OBJECTIVETo investigate the effects of hydrodynamics-mediated RNAi for Mfn2 gene expression in liver and the levels of blood sugar and fat in mice.

METHODSFifty-six male BALB/c mice were randomly divided into normal control group (NC, n = 8), negative control group (HK, n = 24) and transfection group (Mfn2, n = 24) according to random digits table. 1.5 ml plasmid (negative control or Mfn2 shRNA, 75mug for each mouse) diluted into phosphate buffered solution (PBS) was injected into the HK and Mfn2 groups mice via hydrodynamic intravascular injection. Mfn2 mRNA and protein expression in hepatic tissue was detected by RT-PCR and Western-blot 24 hours, 72 hours and 120 hours respectively after injection. At the same time, the levels of fasted blood sugar (FBS) and triglyceride (TG) were measured.

RESULTSCompared with HK mice, the expressions of Mfn2 mRNA (1.00+/-0.03 vs 1.14+/-0.07, t = 4.027, P = 0.007; 1.01+/-0.053 vs 1.18+/-0.07, t = 4.234, P = 0.006) and protein (7.81+/-0.80 vs 8.01+/-0.08, t = 2.941, P = 0.042; 8.05+/-0.15 vs 8.56+/-0.014, t = 4.883, P = 0.039) decreased markedly in Mfn2 mice in 72 and 120 hours after injection. In the fasting state, in 24 hours after injection, FBS in Mfn2 group was significantly lower than that in HK group [(2.65+/-0.70 vs 5.28+/-0.82) mmol/L, t = 6.879, P value less than 0.01] and TG was also significantly higher than that in HK group [(1.96+/-0.32 vs 1.12+/-0.16) mmol/L, t = -6.711, P value less than 0.01]. No statistical differences found between the NC and HK groups for FBS and TG (F = 1.412, P = 0.26; F = 2.711, P = 0.14). The plasma glucose level in Mfn2 mice was significantly higher than that in HK mice [(7.23+/-0.82 vs 5.18+/-0.69) mmol/L, t = 2.050, P value less than 0.01; (7.00+/-0.67 vs 6.05+/-0.76) mmol/L, t = 3.57, P = 0.023] in 72 and 120 hours after injection. However, no differences found between the two groups for blood TG [(1.53+/-0.27 vs 1.37+/-0.18) mmol/L, t = 0.160, P = 0.23; (1.84+/-0.30 vs 1.52+/-0.37) mmol/L, t = 0.330, P = 0.503].

CONCLUSIONThe data indicate that hydrodynamics- mediated RNAi for Mfn2 gene can effectively inhibit the expression of target gene in mice liver in 72 and 120 hours after shRNA administration, and the inhibition of hepatic Mfn2 can induce glycometabolic and fat metabolic disorder.

Animals ; Blood Glucose ; metabolism ; GTP Phosphohydrolases ; genetics ; metabolism ; Gene Expression ; Hydrodynamics ; Lipids ; blood ; Liver ; chemistry ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; RNA Interference ; RNA, Messenger ; genetics

Objective To investigate the expression level of CD226 mRNA in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE) and explore the relation between the gene expression and disease activity, and the relation between the gene expression and Gly307Ser polymorphism of CD226 was also examined. Methods CD226 gene was measured with real-time polymerase chain reaction (qRT- PCR) in PBMCs. The expression levels of CD226 gene in PBMCs were compared between 90 SLE patients and 30 healthy individuals. One-way ANOVA and pearson correlation were used for statistical analysis. Results The expression level of CD226 in the PBMCs of SLE patients (6.8±1.1) was significantly decreased compared to healthy individuals (26.5±6.7) (P＜0.01), while there was no association between mRNA level and genotype (P＞0.05). No correlation between ESR, CRP, ANA, SLEDAI scores, C3 and the expression level of CD226 gene was discovered. Conclusion In Hubei Chinese Han population, CD226-Gly307Ser locus is associated with the development of SLE, while T allele does not impact the expression of CD226 gene, thus the role of CD226 gene in autoimmune diseases should be explored in the future.

In this paper, three spoilage organisms were separated from five transmutative soy milks, and all the three spoilage bacteria could survive condition of both 1?105Pa,30min and 300mg/kg Nisin. Morpha character, physiological and biochemical characteristics, and a phylogenetic analysis based on 16S rDNA gene sequences reveal that these three strains are Bacillus licheniformis, Bacillus pumilus and Brevibacillus borstelensis respectively. GenBank accessions for these three strains are EF439666-EF439668。

Background Uveal effusion syndrome is uncommon in clinic.To understand the clinical characteristics of uveal effusion syndrome is helpful for rescuing visual acuity of patient.Objective This study was to discuss the diagnosis,classification and surgical outcome of uveal effusion syndrome.Methods This was a descriptive study.The clinical data of 14 eys from 10 patients with uveal effusion syndrome,ineluding ophthalmologic examination,B-scan sonography,ultrasound biomicroscopy (UBM),fundus fluorescence angiography (FFA),indocyanine green angiography (ICGA),surgical treatment and prognosis,were retrospectively analyzed.The follow-up period was 6 months.Results The fundus findings of all impacted eyes showed bullous-shape retinal detachment (RD).B-scan sonography revealed retinal and choroidal detachment.A annular peripheral ciliochoroidal detachment was observed in the cases under the UBM.FFA exhibited leopard spots without any leakage from choroid into the subretinal space.ICGA demonstrated diffusely choroidal granular hyperfluorescence in the very early phase,which presented with an increasing intensity as time lapse until the late phase.Full-thickness sclerectomy was performed on 4 eyes of 2 patients and subscleral sclerectomy was performed in 1 eye of 1 patient,achieving a retinal anatomic reattachment after surgery.All of the patients finished the fellow-up.No recurrence of RD was seen during the followup duration.Conclusions Comprehensive preoperative evaluation,including ophthalmologic ultrasonography,MRI and CT,is crucial for accurate classification of uveal effusion syndrome and determine of proper management strategy.

This study is to explore the reason of "the older, the better" of PCR and itsincrease of flavonoids. We identified the fun- gus isolated from the PCR using microscopic and molecular identification. HPLC method was used to determine the content of 4 fla- vonoids and to clarifythe regularity of them; UV spectrophotometry method was used to determine the total content of flavonoids; reverse thinking was applied to screen the fungus that have close relation to the change of flavonoids. Finally, we have isolated and identified 25 fungusfrom the PCR, which belong to 2 genus and 4 species, including pencillium commune, P. minioluteeum, P. citrinum, Aspergillus flavus and A. niger. The content of flavonoids was increased in the mildew PCR due to A. niger and other fungus. Therefore, "the ol- der, the better" of PCR had its scientific reason that the increase of flavonoids had a close relation of the metabolic activity of A. niger and other fungus.
Citrus ; chemistry ; microbiology ; Flavonoids ; analysis ; Fungi ; isolation & purification

OBJECTIVE: To develop a specific, sensitive, reproducible SPE-HPLC method for the determination of 37 drugs in whole blood. METHODS: With the doxapram as internal standard, Oasis column was used to extract drugs from whole blood. Two kinds of mobile phases were used in this study. Separations were achieved by a LiChrospher 100 RP-C18 (250 mm x 4.0 mm x 5 microm) column kept at 50 degrees C, the DAD detector was set at 230 nm and 250 nm. RESULTS: The limit of detection were 1-30 ng/mL. The method showed excellent linearity and the linear correlation coefficient was > or =0.997 98. The relative standard deviation for between-day and within-day assay were <10%. CONCLUSION The method is effective, simple, reliable and has been used in real cases.
Chromatography, High Pressure Liquid/methods* ; Doxapram/isolation & purification* ; Doxepin/isolation & purification* ; Estazolam/isolation & purification* ; Forensic Medicine ; Humans ; Morphine/isolation & purification* ; Papaverine/isolation & purification* ; Pharmaceutical Preparations/isolation & purification* ; Prazosin/isolation & purification* ; Procaine/isolation & purification* ; Reproducibility of Results ; Sensitivity and Specificity ; Solid Phase Extraction/methods* ; Solvents/chemistry*

Objective To evaluate the efficacy of chemoradiotherapy alone and prognostic factors for locally advanced rectal cancer. Methods The clinical data of 47 patients with locally advanced rectal cancer who were admitted to our hospital and mostly treated with chemoradiotherapy alone from 2003 to 2010 were retrospectively analyzed. Three of the patients received radiotherapy alone. The Kaplan?Meier method was used to estimate overall survival (OS), progression?free survival (PFS), and distant metastasis?free survival ( DMFS ) rates, and the log?rank test was used for survival difference analysis and univariate prognostic analysis. The Cox regression model was used for multivariate prognostic analysis. Results In all patients, the 3?and 5?year OS rates were 53?2% and 33?2%, respectively, while the 3?and 5?year PFS rates were 37% and 31%, respectively. During the follow?up, 15 patients (32%) had local progression with PFS of 1?60 months (median PFS, 14 months);23 patients (49%) had distant metastasis with DMFS of 2?60 months ( median DMFS, 17 months) . Patients treated with high?dose radiotherapy had significantly lower 3?and 5?year local progression rates than patients treated with medium?dose radiotherapy ( 11% vs. 54%;11%vs. 57%;P=0?004). After chemoradiotherapy, 9 patients (19%) had clinical complete response (cCR), and the 3?and 5?year OS and PFS rates in those patients were all 8/9. The univariate analysis indicated that tumor distance from the anus and cCR were influencing factors for prognosis ( P= 0?026;P= 0?000 ) . However, the multivariate analysis showed that cCR was the only influencing factor for survival ( HR=12?24;95% CI, 1?64 ?91?29;P= 0?015 ) . Conclusions Chemoradiotherpay or radiotherapy alone is effective and safe in the treatment of patients with locally advanced rectal cancer who have to give up surgery or have unresectable tumors. High?dose radiotherapy may improve local control rate. Complete response to chemoradiotherapy predicts satisfactory treatment outcomes.

OBJECTIVETo study the clinical features and prognosis of gastrointestinal injury caused by foreign bodies in the upper gastrointestinal tract in children.

METHODSA retrospective analysis was performed for the clinical data of 217 children who were diagnosed with foreign bodies in the upper gastrointestinal tract complicated by gastrointestinal injury by gastroscopy from January 2011 to December 2016, including clinical features, gastroscopic findings, complications, and prognosis.

RESULTSAmong the 217 children, 114 (52.5%) were aged 1-3 years. The most common foreign body was coin (99/217, 45.6%), followed by hard/sharp-edged food (45/217, 20.7%) and metal (35/217, 16.1%). The most common gastrointestinal mucosal injury was ulceration (43.8%), followed by erosion (33.2%). Compared with other foreign bodies, button cells were significantly more likely to cause esophageal perforation (P<0.01). The esophagus was the most commonly injured organ (207/217, 95.4%). Of all the 217 children, 24 (11.1%) experienced infection. The children with perforation caused by foreign bodies had a significantly higher incidence rate of infection than those with ulceration caused by foreign bodies (P=0.003). Of all the 217 children, 204 (94.0%) underwent successful endoscopic removal of foreign bodies. Among these children, 98 were hospitalized due to severe mucosal injury and were given anti-infective therapy, antacids, and supportive care including enteral nutrition through a nasogastric tube and/or parenteral nutrition. Of all the children, 10 left the hospital and were lost to follow-up, and all the other children were improved and discharged.

CONCLUSIONSMost cases of foreign bodies in the upper gastrointestinal tract occur at 1-3 years of age. Coin, hard/sharp-edged food, and metal are the most common foreign bodies. Button cells are more likely to cause esophageal perforation. The incidence rate of secondary infection increases with the increasing severity of gastrointestinal mucosal injury. Children undergoing endoscopic removal of foreign bodies and enteral nutrition through a nasogastric tube tend to have a good prognosis.

Female ; Food ; adverse effects ; Foreign Bodies ; diagnosis ; etiology ; therapy ; Humans ; Infant ; Male ; Metals ; adverse effects ; Prognosis ; Retrospective Studies ; Upper Gastrointestinal Tract ; injuries

OBJECTIVETo compare different expression profiles of all known chemokine receptors in human hepatocellular carcinoma (HCC) cell lines with different metastasis potentials.

METHODSEighteen pairs of chemokine receptor primers were designed using Premier software. Expression profiles of the 18 chemokine receptors on four HCC cell lines of lower to higher potentials of metastasis (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) were analyzed by RT-PCR. Expression of CXCR4 was detected by RT-PCR.

RESULTSExpression profiles of chemokine receptors on four HCC cell lines with different metastatic potentials had significant differences (P < 0.01), in which CCR10, CXCR4 and CXCR6 expressions decreased gradually as the metastatic potential of the cell lines increased. The expressions of CCR3, CCR4, CCR10, CCR12 and XCR1 on HCCLM6 were significantly reduced compared with SMMC-7721 (P < 0.01), whereas the expressions of CXCR1 (P = 0.006) and CXCR5 (P = 0.003) exceeded that of SMMC-7721. Except for CXCR2, CXCR6 and XCR1, most of chemokine receptors on MHCC97-H were expressed differently compared with MHCC97-L (P < 0.05), in which expressions of CCR1 (P = 0.002), CCR2 (P = 0.004) and CCR5 (P = 0.046) exceeded MHCC97-L. CXCR4 was detected only on the positive controls and SMMC-7721 when the template of total RNA was reduced one-half in RT-PCR.

CONCLUSIONChemokine receptors are expressed very differently at mRNA level on HCC cell lines with different metastatic potentials. The different profiles of chemokine receptors in tumor microenvironment and the function of CXCR4 in HCC should be further studied. Our findings have important implications in understanding the relationship between chemokine receptors and the metastatic potential of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Chemokine ; metabolism

A boy aged 14 years had abdominal pain as the major manifestation, with elevated serum amylase and lipase. Abdominal ultrasound performed early after onset in another hospital showed enlargement of the pancreas and a reduction in echo. Magnetic resonance cholangiopancreatography (MRCP) showed pancreatic duct dilation and an unclear image of the head of the pancreas. Acute pancreatitis was considered. However, his symptoms were not relieved after fasting, fluid infusion, anti-acid therapy, and somatostatin therapy. Then, abdominal CT scan and MRCP found multiple low-density lesions of the pancreas and enlargement of the hilar and retroperitoneal lymph nodes. Exploratory laparotomy found pancreatic edema and multiple hilar nodules with unclear boundaries, and pathological biopsy showed anaplastic large-cell lymphoma. Since the liver, the spleen, bone marrow, and the central nervous system were not involved, he was diagnosed with stage III primary pancreatic lymphoma. After vindesine and dexamethasone were used to reduce tumor load, the patient underwent vindesine-pirarubicin-asparaginase-dexamethasone chemotherapy once and vinorelbine-dexamethasone chemotherapy 8 times. Imaging examination still showed multiple low-density lesions of the pancreas and retroperitoneal lymph node enlargement. His parents discontinued treatment. It is concluded that the rare causes of acute pancreatitis with poor response to conventional treatment should be considered, especially for patients with abdominal lymph node enlargement. Extranodal lymphoma should be considered, and lymph node biopsy should be performed as early as possible to confirm diagnosis. The prognosis of pancreatic lymphoma is associated with clinical stage and pathology.