AlM: To observe the early tear film changes after trabeculectomy using Keratograph 5M.METHODS:Fourty-one patients (46 eyes) of glaucoma who underwent trabeculectomy were involved. First tear break up time ( FTBUT ) , average tear break up time (ATBUT), non-invasive tear meniscus height (NlTMH), lipid layer thickness and meibomian gland scores were measured at 1d preoperatively and at 1d, 1wk, 1mo postoperatively.RESULTS:At 1d, 1wk and 1mo postoperatively, FTBUT and ATBUT decreased greatly (P<0. 05), as well as lipid layer thickness (P<0. 017). NlTMH at 1d postoperatively increased significantly, but at 1wk and 1mo postoperatively decreased greatly (P<0. 05). There was no significantly difference in meibomian gland scores between preoperation and postoperation (P>0. 05). CONCLUSlON: Keratograph 5M could be used to evaluate tear film function rapidly and accurately. Trabeculectomy significantly alters tear film stability and tear secretion in the short term after operation.

Objective To explore the neuropsychological development in twin premature infants,and to analyze the risk factors for the retardation of development.Methods The premature infants hospitalized in the Neonatal Ward First Affiliated Hospital of Kunming Medical University from Jun.2010 to Jun.2012 were divided into study groups of twin premature infants and a control group of singleton premature infants.The neuropsychological development of 88 premature infants was evaluated at corrected gestational age of 1 year old by Bayley scales of infant development.According to normal lab findings of the Bayley scales of infant development,the premature infants were then divided into abnormal neuropsychological development group [mental development index(MDI) ≤79 scores] and normal neuropsychological development group (MDI ＞ 79 scores),and the data of 2 groups were statistically analyzed.Results The frequency distribution of MDI score in study group showed a skewed distribution,of which 59％ (27/46 cases) had abnormal neuropsychological development (MDI ≤ 79 scores); However,the frequency distribution of MDI score in control group showed an approximate normal distribution,of which only 12％ (5/42 cases) for abnormal neuropsychological development.Head circumference,body length,body weight,MDI,psycho-motor development index of twin premature infants of one-year old were lower than those of the singleton premature infants of the same age,and the differences were statistically significant (all P ＜0.05).The single factor analysis demonstrated that the risk factors for abnormal neuropsychological development (MDI ≤ 79 scores) of twin premature infants were gestational age,birth weight,mother' s gcstational age,the cultural degree of parents,mode of feeding,neonatal hyperbilirubinemia,neonatal hypoglycemia and neonatal sepsis.Multiple regression analysis showed that neonatal hyperbilirubinemia was the independent risk factor for abnormal neuropsychological development of twin premature infants.Conclusions At same gestational age,neuropsychological development of twin premature infants lagged behind singleton premature infants.Neonatal hyperbilirubinemia may be the risk factors for neuropsychological development of twin premature infants,so the earlier management of neonatal hyperbilirubinemia may be beneficial for the neuropsychological development of twin premature infants.

Evaluation Studies as Topic ; Humans ; Medicine, Chinese Traditional ; Publishing ; standards ; Quality Control ; Randomized Controlled Trials as Topic ; standards

0.05).The results of electrocardiography and the laboratory ex-amination showed that neither ANWin cluding natural Calculus Bovis nor A NWincluding in -vitro cultured Calc ulus Bo-vis had obviously toxic and side effe cts in treating epidemic encephalitis B.Conclusion ANW including in -vitro cul-tured Calculus Bovis has an markedly effect in the treatment of epidemic e ncephalitis B.

AIM: To investigate the effect of venous marker chicken ovalbumin upstream promoter-transcription factor Ⅱ (COUP-TFⅡ) expression on vascular endothelial cell senescence and its molecular mechanism.METHODS: The mRNA expression of COUP-TFⅡ in the human coronary artery endothelial cells (HCAEC) and human umbilical vein endothelial cells (HUVEC) was detected by RT-qPCR.After transfection with a COUP-TFⅡ siRNA (siCOUP-TFⅡ) to inhibit COUP-TFⅡ expression in the HUVEC, the senescence and proliferation of endothelial cells were evaluated by β-galactosidase staining, Western blot, CCK-8 assay and cell counting after treatment with 10-5 mol/L angiotensin Ⅱ (AngⅡ).The protein levels of Akt and p-Akt were determined by Western blot.RESULTS: Compared with the HCAEC, COUP-TFⅡ was significantly highly expressed in the HUVEC.Knockdown of COUP-TFⅡ via siCOUP-TFⅡ significantly induced endothelial cell senescence and inhibited endothelial cell proliferation and p-Akt level after treatment with AngⅡ at 10-5 mol/L.Furthermore, an Akt activator SC79 at 4 mg/L partly reversed the effect of siCOUP-TFⅡ on AngⅡ-induced endothelial cell senescence and proliferation.CONCLUSION: Knockdown of COUP-TFⅡ promotes endothelial cell senescence and inhibits endothelial cell proliferation, which might be partly regulated by Akt signaling.

OBJECTIVETo observe the regulating effect of hepatitis C virus (HCV) envelop (E) 2 gene immunization-induced immune responses by adenovirus mediated interleukin 12 (IL-12).

METHODSHCV E2 protein was expressed and purified from NIH 3T3 and then used as an antigen to detect antibodies against HCV E2. With 51Cr release, SP2/0 expressing HCV E2 was used as target cell to detect specific cytotoxic T lymphocytes (CTL) response; adenovirus recombined IL-12 was propagated by 293 cell. HCV E2 recombinant and adenovirus recombined IL-12 were injected into the quadriceps femoris muscles and abdominal cavities of 6-8 weeks old BALB/C mice. Sera were collected at 2, 3, and 4 weeks and detected for antibodies for E2. Spleen cells isolated at 4 weeks were analyzed for specific CTL response.

RESULTSIt was found that expression of IL-12 at an undetectable level did enhance HCV E2 gene immunization-induced CTL activity and there was no effect on its hormonal immune response.

CONCLUSIONUsing adenovirus to express interleukin 12 was helpful for regulation of HCV E2 gene immunization-induced immune response. Combined HCV E2 and IL-12 can render a strong anti-HCV CTL activity and may be of use in the development of HCV gene vaccine in the future.

Adenoviridae ; physiology ; Interleukin-12 ; biosynthesis ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; Viral Envelope Proteins ; genetics ; immunology

AIMTo evaluate the hypoglycemic effect of chitosan-coated and sodium alginate-coated insulin liposomes after oral administration in mice.

METHODSInsulin-liposomes were prepared by reverse-phase evaporation. Chitosan and alginate coating was carried out by mixing liposomal suspension with chitosan and sodium alginate solutions, followed by incubation. The particle size and morphology of insulin-liposomes were determined using laser light scattering instrument and transmission electron microscopy (TEM). The entrapment efficiency was analyzed using HPLC and ultracentrifuge. The protection of insulin from peptic and tryptic digestion was studied with HPLC. The hypoglycemic effects of polysaccharide-coated insulin liposomes were investigated using the glucose oxidase method after oral administration in mice.

RESULTSThe particle size of uncoated, chitosan-coated and alginate-coated insulin-liposomes was (138 +/- 31) nm, (230 +/- 20) nm and (266 +/- 19) nm, respectively. All insulin-liposomes were of spherical or ellipsoidal shape. The entrapment efficiencies were 81.6%, 73.5% and 68.7%, respectively. Insulin was protected from tryptic digestion by chitosan-coated liposomes and protected from peptic digestion by alginate-coated liposomes. The hypoglycemic effects of insulin-liposomes, coated with 0.1% chitosan and 0.1% sodium alginate, were observed.

CONCLUSIONChitosan-coated and sodium alginate-coated liposomes were shown to reduce peptic or tryptic digestion on insulin, and enhance enteral absorption of insulin.

Administration, Oral ; Alginates ; Animals ; Blood Glucose ; metabolism ; Chitin ; analogs & derivatives ; chemistry ; Chitosan ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Glucuronic Acid ; Hexuronic Acids ; Hypoglycemic Agents ; administration & dosage ; pharmacology ; Insulin ; administration & dosage ; pharmacology ; Liposomes ; Male ; Mice ; Particle Size ; Random Allocation ; Technology, Pharmaceutical ; methods

Objective To discuss the surgical procedures and treatment after combined liver and intestinal transplantation with portal venous drainage and enterostomy of two ends in one case.Methods A male patient with liver dysfunction and short bowel syndrome underwent the combined liver and intestinal transplantation.With the techniques of en bloc,the liver and intestinal grafts were harvested from cadaveric donor.The intestinal graft,200 cm long,was implanted with portal venous drainage and aortic inflow,and enterostomy of both ends was performed instead of intestinal anastomosis.The liver graft was placed in a piggyback fashion with end to end anastomosis of the bile ducts without T tube. Inmunosuppression protocol was administrated with campath-1H and tacrolimus.Endoscopic biopsy of intestinal graft was performed regularly,and clinical observation was done to monitor the acute rejection.Results In the first month after operation,abdominal infection was controlled by intraperitoneal drainage with open surgery.One suspect acute rejection was treated with methylprednisolone.Until sixth month,the functions of liver and intestine were progressively restored.However,the patient lost weight and could not be free from intravenous nutrition because of diarrhea.Conclusion Combined liver and intestinal transplantation with portal venous drainage and enterostomy of two ends is a simple surgical procedure with lower risk of surgical complications.This method is propitious to monitoring rejection and function improvement of the grafts.Diarrhea and loss of digestive juice are the main reasons of low body weight and malnutrition.

AIMTo study the transfection and anti-hepatitis B virus (HBV) effect of the co-modified hepatocytes-targeting cationic liposomes encapsulating anti-HBV antisense oligonucleotides (asON) , and to investigate the transfection mechanisms of the liposomes.

METHODSDipalmitoylphosphatidylcholine (DPPC) and 3beta-[N-(N',N'-dimethylaminoethane)-carbamoyl] cholesterol (DC-Chol) were used as the lipids, beta-sitosterol-beta-D-glucoside (sito-G) and Brij 35 were used to modify the liposomes. Flow cytometry (FCM), fluorescence microscopy and enzyme-linked immunosorbent assay (ELISA) were utilized to evaluate the transfection improvement of the asON encapsulated in the liposomes in primary rat hepatocytes and the antigens inhibition activity in HepG 2.2.15 cells. The transfection mechanisms were evaluated based on the influence of wortmannin, nigericin, and asialofetuin on the antigens inhibition in HepG 2.2.15 cells by ELISA.

RESULTSThe co-modification with sito-G and Brij 35 significantly improved the transfection of the liposomes in primary rat hepatocytes and antigens inhibition effect in HepG 2.2.15 cells. Both transfection efficiency and antigens inhibition effect showed to be concentration-dependent with the asON-encapsulating liposomes. In fluorescence microscopy, the transfected cells showed strong fluorescence in primary rat hepatocytes, especially in the nuclei. Wortmannin, nigericin and asialofetuin decreased the antigens inhibition of the asON-encapsulating liposomes to different levels. Cationic liposomes modification with sito-G and Brij 35 could improve the transfection and antigens inhibition effect of the asON. The transfection mechanisms of the co-modified liposomes included endocytosis and membrane fusion. The ligand sito-G was confirmed to be able to enhance asialoglycoprotein receptor (ASGPR)-mediated endocytosis.

CONCLUSIONCo-modified hepatocytes-targeting cationic liposomes would be a specific and effective carrier to transfer asON into hepatocytes.

Androstadienes ; pharmacology ; Animals ; Asialoglycoproteins ; pharmacology ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cell Survival ; Cells, Cultured ; Endocytosis ; drug effects ; Female ; Fetuins ; Flow Cytometry ; Hepatitis B Antigens ; metabolism ; Hepatitis B virus ; genetics ; immunology ; Hepatocytes ; cytology ; drug effects ; metabolism ; Humans ; Liposomes ; Microscopy, Fluorescence ; Nigericin ; pharmacology ; Oligonucleotides, Antisense ; chemistry ; genetics ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Wistar ; Sitosterols ; chemistry ; Transfection ; methods ; alpha-Fetoproteins ; pharmacology

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C