Objective To simply the method of how to obtain cultures rich in astrocytes from SD rat cerebral cortex which could be utilized in vitro experiments.Methods The neonatal rat cerebral cortex was made into suspension by mechanical dissociation,and then reduced other cells by differential velocity adherent technique,shaking in orbital shaker and passage of cultured cells.After purification,the cultured cells were identified by double immunofluorescence staining and SABC.Results We successfully obtain cultures rich in astrocytes and the proportion of astrocytes were more than 95%.Conclusion The method described above was reliable in obtaining the high purity astrocytes from neonatal rat cerebral cortex and double immunofluorescence staining was more vivid and direct.

As the advanced technology of proteomics, the surface-enhanced laser desorption/ionization Time-of-Flight Mass Spectrometry(SELDI-TOF-MS)has great potential in clinical application for it′s technological features.Whether it can be applied in clinical trials depends on the quality control and standardization of the technology.In the article, we introduced and discussed the following aspects of the issue.1) quality controlling before sample being analyzed, including the collection and preparation of samples, the selection and conservation of reagents, the calibration and attendance of laboratory apparatus;2) quality controlling during the assay, that is, the validation and analysis of the experimental result;3) the standardization of the continued data handling and analyzing, including the standardization of the analyzed data′s nature, the selection and standardization of the data′s analyzing methods, the system evaluation of the whole SELDI-TOF-MS technological system.

Objective To investigate the effect of progesterone pretreatment of focal cerebral ischemic and reperfusion injury (fCIRI) and underlying molecular mechanisms.Methods A single intraperitoneal injection of progesterone (8 mg/kg) given 1 h,48 h and 96 h before fCIRI was established in male Sprague-Dawley rats.The number of survival of neurons in hippocampal CA1 region of the ischemiaside,as well as spatial memory function,was detected on days 3-8 after fCIRI.Extracellular-signalregulated kinase 1/2 phosphorylation (p-ERK1/2) and nuclear translocation of p-ERK1/2 in hippocampal CA1 region were examined using western blot.Results The number of survival of neuronal cells was significantly increased in ischemic groups treated with progesterone at 1 h and 48 h pre-fCIRI (164.3 ± 11.0,218.5 ± 9.1 and 142.7 ± 12.1,F =29.4,P ＜ 0.01) compared with fCIRI group treated with vehicle.Likewise,the escape-latency to reach the hidden-platform recorded in day 5 of Morris water maze test was reduced markedly in fCIRI-treatment groups compared with the vehicle group(10.3 ± 11.1,19.2 ±9.6 and 32.4 ± 14.3 ;F =35.8,P ＜0.01).The level of p-ERK1/2 was elevated notably during 24 h to 48 h postprogesterone by western blot,while restored to the baseline at 96 h post-progesterone.Improved nuclear translocation of p-ERK1/2 was observed from 2 h to 48 h post-progesterone.The progesterone receptor antagonist RU486 blocked the exaltation of either intracellular level or nuclear translocation of p-ERK1/2,which was induced by progesterone.Conclusions The pretreatment with progesterone exerts a neuroprotective effect against the ischemia-induced neuronal death and ameliorates the deficits in spatial memory through enhancing the activation of ERK1/2.The neuroprotection derived from pretreatment with progesterone achieves a time window of not less than 48 h,which is progesterone receptor-mediated ERK1/2 signaling pathway-dependent.

AIM To set up the model of stom- effects of its treatment drugs. METHODS To ap-ach smooth muscle cells contraction and assess the ply the culture of isolated stomach fundus smooth muscle cells in rats in vitro and to obtain the mixture of stomach smooth muscle cells. The cells were examined with the Trypan blue dilution technology under microscope and the length of cells and the contraction rate of muscle cells were adopted to the evaluation index of effects. The change of cells in different time was observed and calculating the survival rate and time of cells. The selected neuro-medium and its antagonist, such as acetyl-choline chloride (ACh) and atropine, 5-HT and domperidone(Domp) , histamine( His) and diphen-hydramine(Diph) were added to the mixture cells so as to provoke irritant reaction observe the effects of drugs. RESULTS Obtaining cells in stomach fundus smooth muscle was more than 1 X 106 in each rat by this way, and the survival rate and time of cells was 95% and 2. 5 h. The contraction of cells was observed after adding ACh 5-HT or His ,and the effects of these agents were antag- onized by antagonist atropine, Domp and Diph respectively. The effects of maximum contraction of ACh in concentration 1?10-8 mol ? L-1 was 38%, and in 5-HT may exist two receptors, and the maximum rate was 20. 7% and 10. 5% respectively. The effective dose of contraction was 1 X 10-13 -1?10-10 mol ? L-1 induced by His, the contraction effects was significantly weakened in high of 1 ?10-10 mol ? L-1 and antagonized by Diph. CONCLUSION The smooth muscle cells by digesting stomach fundus smooth muscle of rats would be more than 95% survivors and survival time was more than 2. 5 h. ACh,5-HT and His could induce the contraction of smooth muscle cells, and the effects of contraction were antagonized by atropine, Domp and Diph respectively.

OBJECTIVE To investigate the value of acute phase protein in the diagnosis and therapy for nosocomial infection monitoring. METHODS The levels of serum C-reactive protein(CRP),serum amyloid A(SAA),?_1-acid glycoprotein(AAG),and ?_1-antitrypsin(AAT) were measured in 71 gynecological patients with nosocomial infection during chemotherapy,Thirty normal controls and 33 chemotherapy patient controls were detected by nephelometry and compared with white blood cell counts. RESULTS The nosocomial infection rate of chemotherapy patients was 16.7%.Main pathogens were Gram-negative bacteria(60.6%).Fungi infection rate was 19.7%.The most frequent hospital infection sites were respiratory tract,gastrointestinal tract and wound.Compared with controls,the levels of serum CRP,SAA,AAG and AAT were significantly higher in chemotherapy patients with bacteria infection(P

Objective:To investigate the KRAS gene mutation features in lung and gastric cancers and their relationship with clinicopathologic characteristics. Methods:A total of 128 lung cancer and 115 gastric cancer patients were included. Polymerase chain reaction amplification and DNA sequencing were conducted to detect mutations in exon 2 of the KRAS gene. Results:The mutation frequency of KRAS was different in lung and gastric cancers;however, it did not show any statistical significance (6.3%vs. 4.3%, P>0.05). The KRAS codon 12 gene mutation ranks the first in both types of cancer. No significant correlation was observed between the prevalence of KRAS mutations and patient's age and gender. KRAS gene mutation rate was higher in lung adenocarcinoma than in non-adenocarcinoma, such as squamous cancer (10.7%vs. 0%, P<0.05). Conclusion:No correlation was found between the KRAS gene mutation and the sex and age of lung and gastric cancer patients in Jiangsu Province. The rate of KRAS mutation was low. KRAS gene mutation rate was relatively higher in lung adenocarcinoma patients;thus, the mutation status of the KRAS gene should be evaluated be-fore undergoing EGFR-TKI therapy.

Objective To study the association of tryptophan hydroxylase-2 (TPH2) gene polymorphism and antisocial personality disorder (ASPD) and its impulsivity in Chinese Han population. Methods The single nucleotide polymorphism (SNPs) of TPH2 in transcriptional control region,-703G/T,was analyzed by PCR-RFLP genotyping assay in 117 ASPD patients and 142 healthy controls. Barratt Impulsiveness Scale-11 (BIS-11) was used to evaluate the impulsivity of subjects. Results There were significant differences between ASPD and controis on genotype and allele frequencies of TPH2-703G/T (x2 = 7.73, P ＜ 0.05; x2 = 5.12, P ＜ 0.05). The GG genotype and G allele were positively associated with ASPD(OR = 1.458,95% CI = 1.080 ～ 1.968 ;OR = 1.479,95% CI = 1.045 ～ 2.094). The scores of BIS-11 and its factors in GG genotype group((71.28 ± 7.50), (19.60 ±3.41), (25.73 ± 4.92), (25.95 ± 4.77) ) were higher than GT genotype group (( 66.23 ± 8.06), (17.79 ±3.02) ,(23.06 ±3.84) ,(25.38 ±4.97)) and TT genotype group((66.55 ±8.49),(18.50 ±3.35),(23.45 ±4.08), (24.97 ± 4.90)), but only the difference of BIS-11 total scores, the attention and motor factor scores among three groups were statistically significant (P＜0.05). The scores of BIS-11 and its factors in G allele group ((69.38 ±8.04), (18.92 ± 3.36), (24.73 ±4. 69), (25.73 ±4.82)) were higher than T genotype group ((66.41 ±8.22),(17.98 ±3.26),(23.27 ±3.94), (25.15 ±4.89)),however,only the difference of BIS-11 total scores, the attention and motor factor scores between two groups were statistically significant.Conclusion TPH2-703G/T polymorphism may be association with ASPD in Chinese Han population. The GG genotype and G allele may be the risk factors of ASPD and impulsivity.

Objective To investigate the effect of prevention and treatment of urinary retention for patients after abdominal surgery using electroacupuncture unit model 6805.Methods 180 cases of ab-dominal surgery patients were randomly divided into three groups, the treatment group(64 patients)were treated with electroacupuneture at the points of sanyinjiao, zhongji, pangguangyu, the control group 1(57patients)Was treated with moxibustion and foment were used in the control group 2(259 patients).The incidence rate of urinary retention Was compared between the three groups. Results The rate of spontaneously emiction was 56.25%in 30 minutes and 35.94%in 40 minutes in the treatment group, the results in the control group 1 and 2 were 22.81% and 15.79%,23.73% and 13.56% respectively. There were significant differences between three groups. Conclusions Electroacupuncture Can effectively prevent and treat urinary retention after abdominal operation.

ObjectiveTo study the effect and mechanisms of nourishing Piyin Remedy (nPR) and bovine brain extract (bBE) on experimental spinal cord injury (SCI) of rat.Methods80 healthy SD rats were divided into 5 equal groups randomly: bBE group supplied through subarachnoid cavity, normal saline (NS) group supplied through subarachnoid cavity, nPR group, NS orally taken group, combined group. Animal models were made by Allen's equipment on T8～T9 segment. The spinal nerve function, somatosensory evoked potentials (SEP), retrograde and label technique of horseradish peroxidase, gross observation, histological and morphometric analysis were taken as the observed indices.ResultsThe values of observed indices of bBE group and nPR group improved evidently compared with their own control groups; that of combined group was prior to sole administration.ConclusionnPR can hold back the secondary SCI and accelerate the recovery of spinal nerve function; bBE can stimulate the improvement of injuried nerve fibers; the joint of nPR and bBE can make a synergic effect.

ObjectiveTo study the therapeutic effect and mechanisms of nourishing Piyin Remedy (nPR) on experimental rat spinal cord injury.MethodsThirty-two healthy SD rats were divided into 2 equal groups randomly: nPR group and injured group. Animal model of incomplete injury of spinal cord was made by Allen's equipment on rat's T8—T9 segment. The spinal nerve function,SEP,retrograde and label technique of horse radish peroxidase,gross observation,histological and morphometric analysis were taken as the observed indices.ResultsThe value of observed indices of nPR group were improved evidently compared with injured group.ConclusionNourishing Piyin Remede can hold back the secondary spinal cord injury and accelerate the recovery of spinal nerve function.