Objective To approach the method for improving the intellectual development,of whom had suffered from parity′s brain damage.Methods The patients were divided into 2 groups randomly(age range from 7 days to 18 months) who were observed and compared for 3 years.Group A was bundle of intervention which had been under early intervention program since infant.Group B was bundle of comparing of which had not been under early intervention.Results Observed for 6 months and 12 months,the intellectual development of group A was prior to that of group B(age 6 months P

Acute Disease ; Adult ; Central Nervous System Diseases ; diagnosis ; etiology ; Decompression Sickness ; complications ; diagnosis ; Humans ; Magnetic Resonance Imaging ; Male ; Retrospective Studies ; Young Adult

Objective To establish a simple and sensitive method for the determination of serum copper by spectrophotometry.Methods Nitro-PAPS was used as a coloring agent for serum copper in the presence of surfactants Tween-80 and Triton X-100 and the formed complex was measured by spectrophotometry.Results The maximum absorption wavelength of the complex was 570 nm and the molar absorption coefficient was 7.95?10~4 L/(mol?cm).The lineafity of the method was up to 63.0 ?mol/L and the recoveries ranged from 98.6% to 103.1%.The within-run and between-run CVs were 2.1%-3.3% and 2.7%-3.8%.The method(Y)was compared with an AAS method(X)and a correlation of Y=1.01X -0.27(r=0.998 2)was obtained.A reference interval(x~-?2s)determined with this method on 68 individuals was 9.7-24.1 ?moL/L.Conclusions A simple and sensitive method for serum copper has been established.It may used for the analysis of serum copper in clinical laboratories.

Many bioactive peptides from neural and endocrine tissue are amidated at C-terminals,which is essential for their activities.The ?-amide comes from post-translational modification that is catalyzed by ?-AE (?-amidating enzyme) or PAM (pepdilylglycine ?-amidating monooxygenase).The gene encoding ?-AE was amplified with PCR and cloned into the plasmid pET-30a.After the recombinant plasmid pET-A was transformed into E.coli BL21,the ?-AE was expressed and purified by the Ni2+affinity chromatography,which has the ability catalyzing Dns-Tyr-Val-Gly to Dns-Tyr-Val-NH2.It identified that the recombinant protein producing by E.coli BL21 is ?-AE,which will benefit for studies of amidation at the C-terminals of peptides.

To obtain a number of penicillinases and degrade penicillin in milk by using the penicillinases,the gene encoding penicillinase was amplified by PCR from Bacillus cereus ATCC10987,cloned into pET28a(+) ,transformed into E. coli BL21;analysis of SDS-PAGE and penicillinase activity of the recombinant protein were done under induction of IPTG and the result showed that the maximum penicillinase activity reached 480 U/mL;the purity of penicillinase purified by Ni2+ Purification System was more than 90%;the immobilized penicillinases were obtained by sodium periodate method and the residual quantity of penicillin in milk(containing 0.5 U penicillin G/mL) was less than 4 ppb after degraded by the immobilized penicillinase.

SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.

Objective　 To investigated the possible roles of IL-18 in HBV infection, and the expression of IL-18 in the peripheral blood mononuclear cells (PBMC) of chronic hepatitis B. Methods 　In 15 cases of asymptomatic carriers, 30 cases of active and remissive phases of chronic hepatitis and 10 healthy individuals (as normal controls), IL-18 expression in PBMC were quantitatively analyzed with flow cytometric immunological method. The PBMC were separated routinely and stimulated with LPS (SIGMA) and Monensin (SIGMA) for 6 hours. Then the cells were harvested and fixed by PBS/4% paraformaldehyde. The treated cells were stored in liquid nitrogen for detection later. After immunologic stain, the expression of IL-18 in PBMC were examined by flow cytometry. Results　 ① IL-18 was the lowest in asymptomatic carriers. It was fewer in remissive phase than in normal controls (P＜0.01).There was no difference between active phase and normal controls(P=0.25).②Within the group of active phase, IL-18 was significantly different between each grade of inflammatory activity(P＜0.01),and correlated with serum ALT positively(r=0.63,P＜0.01). Conclusion　 IL-18 may relate to disease activity and the inflammation of liver.

Objective To investigate rite effects of loag-term exposure to low-level sevoflurane on reproductive function in mice.Method F0ny male ICR mice,aged 60 d,weighlag 20-25 g,were randomly divided into 4 groups(n=10 each):control group received no sevoflunme(C);group S1-3 were exposed to 0.003%.0.01% and 0.03% sevoflurane 2 h per day for 5 consecutive days per week for 8 weeks respectively. The mice were then sacrificed at the end of the 8 weeks.The testes and epididymis were emoved and sampled for determination of the activities of total lactic dehydregenase(LDH)and lactic dehydrogenase-X(LDH-X),and the motility rate,amount,and aberration rate of sperm.Testicular uhrastructure were observed by transmission electron microscopy.Results The sperm motifity nne were significantly lower.the sperm aberration rate higher and the activity of LDH-X lower in group S3 than in group C(P<0.05),but there was no significant difference in the above parametem between group SI and group S2(P>0.05).The pathology changed of testes occurred only in group S3 among the 3 groups.Conclusion Long-term exposure to 0.03% sevoflurane can result in the abnormality of the reproductive function in male mice but exposure to≤0.01%sevoflurane dose not.

The rate of corneal graft rejection is still high for high-risk keratoplasty although immune suppression drug is routinely used.The role of traditional Chinese medicine in corneal transplantation is concerned gradually.Heijingpaichitang on the prevention and treatment of rats with high-risk corneal allograft rejection needs further study.Objective This study was to investigate the inhibitory effect of heijingpaichitang on high-risk corneal transplantation immune rejection in rats.Methods Sixteen female SD rats were used as the donors and 32female Wistar rats were served as recipients.The high-risk corneal trasplantation models were established by corneal suture in 32 Wistar rats,and then homogeneity variant SD-Wistar corneal transplantation was performed.The recipients were randomized into model control group,cyclosporinc A (CsA)group,heijingpaichitang group and CsA +heijingpaichitang group.CsA,heijingpaichitang and CsA + heijingpaichitang was orally administered 4 days after operation once per day for 15 days,and normal saline solution was used at the same way in the model control group.Ocular anterior segment reaction was examined under the slit lamp and corneal opacification,edema and neovasculation were scored based on Larkin' s criteria.Rejection index of the corneal graft was recorded and the graft survival time was calculated.The pathological examination of the corneal graft was carried out in all rats,and the inflammatory cells in the corneas and CD4+ cells in the periphery blood were assayed using flow cytometry.The use of the animals complied with ARVO Statement.Results Corneal graft rejection occurred in 10 days after operation in the model control group,12-13 days in the CsA group and heijingpaichitang group and 22 days in the CsA +heijingpaichitang group.Compared with model control group,the scores of the corneal opacification,corneal edema and neovascularization were significantly lower in the CsA group,heijingpaichitang group and CsA+heijingpaichitang group (P＜0.05),and all the scores were declined in the CsA+ heijingpaichitang group compared with CsA group and heijingpaichitang group(P＜0.01),but no significant differences were seen in the scores between the CsA group and heijingpaichitang group(P＞0.05).The mean survival time of grafts was (10.38 ±1.69)days in the model control group,(22.50 ± 3.07) days in the CsA + heijingpaichitang group,with the significant difference (t =-9.790,P =0.000).The pathological examination of graft showed that the lymphocytes and new blood vessels were less in the CsA+heijingpaichitang group compared with CsA group and heijingpaichitang group 15 days after operation.Flow cytometry verified that the number of lymphocytes in graft,CD4+cells and CD4+/CD8+ in periphery blood were significantly lower in the heijingpaichitang group,CsA group and CsA+heijingpaichitang group compared with model control group (P＜0.05).Conclusions Heijingpaichitang can inhibit immune rejection to certain extent in high-risk corneal transplantation rat and has a similar effect to 0.1％ CsA.Heijingpaichitang and 0.1％ CsA have a synergistic effect.

Objective To evaluate the neuroprotective effect and safety of neonatal hypoxic-ischemic encephalopathy(HIE)treated with recombinant human erythropoietin(rhEPO).Methods Fifty-three neonates with HIE were randomly divided into rhEPO treated group(n=29) with the dosage of 300 U/(kg?time),three times a week for 2 weeks and control group(n=24)without rhEPO.All supportive measures were same between 2 groups.Neurological scoring was evaluated at d3,d5 and d7 Neonatal behavioral neurological assessment(NBNA) was evaluated at d7,d14 and d28.The neurodevelopment quote was evaluated at age of 3 and 6 months.Blood pressure,liver and renal function,blood electrolytes and blood hemoglobin,platelet and reticular red blood cell count were monitored before and after treatment in all infants.Results The neurological scoring between two groups had no difference at d3.The significant difference was found at d7(P0.05).Conclusions Teraphy with rhEPO on neonatal HIE infants can promote neurological recovery,and there is no serious side effect with rhEPO treatment.