Objective:To develop a fixed clamp apparatus for Macrophage system and PLEKHQ1 gene knockout mice, in order to provide a safe, simple, convenient and time-saving experimental tool for mice tail blood collection and injection.Methods:Clamp for macrophage system and PLEKHQ1 gene knockout mice was made by 50 ml syringe with slot according to the drawing. Clamp base was made by 50 ml plastic tube and appropriate size of the plastic tap.Results:Compared with current mice clamp in the market, the mice fixed clamp was a simple, low-cost, safe, easy and timesaving tool.Conclusion: The mice clamp for mice tail blood collection and injection provided a safe and convenient tool, and greatly improved the efficiency of the experiment.

Objective To explore the treatment opportunity of mechanical ventilation for acute severe organophosphorus pesticide poisoning patients with respiratory failure.Methods According to the different indications of mechanical ventilation,115 acute severe organophosphorus pesticide poisoning patients with respiratory failure were divided into the observation group A (40 cases),the observation group B(38 cases)and the control group(37 cases).The cure rate,aspiration pneumonia incidence,duration of mechanical ventilation,length of hospital stay of the three groups were analyzed.Results The cure rate of the observation group A(97.5％) was significantly higher than that of the control group(83.4％)and the observation group B (92.1％) (P ＜ 0.05 or P ＜ 0.01) ;The aspiration pneumonia of the observation group(2 cases) was significantly less than that of the control group(8 cases)(P ＜ 0.05) ;The hospitalization time [(4.46 ± 0.51) d,(7.11 ± 0.83) d] and the duration of mechanical ventilation [(55.40 ± 5.24)h,(79.47 ± 2.44) h] of the observation group A and the observation group B were significantly shorter than those of the control group [(10.40 ± 1.12) d,(100.30 ± 2.15) h] (all P ＜ 0.01) ; The hospitalization time and the duration of mechanical ventilation of the observation group A [(4.46 ± 0.51) d,(55.40 ± 5.24) h] were significantly shorter than those of the observation group B [(7.11 ± 0.83)d,(79.47 ± 2.44)h] (all P ＜ 0.01).Conclusion Respiratory rate ≥30 times/min and ＜ 34 times/min or ≤ 10 times/min,or spontaneous breathing weakened,and blood gas analysis prompted hypoxemia(60 mm Hg ＜ PaO2 ≤ 70 mm Hg)is best opportunity of mechanical ventilation for acute severe organophosphorus pesticide poisoning patients with respiratory failure.

OBJECTIVE:To observe the clinical efficacy and safety of ambroxol hydrochloride in the adjuvant treatment of el-derly patients with chronic obstructive pulmonary disease. METHODS:94 elderly patients with chronic obstructive pulmonary dis-ease were randomly divided into control group(48 cases)and observation group(46 cases). Control group was given oxygen inha-lation,antispasmodic asthma,expectorant and antitussive,antibiotics against infections,bronchodilation drugs,glucocorticoids and other conventional treatment;observation group was additionally given 30 mg Ambroxol hydrochloride injection,adding into 200 ml 5% glucose injection by intravenous infusion,twice a day. The treatment course for both groups was 2 weeks. The clinical efficacy,improvement of cough and sputum volume before and after treatment in 2 groups were observed,and incidence of ad-verse reactions was recorded. RESULTS:The clinical efficacy in observation group was significantly higher than control group,re-ducing degree of cough and sputum volume were significantly superior to control group,the differences were statistically significant (P<0.05);and there were no significant differences in the incidence of total adverse reactions,nausea,stomach heartburn,rash and indigestion between 2 groups(P>0.05). CONCLUSIONS:Based on the conventional treatment,ambroxol hydrochloride can effectively relieve symptoms and improve efficacy in the adjuvant treatment of elderly patients with chronic obstructive pulmonary disease,with no obvious adverse reactions and reliable safety.

Objective To study the pharmacokinetics of aristolochic acid A in Radix Aristolochiae and the compound preparation of Guanxinsuhe Capsule in mice in vivo after single-dose oral administration and observe the difference of aristolochic acid A absorption and distribution. Methods Aristolochic acid A assay was performed by RP-HPLC on a Waters apparatus with a DiamonsilTM C18 column (250 mm × 4.6mm, 5 μm), a mobil phase: a mixture of methanol-water-acetic acid (72: 27 : 1), flow rate: 1.0 mL/min, detection wavelength: 315 nm, and column temperature: 20 ℃. Results Mice were given Radix Aristolochiae and Guanxinsuhe Capsule by ig at the same level of 2. 5 mg/kg of aristolochic acid A, respectively, which were suspended in 0. 3% CMC-Na solution. Plasma concentrations were determined by RPHPLC. After single-dose ig administration of Radix Aristolochiae or Guanxinsuhe Capsule to mice, the mean plasma concentration-time courses of aristolochic acid A obtained fitted the one-compartment model.The main pharmacokinetic parameters of aristolochic acid A in Radix Aristolochiae, t1/2ka, t1/2 ke, tmax,AUC, Cmax are 5. 103 min, 43. 63 min, 17.89 min, 80. 45 (μg · min)/mL, and 0. 916 8 μg/mL; the rela tive pharmacokinetic parameters in Guanxinsuhe Capsule are 5. 294 min, 43.50 min, 18. 32 min, 33.08(μg · min)/mL, and 0. 381 8 μg/mL. Conclusion The Cmax of aristolochic acid A in Guanxinsuhe Capsule is significantly less than that in Radix Aristolochiae, which indicates that the compound compability could decrease the absorption of aristolochiae acid A.

Objective To investigate the roles of hemopoietic stem cells (HSCs) in the pathogenesis of psoriasis. Methods HSCs were isolated from the bone marrow of a patient with psoriasis vulgaris and a normal human control. Forward- and reverse-subtracted cDNA libraries were constructed by using suppression subtractive hybridization (SSH) technique between HSCs from the patient and control. Bacterial PCR and dot hybridization were performed to screen for positive clones followed by gene sequencing, identification and functional analysis. Real time quantitative PCR was carried out to measure the mRNA expression of interferon-γ and thymosin 10 (TMSB10). Results Nine genes were screened from the forward-subtracted cDNA library which encoded interferon-γ, tyrosine phosphatase, SUMO1 activase, etc, and 8 genes including the TMSB10-encoding gene were screened from the reverse-subtracted cDNA library. The relative expression level of interferon-γ in HSCs from the patient was 47.5 times that in HSCs from the control, while the level of TMSB10 from the control was 22.6 times that from the patient. Conclusion The abnormal expression of 17 genes which encode interferon-γ, thyrosin, and so on, may be involved in the development of psoriasis.

BACKGROUND: Alginic sodium diester (ASD) possesses neuroprotective function because of its selective calcium antagonist effects.OBJECTIVE: To compare the influences of ASD on intraneuronal Ca2+content and nerve cell apoptosis before and after reperfusion in focal cerebral ischemic rats.DESIGN: Randomized controlled observation.SETTING: Neurological Department of Xiangya Hospital Affiliated to South China University; Laser Orthopedic Surgery of the First Hospital Affiliated to Southern China University.MATERIALS: This experiment was carried out between November 2003and April 2004 at the Neurological Department of Xiangya Hospital Affiliated to South China University. A total of 65 male SD rats were recruited and randomized into 6 groups; 17 got lost during the experiment, and the other 48 rats completed the experiment with 8 rats in each group.METHODS: In sham operation group, an incision was made on rats' cervical skin and sutured. Right cerebral middle artery was occluded in rats of ischemic group, ASD 5 mg/kg preischemic group, ASD 5 mg/kg postischemic group, ASD 10 mg/kg preischemic group, and ASD 10 mg/kg postischemic group. After that, rats in all but ischemic group were subjected to intraperitoneal injection of various dosage of ASD or excipient 30minutes before reperfusion and 5 hours after reperfusion. FCM was used to determine intraneuronal Ca2+ content and rate of nerve cell apoptosis;meanwhile, neurological dysfunction was scored.MAIN OUTCOME MEASURES: [1] Influence of ASD on the score for neurological dysfunction, intraneuronal Ca2+ fluorescence intensity, and neuronal apoptosis in rats with right cerebral middle artery ischemia. [2]Correlation of behavioral obstacle score with intraneuronal Ca2+ fluorescence intensity and neuronal apoptosis in rats with right cerebral middle artery ischemia.RESULTS: Totally 65 rats were enrolled in this study, 17 of which got lost and the other 48 rats entered the result analysis. [1] Influence of ASD on the score for neurological dysfunction, intraneuronal Ca2+ fluorescence intensity, and neuronal apoptosis in rats with right cerebral middle artery ischemia: The score was obviously reduced in ASD 5 mg/kg preischemic group, ASD 5 mg/kg postischemic group, ASD 10 mg/kg preischemic group and ASD 10 mg/kg postischemic group as compared with ischemic group (1.80±0.21, 2.20±0.23, 1.20±0.11, 2.00±0.22, 3.40±0.65); moreover,functional improvement was more obvious due to pre-reperfusional administration than post-reperfusional administration. Intraneuronal Ca2+ concentration was reduced after ASD administration at different degrees and lower than that of ischemic group. Decrement of intraneuronal Ca2+ concentration was found most obvious due to 10 mg/kg ASD administration 30 minutes before reperfusion, approximately reduced by 70%; moreover, neuronal apoptosis rate on the ischemic side was obviously suppressed by ASD administration, displaying time-dependent manner, with apoptotic suppression effect more obvious in pre-reperfusional group than in post-reperfusional group (5.68%, 10.03%; 4.00%, 9.91%). [2] Correlation of behavioral obstacle score of right cerebral middle artery ischemic rats with intraneuronal Ca2+ fluorescence intensity and membrane associated protein/propidium iodide apoptosis: Obvious positive correlation was found between behavioral obstacle score and intraneuronal Ca2+ fluorescence intensity and detection rate of membrane associated protein/propidium iodide apoptosis (r=0.51,0.62, P ＜ 0.05); intraneuronal Ca2+ fluorescence intensity was also positively correlated with the detection rate of membrane associated protein/propidium iodide apoptosis (r=0.84, P ＜ 0.05).CONCLUSION: [1] ASD can exert anti-apoptosis effect by suppressing the increment of intraneuronal Ca2+ concentration, thus having neuroprotective function and ultimately improving neurological dysfunction. [2] Its effect displays time-dependent manner, and neurological functional improvement is more obvious by pre-reperfusional administration than by post-operational administration.

Objective To evaluate the use safety of a unitary package peracetic acid ( PAA) disinfectant through acute and subacute toxicity experiments in rats.Methods This study was conducted according to the Technical Standard For disinfection ( Version 2002 ) .In the acute toxicity experiment, 60 healthy Wistar rats were randomly divided into different groups of PAA given by single intragastric gavage.Symptoms of poisoning of the animals were observed to calculate the half lethal dose ( LD50 ) .In the subacute toxicity experiment, 40 healthy Wistar rats were randomly divided into three dose groups and negative control group.The rats were given 33-342 mg/kg BW PAA for 28 days.At the end of test, the body weight, the organ-to-body weight ratio and the hematological and biochemical indices were determined, and pathological examination was performed.Results The acute oral LD50 of female rats was 1470 mg/kg BW, and the LD50 of male rats was 1710 mg/kg bw.The results of subacute toxicity test showed that the body weight, the organ-to-body weight ratio and the hematological and biochemical indices had no significant difference between the dose groups and control group.No obvious abnormality was observed in gross anatomy and histopathological examination.Conclusions The results of the acute toxicity test prove that PAA has a low toxicity level, and no significant subacute toxicity was observed at the exposure doses.

Objective To conduct an empirical study for quantifying the anastomosis between two vessels after skin expanded technique by the method of angiography and to provide a precise basis for vascular study in skin flap.Methods Bilateral skin flaps based on deep iliac circumflex vessels were elevated from the abdominal wall including deep superior epigastric vessels.One was expanded at the boundary between two vessels and the other unexpanded.An X-ray image was obtained by carotid arterial injection of gelatin-lead oxide mixture.Three parallel lines with equal intervals perpendicular to long axis of the two vessels were designed at the communication area.Vessel anastomosis quantity was determined by counting the number of marks derived from the intersections of the lines and the vessels and statistical analysis was carried out.Results The marks of intersection in expanded group were more than unexpanded group with statistical significance.Conclusions The method for quantifying vessel anastomosis in skin flap is reliable.The principles of this procedure may also be applied to other experimental and elinical studies.

Through looking up Chinese Pharmacopoeia and searching for unannounced inspection on the website of China food and drug administration and some GMP regulations and gudelines of European Union, the problems existing in the raw materials control of animal original biochemical drugs were discussed, and the relevant suggestions were put forward.