alpha-glucosidase.

METHODEtOAC and n-butanol extractive fractions were separated by chromatography and their structures were identified by multiple spectroscopic techniques.

RESULTNine compounds were separated, they were 4,23,24-trimethylcholest-22-en-3-ol (1), stigmasterol (2), alpha-amyrin (3), (+)-nyasol (4), oleanolic acid (5), ursolic acid (6), hederagenin (7), 3,23-dihydroxy-12-ursen-28-oic acid (8) and beta-daucosterol (9). Total extracts from T. acornis nutshells, petroleum ether fractions, acetic ether fractions and normal butanol fractions showed inhibitory activity against alpha-glucosidase. Compound 5 (IC50 2.88 mg x L(-1)) and 6 (IC50 4.42 mg L(-1)) showed stronger inhibitory activity against alpha-glucosidase.

CONCLUSIONAll compounds except compound 2 were separated from the genus for the first time, and compound 1-9 were separated from this plant for the first time.

ObjectiveTo observe the effect of cerebellar fastigial nucleus treated by electrical stimulation on patients with acute cerebral infarction.Methods140 cerebral infarction patients with hemiplegia were randomly divided into the experimental group and control group with 70 cases in each group.The experimental group was treated with routine drugs and electrical stimulating on cerebellar fastigial nucleus;the control group was only treated with routine drugs.The changes of motor function of limbs and activities of daily living of two groups were evaluated after treatment.ResultsAfter 21 days treatment,the score of Fugl-Meyer assessment of the experimental group was 39.2±(7.7) and that of the control group was 26.3±8.1.There was a significant difference between two groups(t=4.22,P<0.01).The score of Barther index of the experimental group was 43.8±8.4 and that of the control group was 29.7±7.5.There was also a significant difference between two groups(t=4.97,P<0.01).ConclusionThe electrical stimulation on cerebellar fastigial nucleus is a new effective way to treat acute cerebral infarction.

Aim To study the effect of Lacidophilus exopolysaccharides(LAEPS) on immunity. Methods Effects of LAEPS on immunity were investigated by delayed type hypersensitivity reaction, haemolytic plaque assay and macrophage function assay in mice. Results LAEPS ip 7～8 day promoted delayed type hypersensitivity reaction, increased haemolytic plaque and enhanced macrophage function in a dose-dependent manner.Conclusion LAEPS is able to enhances immunity.

Objective: To search for a best method for management mandible fractures by evaluating the effects of different treatments.Methods: We retrospectively analyzed 148 cases of mandible fractures treated in our department from January 1996 to June 2007.Results: Among the total number,134 cases were restored to normal occlusion,while 6 cases experienced local occlusive disfunction and 8 malocclusion.The effect of treatment was correlated with the types of fracture and methods of diaplasis.Conclusion: Mandible fractures should be treated with a new concept of combined and sequential multidisciplinary methods.Sound diaplasis followed by reliable fixation can produce a satisfying curative effect.At present,intermaxillary elastic traction with internal titanium plate fixation is the most effective method for the management of mandible fractures.