AlM: To explore the different ages of congenital nasolacrimal duct obstruction in infants, take different treatment methods at different times.
METHODS:The 87 cases of 102 children were divided into three different age groups: the first group of 25d-3mo of age 21 cases 26 eyes; The second group >3mo-7mo 31 cases 36 eyes;The third group >7-24mo of age 35 cases 40 eyes. For the first group of infants, the implementation of the lacrimal sac nasolacrimal duct massage + eye drops; for the second group of infants, carry lacrimal pressure washing treatment; for the third group of infants, the implementation of the nasolacrimal duct probing treatment.
RESULTS: The first group of children through the nasolacrimal duct sac massage + drops tobramycin eye drops treatment unobstructed 12, the cure rate was 46. 2%;The second group of children through pressurized irrigation treatment lacrimal patency by 33, the cure rate was 91. 7%; The third group of children through the nasolacrimal duct probing unobstructed 36 treatment, the cure rate was 90. 0%. The second and third group were better than the first group (χ2=15. 71, P<0. 01;χ2=15. 27, P<0. 01);the treatment effect of the second and third groups was no significant difference (χ2=0. 02, P>0. 05).
CONCLUSlON:lnfants with congenital nasolacrimal duct obstruction should distinguish between ages, taking different treatments, in order to obtain a better therapeutic effect, and lacrimal pressure washing is the preferred way of treating infants with congenital nasolacrimal duct obstruction.

Objective To investigate whether a novel long-acting tumor necrotic factor (TNF) antagonist (soluble TNF receptor:IgG Fc [sTNFR:IgG-Fc]) can protect hepatocyte damage against liver failure caused by drugs in immunity-induced cirrhotic rats.Methods Wistar rats were repeatedly sensitized by human serum albumin (HSA) emulsified in complete freud adjuvant.The blood was collected at day 10 after the final sensitization.If anti-albumin antibody was positive,the rats were intravenously injected with HSA twice a week.After six weeks,liver cirrhosis was induced by immunity.All the model rats were divided into three groups with 15 each.Liver failure was induced with D-galactosamine/ lipopolysaccharide (LPS) intraperitoneal injection in the rats with liver cirrhosis in model group.The rats in pretreatment group were intraperitoneally injected with long-acting soluble TNF receptor p55 18 h before D-galactosamine/LPS injection.The control group were injected with 0.9％ sodium chloride.General condition,survival rate,liver function and pathological changes were all examined.Serum levels of interleukin (IL)-6,IL-22 and intrahepatic level of IL-6 were detected by enzyme linked immunosorbent assay (ELISA).The activity of Caspase 3 in hepatocyte lysis solution was measured by spectrophotography.Real-time polymerase chain reaction (PCR) was used to detect mRNA expressions of proliferating cell nuclear antigen (PCNA),bcl-2,bax and IL-22 receptor.Data were analyzed by variance analysis among groups.Results Rats in model group were dispirited with poor response after 12 hours and only 3 survived,compared with soluble TNF receptor p55 pre-treated group rats,in which all survived (P=0.029 8) with flexible response.Serum alanine aminotransferase levels in these two groups were (6 533± 360) and (105 ± 7) U/L,respectively.Hepatic regenerative nodule developed massive or submassive necrosis with septal fibrosis in model group,whereas soluble TNF receptor p55 alleviated the inflammatory and necrosis reaction of hepatic tissue.Serum IL-6 levels in model group and pretreatment group were (842.0±12.9) and (91.9±1.6) pg/mL,respectively (F=380.30,P＜0.01).Intrahepatic levels of IL-6 in these two groups were (26.2±1.2) and (11.1±0.8) pg/mL,respectively (F=176.90,P＜0.01),and serum IL-22 levels were (167.0±27.8) and (988.0±109.6) pg/mL,respectively (F=37.91,P＜0.01).Hepatic Caspase-3 activity was reduced by almost 60％ by soluble TNF receptor p55 pretreatment (F=303.70,P＜0.01) and bax expression reduced by 22％ (F=108.80,P＜0.01),while bcl-2 and PCNA expressions were up-regulated by 3.6-folds and 23.0-folds,respectively (F=115.60,P＜0.01; F=594.20,P＜0.01).Conclusions Long acting soluble TNF receptor p55 could improve survival rate,liver function and reduce inflammatory reaction of rats with liver failure induced by drugs on the basis of liver cirrhosis caused by immunity,which indicates that this drug may process a potential therapeutic value.

Objective To investigate the clinical features of and GJB2 gene mutations in a Chinese Han patient with keratitis-ichthyosis-deafness syndrome (KID syndrome),in hope to offer evidence for the clinical and genetic diagnosis of KID syndrome.Methods Clinical data were collected from a patient with KID syndrome.DNA was extracted from peripheral blood of the patient and his two family members (mother and brother).PCR was performed to amplify the exon 2 and its flanking splicing sites of GJB2 gene followed by bidirectional direct DNA sequencing. Results The patient presented with the typical triad of vascularizing keratitis,ichthyosis and congenital deafness.A G148A mutation in the exon 2 of GJB2 gene,resulting in the substitution of aspartic acid by asparagine at position 50 of the junction protein connexin 26 (Cx26),was identified in the patient,but not in either of his family members.Conclusion The G148A mutation in GJB2 gene may be responsible for the clinical phenotype of KID syndrome in this Chinese patient.

Objective To construct the Neisseria surface protein A (NspA) DNA vaccine of Neisseria gonorrhoeae and evaluate the humoral and cellular immune responses induced by this vaccine in mice model.Methods The recombinant expression vector pcDNA3.1 (+)/NspA was constructed by inserting NspA gene into the eukaryotic expression vector pcDNA3.1 (+) and confirmed by poly merase chain reaction (PCR),restriction enzymes HindⅢ,XbaⅠand DNA sequencing.NspA mR- NA in transfected RAW264.7 cells and NspA protein expression in transfected COS-7 cells were de- tected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical stai- ning,respectively.Forty-five male BALB/c mice were immunized with pcDNA3.1 (+)/NspA recom binant plasmid.The level of serum anti-Neisseria gonorrhoeae antibody of the immunized mice was detected by tube agglutination test,and the level of interieron (IFN)-?was assayed by enzyme-linked immunosorbent assay (ELISA).The proliferation of splenocytes was determined by methyl thiazolyl tetrazolium (MTT) colormetry.The NspA gene in BALB/c mice was identified by PCR with the total DNA extracted from quadriceps femoris in immunized sites.Results Restriction enzymes digestion a- nalysis and DNA sequencing results revealed that the pcDNA3.1 (+)/NspA had been constructed successfully.NspA gene had been transcripted and expressed in mammalian cells.The peak titer of specific antibody was 1:640 in pcDNA3.1(+)/NspA immunized group and there was no specific an- tibody detected in both pcDNA3.1 (+) immunized group and PBS group.The IFN-?level in pcD NA3.1 (+) immunized group was (23.79?11.85)pg/mL and that in pcDNA3.1 (+)/NspA immu- nized group was(169.71?30.52)pg/mL (P

OBJECTIVETo study the effects of anti-oncogene WWOX on cell growth of cholangiocarcinoma.

METHODSThe expression of WWOX protein was detected with immunohistochemical method-SP in 54 patients with cholangiocarcinoma from July 2005 to May 2010 and 12 samples of normal bile duct tissues. The recombinant WWOX eukaryotic expression plasmid was introduced into RBE cells by liposome-mediated transfection and positive cell clones were selected and amplified. The mRNA and protein expressions in RBE cells stably transfected with WWOX were investigated by quantitative RT-PCR and Western Blot before and after transfection. Cell proliferation was tested by MTT, cell apoptosis was assessed by FCM, the alteration of mitochondria membrane potential (ΔΨm) was detected by JC-1 staining method, cell invasion was determined by Transwell chamber assay. The expression change of bcl-2, bax, FasL, caspase-3 mRNA and protein was detected by quantitative RT-PCR and Western Blot.

RESULTSThe expression of WWOX protein was significantly lower in cholangiocarcinoma than that in normal bile duct tissues and loss of WWOX protein expression was found in 40.7% of cholangiocarcinoma specimens (P < 0.05). RBE cells with stable transfection of WWOX were established. Quantitative RT-PCR showed that the expression of WWOX mRNA was significantly enhanced and Western Blot demonstrated that WWOX protein expression was markedly increased. MTT showed that WWOX gene transfection significantly decreased the proliferation of RBE cells (P < 0.05). FCM analysis showed that the apoptosis rate after transfection was significantly promoted [(1.1 ± 0.6)% vs. (1.7 ± 0.5)% vs. (35.2 ± 4.4)%, P < 0.01], JC-1 staining method indicated that the experimental group was loss of ΔΨm [(12.6 ± 1.9)% vs. (13.6 ± 1.8)% vs. (48.7 ± 2.9)%, P < 0.01], transwell chamber assay showed that the number of transfected cells that passed the transwell membrane was significantly less than those of control groups (77 ± 6 vs. 72 ± 8 vs. 48 ± 6, P < 0.01). Quantitative RT-PCR and Western blotting showed that the expression of bcl-2 mRNA and protein was markedly decreased and the expression of bax, caspase-3 were significantly increased. There was no significant change in the expression of FasL.

CONCLUSIONWWOX exerts its antitumor effect against proliferation through inducing cell apoptosis in cholangiocarcinoma.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Cholangiocarcinoma ; genetics ; metabolism ; pathology ; Genetic Vectors ; Humans ; Oxidoreductases ; genetics ; metabolism ; Plasmids ; genetics ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism ; WW Domain-Containing Oxidoreductase

OBJECTIVEImpact of the presence of bacteria associated with a marine dinoflagellate, Alexandrium tamarense CI01, on the growth and toxin production of the algae in batch culture was investigated.

METHODSPronounced changes in the activities of the algal culture were observed when the culture was treated with different doses of a mixture of penicillin and streptomycin.

RESULTSIn the presence of antibiotics at the initial concentration of 100 u/mL in culture medium, both algal growth and toxin yield increased markedly. When the concentration of antibiotics was increased to 500 u/mL, the microalgal growth was inhibited, but resumed in a few days to eventually reach the same level of growth and toxin production as at the lower dose of the antibiotics. When the antibiotics were present at a concentration of 1 000 u/mL, the algal growth was inhibited permanently.

CONCLUSIONSThe results indicate that antibiotics can enhance algal growth and toxin production not only through their inhibition of the growth and hence competition for nutrients, but also through their effects on the physiology of the algae.

Animals ; Anti-Bacterial Agents ; pharmacology ; Bacteria ; Dinoflagellida ; microbiology ; pathogenicity ; Eutrophication ; Marine Toxins ; biosynthesis ; Penicillins ; pharmacology ; Saxitoxin ; Streptomycin ; pharmacology

OBJECTIVETo evaluate the short-term effect and experience of Nuss procedure on 120 cases of patients with pectus excavatum.

METHODSThoracoscopy assisted Nuss procedure with different ways of anesthesia were applied to 120 cases of patients with pectus excavatum, including 7 cases of recurrence after traditional surgical procedure (6 cases) and Nuss method (another one). The patients ranged in age from 2.5 to 43 (mean 14.1) years and in Haller index from 2.91 to 29. Of the 120, 73 had symmetric and 47 had asymmetric pectus excavatum. The Nuss procedure is performed with general anesthesia and a convex steel bar is inserted under the sternum with thoracoscopy through small bilateral thoracic incisions. The steel bar is inserted with the convexity facing posteriorly, and when it is in position, the bar is turned over, thereby correcting the deformity.

RESULTSThe operation was successfully accomplished without severe complications in all the 120 cases. The mean operative time was 58 minutes and the mean volume of blood loss was 30 ml. 103 patients had one bar inserted while the other 17 cases with more extremely diffuse depression required 2 or even 3 bars to get a satisfactory correction. Such methods as modifications to the fixing points and the shape of the bar, partial osteotomy, were developed to deal with asymmetric ones.

CONCLUSIONThe Nuss procedure is a minimally invasive technique for correction of pectus excavatum. It can lead to a satisfactory outcome and surgical time is less.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Funnel Chest ; surgery ; Humans ; Male ; Orthopedic Procedures ; methods ; Thoracoscopy ; Young Adult

OBJECTIVETo survey an outbreak of acute gastroenteritis in Lulong County and analyze the cause of the disease.

METHODSEpidemiological methods were applied to investigate an outbreak of acute gastroenteritis occurred in June 2000 in Lulong County. Stool specimens were collected from diarrhea patients and were tested for human calicivirus by ELISA and RT-PCR. The products of RT-PCR were cloned and sequenced, then phylogenetic analysis was carried out.

RESULTSIn total, 736 farmers were surveyed, among them 134 had acute gastroenteritis, the attack rate was 18.20%, and one elderly patient died. The age of patients was from 1 to 77 years and the incidence of the disease among young people was higher with a peak in June 25 through 30. Six stool specimens were tested for caliciviruses by ELISA and 3 were positives, one of them was confirmed by RT-PCR and belonged to norovirus genotype GI/2. No other pathogens were detected.

CONCLUSIONHuman calicivirus was confirmed to be the cause of the outbreak of acute gastroenteritis.

Acute Disease ; Caliciviridae ; genetics ; isolation & purification ; Caliciviridae Infections ; epidemiology ; China ; epidemiology ; Disease Outbreaks ; Enzyme-Linked Immunosorbent Assay ; Feces ; virology ; Gastroenteritis ; epidemiology ; virology ; Humans ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

To study the enzyme kinetics of schizandrin metabolism in different gender in rat liver microsomes, liver microsomes were prepared from male or female rats. Schizandrin was incubated with rat liver microsomes. Schizandrin and its metabolites were isolated and identified by HPLC-UV method. Vmax, Km and Cl(int) of schizandrin in male and female rat liver microsomes were (21.88 +/- 2.30) and (0.61 +/- 0.07) micromol x L(-1) x min(-1) x mg(-1) (protein), (389.00 +/- 46.26) and (72.64 +/- 13.61) micromol x L(-1), (0.0563 +/- 0.0007) and (0.0084 +/- 0.0008) min x mg(-1) (protein), respectively. The major metabolites of schizandrin in female and male rat liver microsomes were 7,8-dihydroxy-schizandrin (M1) and 7, 8-dihydroxy-2-demethyl schizandrin (M2b), respectively. Ketoconazole, quinidine, and orphenadrine had different level effects on schizandrin metabolism in both male and female rat liver microsomes, and cimetidine still had some inhibitory effect in male liver microsomes. CYP3A and CYP2C11 may be the main P450 enzymes in schizandrin metabolism and their difference in rat liver microsomes may be the main reason for the sex difference of metabolic enzyme kinetics and metabolites of schizandrin in rats.
Animals ; Chromatography, High Pressure Liquid ; Cimetidine ; pharmacology ; Cyclooctanes ; isolation & purification ; metabolism ; Enzyme Inhibitors ; pharmacology ; Female ; In Vitro Techniques ; Ketoconazole ; pharmacology ; Lignans ; isolation & purification ; metabolism ; Male ; Microsomes, Liver ; metabolism ; Orphenadrine ; pharmacology ; Plants, Medicinal ; chemistry ; Polycyclic Compounds ; isolation & purification ; metabolism ; Rats ; Rats, Sprague-Dawley ; Schisandra ; chemistry ; Sex Factors ; Spectrophotometry, Ultraviolet

The aim of this paper is to develop and validate a rapid and sensitive LC-APCI/MS method for the determination of triptolide (TP) in plasma and to study the pharmacokinetic properties of TP in Beagle dogs. Sample preparation consisted of liquid-liquid extraction of interests. with ethyl acetate from dog plasma. The analytes and internal standard prednisolone were well separated on a Zorbax Extend-C18 analytical column. Plasma TP was detected by selected-ion monitoring (SIM) of LC-APCI/MS as its deprotonated molecular ions [M - H] - at m/z 358.9. Pharmacokinetic studies were undertaken in dogs following an iv dose of 0.05 mg x kg(-1) of TP or an ig dose of 0.05, 0.08, 0.1 mg x kg(-1), separately. The pharmacokinetic parameters were calculated by DAS software. Calibration curves were linear over the concentration range of 1 - 200 ng x mL(-1) of TP with the within- and between-batch precisions less than 10%. The within and between-batch accuracy was 95.0% to 105.0%. Recovery of LC-MS method for TP in plasma was over 75%. The T1/2beta was (2.5 +/- 0.8) h after intravenous administration of TP at the dose of 0.05 mg x kg(-1). There were no significant differences in T(max), T1/2 alpha and T1/2 beta among the three ig dosage groups. AUC and C(max) increased proportionally with doses. The absolute bioavailability of TP after ig administration of 0.05 mg x kg(-1) was (75 +/- 17)%. The LC-MS method for determination of triptolide in dog plasma was sensitive and rapid. It was showed that the elimination of triptolide was rapid. The absolute bioavailability of triptolide given orally was high.
Administration, Oral ; Animals ; Area Under Curve ; Biological Availability ; Chromatography, High Pressure Liquid ; Diterpenes ; administration & dosage ; blood ; pharmacokinetics ; Dogs ; Dose-Response Relationship, Drug ; Epoxy Compounds ; administration & dosage ; blood ; pharmacokinetics ; Female ; Injections, Intravenous ; Male ; Mass Spectrometry ; Phenanthrenes ; administration & dosage ; blood ; pharmacokinetics ; Plants, Medicinal ; chemistry ; Random Allocation ; Tripterygium ; chemistry