The cytolytic effect of perforin is a mechanism of anti-virus,killing microbial-infected cells and tumor cells.Perforin is a very important non-specific immune factors in fish.In order to understand the function of perforin,the cDNA of grass carp perforin C-terminal peptide was amplified from grass carp liver and kidney cDNA library.It contains a protein kinase C conserved region 2(C2).The cDNA was connected with pET32a,and transformed to expression bacteria DE3.PFP-C was expressed by a prokaryotic expression system and then purified by affinity chromatography.It showed a significant haemolytic activity when tested with rabbit red cells,the optimal pH for haemolytic activity was 7.5,and its haemolytic function dependents on Ca2+ apparently.

Objective To study the role of activated microglia transplantation in recovery of hindlimb locomotor function in rats with spinal cord injury(SCI). Methods A total of 40 female adult Wistar rats were selected and divided into four groups randomly(10 rats in each group).The former two groups were locomotor function observation groups,in which rat model with spinal cord was established by striking with improved self-made Allen's strike equipment to fabricate moderate spinal cord injury and divided into transplantation group and control group.The latter two were histological observation groups,the spinal cord injury model was fabricated by the same above-mentioned method and divided into transplantation and control groups.Before fabricating the spinal cord injury model,the microglia of the newborn rats were cultured,separated,purified and identified and the purity of the microglia determined.The injury position was exposed again seven days after transplantation and the cell suspension of microglia was injected around the injury position with microsyringe,which was free in the control group.The hindlimb locomotor function of rats was detected and scored at 1 day,1,2,3 and 4 weeks in the locomotor function observation groups after transplantation respectively.At the same time,two rats were extracted randomly from the control group and the transplantation group in histological observation groups to cut specimen and slice for Naoumenko-Feign paraffin section and dying.Then,the microglia were observed and counted by microscope and analyzed statistically. Results At 1,2,3 and 4 weeks after operation,the BBB score of the control group and the transplantation group was increased gradually with the time.But compared with the control group,transplantation group had higher scores of hindlimb locomotor function at 2,3 and 4 weeks after operation,with statistical difference(P＜0.05).Naoumenko-Feigin paraffin section and dying and microglia counting showed that positive microglia number in the transplantation group was increased more obviously than the control group,with statistical difference(P＜0.05).Conclusion Activated microglia transplantation can promote the recovery of the hindlimb locomotor function in SCI rats.

OBJECTIVETo study a new method for the evaluation of the stability of production process of Traditional Chinese Medicines (TCM) based on chromatographic fingerprinting.

METHODTaking the extraction process of Red Ginseng for example, the fingerprints of extracts produced with fluctuating processing factors, i.e. ratio of solvent to liquid, solvent concentration, extraction durations and extraction times, were acquired. Meanwhile, the similarity of fingerprinting based on Euclidean distance was calculated and each fingerprint was fitwith cubic spline.

RESULTSimilarities of fingerprints of the extracts responded to the fluctuation of processing factors, and it showed that taking similarity as index, quantitative relation between chromatographic fingerprints and processing factors could be established.

CONCLUSIONThe method proposed in the paper was able to describe the quality of products resulting from fluctuating conditions in extracting process, and therefore, can be developed as a practicable strategy to evaluate the stability Traditional Chinese Medicines in production process.

Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; isolation & purification ; Evaluation Studies as Topic ; Ginsenosides ; isolation & purification ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Solvents ; Technology, Pharmaceutical ; methods

BACKGROUND: Chitosan is one of the most significant non-viral vector materials with the advantages of outstanding biocompatibility. Quarternary chitosan derivatives can improve transfection efficiency and solubility of chitosan in a broader range of pH values. OBJECTIVE: To synthesize a new vector of quarternary chitosan and to study its complex conditions with plasmid and transfection efficiency compared with chitosan. DESIGN, TIME AND SETTING: A contrast observational study was performed in Second Affiliated Hospital of School of Medicine of Zhejiang University between August and October 2008. MATERIALS: Quarternary chitosan was synthesized in Polymer Materials Lab of Beijing Institute of Technology. Plasmid pEGFP-C1 was presented friendly by Mr. Zheng of School of Medicine of Zhejiang University. Hela cells were provided by Miss. Cheng of School of Medicine of Zhejiang University. METHODS: Quarternary chitosan was prepared according to mass concentration of 0.2 g/L, pH value 5.5 (or 6.9, 7.6) and sodium acetate concentration of 50 mmol/L, and rapidly mixed with pEGFP-C1. The mixture was swirled for 15-30 second and stood at room temperature for 30 minutes at least. MAIN OUTCOME MEASURES: The impacts of pH values and time on complex ability of quarternary chitosan and plasmid were studied by gel retardation test. Transfection efficiency of quarternary chitosan on Hela cells was observed by inversed fluorescence microscope and also compared with chitosan. RESULTS: Quarternaty chitosan could form complex with plasmid in acidic, neutral and basic conditions. It could be used in a broader range of pH values. In an acidic condition, the combination of quarternary chitosan with plasmid was superior to chitosan. A stable complex was formed via a combination of quarternary chitosan or chitosan with plasmid within 30 minutes, and the stability lasted for 12 hours. Transfection efficiency of quarternary chitosan on Hela cells demonstrated that transfection efficiency of quarternary chitosan was superior to chitosan. CONCLUSION: Quarternary chitosan has a broader range in use and higher transfection efficiency than chitosan; however, there is no significant difference in stability between quarternary chitosan and chitosan. Additionally, transfection efficiency of quarternary chitosan on Hela cells is superior to chitosan, which needs a further research.

Aim To iuvestigate the effect and the mechanism of salmonmilt DNA (SMD) on age-related involutions in mouse thymus. MethodsFemale BALB/c of 10 months were divided randomly into three groupsaccording to their weights: high dosage group 333.33 mg/(kg @ d), lowdosage group 166. 67 mg/(kg @ d) and control group 0 mg/(kg @ d) .After five weeks, with Image-Pro Plus (version. 4.0) software, the thymusindexes and the thymoctytes in the thymus section were measured, as wellas the thymus cortex thickness. All the data were analyzed by SAS statisticsoftware. Mieroarray technique was applied to screen the gene fragments,which were differently expressed between the high dosage group and thecontrol group, together with RT-PCR to further confirm some of them.Results No significant differences of the variables including bodyweight, thymus weight and thymus indexes among the three groups werefound (F ＜ 3.0 and P ＞ 0.05, respectively). The thymocytes quanti-ties of thymus cortex and medulla in the high dosage group were significantlyhigher than those of the control group [cortex D(H, C) = 9.46, P ＜ 0.01;medulla t( H.C) = 2.53, P ＜ 0.05]. The thymus cortex thicknesses of bothSMD supplement groups were significantly higher than that of the control group[cortex D(L,C)=3.65, P＞ 0.05; medulla t(L, C)=0.8, P＞ 0.05] .112differently expressed gene fragments were isolated. Furthermore, we foundthe fragments with the logged number of U23789, X80232 and Aw209102were highly expressed in the high dosage group when RT-PCR techniquewas used. Conclusion SMD may reverse the age-related involutions inmouse thymus via up-regulation the expression of proliferation related genesand development and differentiation related genes simultaneously.

Objective:To establish a GC method for the determination of phenol and L-menthol in glycerin Zhiyang lotions. Meth-ods:A Zebron ZB-WAX(0. 32 mm × 30. 0 m,0. 50 μm) capillary column was used with an FID detector. The column temperature was 60℃, maintained for 1 min, and then raised to 160℃ at the rate of 8℃·min-1 , and maintained 10 minutes. The inlet tempera-ture was 180℃, the detector temperature was 300℃, and the carrier gas was nitrogen. Results:The linear range of phenol and L-men-thol was 0. 5-10. 0 mg·ml-1(r=0. 999 9) and 0. 25-5. 0 mg·ml-1(r=0. 999 9), respectively. The average recovery of phenol and L-menthol was 99. 01%(RSD=0. 90%,n=9)and 99. 70%(RSD=0. 98%,n=9), respectively. Conclusion: The method is sim-ple, accurate and reliable, and can be used to determine the concentration of phenol and L-menthol in glycerin Zhiyang lotions.

Background: Glove reuse poses risks, as chemicals can persist even after cleaning. Decontamination methods like thermal aeration, recommended by US OSHA, vary in effectiveness. Some studies show promising results, while others emphasize the importance of considering both permeation and tensile strength changes. This research advocates for informed glove reuse, emphasizing optimal thermal aeration temperatures and providing evidence to guide users in maintaining protection efficiency. Methods: The investigation evaluated Neoprene and Nitrile gloves (22 mils). Permeation tests with toluene and acetone adhered to American Society for Testing Materials (ASTM) F739 standards. Decontamination optimization involved aeration at various temperatures. The experiment proceeded with a maximum of 22 re-exposure cycles. Tensile strength and elongation were assessed following ASTM D 412 protocols. Breakthrough time differences were statistically analyzed using t-test and ANOVA. Results: At room temperature, glove residuals decreased, and standardized breakthrough time (SBT)2 was significantly lower than SBT1, indicating reduced protection. Higher temperature decontamination accelerated residual removal, with ΔSBT (SBT2/SBT1) exceeding 100%, signifying restored protection. Tensile tests showed stable neoprene properties postdecontamination. Results underscore thermal aeration's efficacy for gloves reuse, emphasizing temperature's pivotal role. Findings recommend meticulous management strategies, especially post-breakthrough, to uphold glove-protective performance. Conclusions Thermal aeration at 100°C for 1 hour proves effective, restoring protection without compromising glove strength. The study, covering twenty cycles, suggests safe glove reuse with proper decontamination, reducing costs significantly. However, limitations in chemical-glove combinations and exclusive focus on specific gloves caution against broad generalization. The absence of regulatory directives on glove reuse highlight the importance of informed selection and rigorous decontamination validation for workplace safety practices.

Objective To explore the role of apoptosis of neutrophils and vascular endothelial cells and changes of relative cytokines and thrombotic factors in the pathogenesis of traumatic sepsis at the advanced stage.Methods The venous blood was collected from the patients with traumatic sepsis at advanced stage and traumatic patients without sepsis and healthy subjects.The peripheral hlood mononu- clear cells(PBMC)and neutrophils were isolated and cultured.The apoptosis of neutrophils and vascular endothelial cells was assayed,and the level of IL-4,IL-10 in PBMC culture supernatants were deter- mined,and the tissue factor(TF)and vW factor(vWF)of peripheral plasma were measured.Results The percentages of apoptosis of neutrophils and numbers of apoptotic circulating vascular endothelial cells were higher than that of traumatic patients without sepsis and healthy controls significantly.And the levels of anti-inflammatory cytokines IL-4,IL-10 and thrombotic factors TF,vWF in traumatic sepsis were elevated than that of traumatic patients without sepsis and controls too.Conclusion Immune suppres- sion and abnormal thrombotic state may be one characteristic of traumatic sepsis at advanced stage,which perhaps involveds in the pathogenesis of traumatic sepsis at the advanced stage and multiple organ dys- function syndrome.

OBJECTIVETo investigate the effect and its mechanism of salmon milt DNA (SMD) on age-related involutions in mouse thymus.

METHODSFemale BALB/C mice aged 10 months were divided randomly into three groups according to their weights. They were high dosage group (333.33 mg.kg(-1).b.w.d(-1)), low dosage group (166.67 mg.kg(-1).b.w.d(-1)) and control group (0 mg.kg(-1).b.w.d(-1)). After five weeks, their thymus indexes were measured and the thymocytes were counted and the thymus cortex thicknesses were also measured using Image-Pro Plus (version. 4.0) software in the thymus section. All the data were analyzed by SAS statistic software. Microarray technique was applied to screen the gene fragments, which were differentially expressed between the high dosage group and the control group, together with RT-PCR to further confirm some of them.

RESULTSNo significant differences of the variables including body weight, thymus weight and thymus indexes among the three groups were found. The thymocytes quantities of thymus cortex and medulla in the high dosage group were significantly higher than those of the control group (P < 0.01 and P < 0.05, respectively). The thymus cortex thicknesses of both SMD supplement groups were significantly higher than that of the control group (P < 0.01 and P < 0.05 respectively). 112 differently expressed gene fragments were isolated. Furthermore, we found the fragments with the logged number of U23789, X80232 and Aw209102 were highly expressed in the high dosage group when RT-PCR technique was used.

CONCLUSIONSSMD may reverse the age-related involutions in mouse thymus via up-regulation the expression of proliferation related genes and via up-regulation the expression of development and differentiation related genes simultaneously.

Actins ; genetics ; Aging ; drug effects ; genetics ; Animals ; Cell Division ; drug effects ; genetics ; DNA ; administration & dosage ; Female ; Gene Expression ; drug effects ; Mice ; Mice, Inbred BALB C ; Pilot Projects ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Salmon ; genetics ; Thymus Gland ; cytology ; drug effects ; metabolism

The Formosan sika deer (Cervus nippon taiouanus) is an endemic subspecies in Taiwan. The original wild deer has been extinct since the late 1960s. The largest captive population is located at the Taipei Zoo. Except for infectious disease outbreaks, no systemic medical research has been reported for this subspecies. This study was conducted to analyze the medical status of the captive Formosan sika deer population, including the hematological and serum chemistry characteristics. To accomplish this, medical records for 34 Formosan sika deer from January 2003 to January 2014 were acquired and analyzed. The most common illness and cause of death was trauma, followed by gastrointestinal and respiratory disease, respectively. The hematologic and serum chemical values of healthy adults were quite different from those of sika deer (Cervus nippon yesoensis). This study provides a closer medical understanding of this subspecies and the results will facilitate its management.
Adult ; Cause of Death ; Chemistry ; Deer* ; Disease Outbreaks ; Gastrointestinal Diseases ; Humans ; Medical Records ; Retrospective Studies* ; Taiwan