Objective To observe the expression of 5-HT receptor 14 subtype mRNAs in the rat retina. Methods RT-PCR technique was used. Specific oligonucleotide primers were synthesized based on the complementary DNA sequence for each of the 14 cloned rat 5-HT receptor subtypes. Expression of 5-HT 1A、5-HT 2A、5-HT 2C、5-HT 3 and 5-HT 7 receptor subtype mRNAs were detected. Results Positive band densities of various 5-HT receptor subtypes were different in retina. The most bands were formed by 5-HT 7 and 5-HT 2A receptor subtypes, the secondary prominent bands were formed by 5-HT 2C and 5-HT 3 receptor subtypes. PCR products from each positively-detected receptor subtype were subcloned into pCRⅡ vector for sequencing and found in correspond to published complementary DNA sequences. No positive bands existed of 5-HT 1B, 5-HT 1D, 5-HT 1E, 5-HT 1F, 5-HT 2B, 5-HT 4, 5-HT 5A, 5-HT 5Band 5-HT 6receptor subtypes. Conclusions There may exist varieties of 5-HT receptor in rat retina, including 5-HT 1A, 5-HT 2A, 5-HT 2C, 5-HT 3 and 5-HT 7 receptor subtypes. It means the retina serotoninergic system may be a common feature in mammals and, by the light of the 5-HT receptor subtypes, play an important role in mediating the transmission of rod sensitive signals.

Immunocytochemical staining technique by using specific antibody against 5-HT1A receptor subtype (5-HT1AR) wasused to observe the distribution of 5-HT1AR immunoreactivity in the rat nervous system. The highest level of 5-HT1AR im-munoreactivity was observed in piriform cortex, septum, ventraldorsal thalamic nucleus, reticular thalamic nucleus, basolateralamygdaloid nucleus, Purkinje cell layer, red nucleus, facial nucleus and nucleus of the trapezoid body. Considerably weaker im-munoreactivity was detected in hippocampus, frontal cortex, mediodorsal thalamic nucleus, interpeduncular nucleus, mesen-cephalic trigeminal nucleus, dorsal raphe nucleus, spinal trigeminal nucleus, the superficial layers of the spinal dorsal horn, dor-sal root and trigeminal nerve ganglia, Very weak immunoreactivity was found in the olfactory bulb, caudate putamen,globus pal-lidus, nucleus diagonal band, bed nucleus stria terminalis, habenular nucleus, substantia nigra and superior olive. The presentresults indicate that 5-HT1AR immunoreactive structures are widely distributed in the rat nervous system and might play impor-tant role in mediating the multiple effects of 5-HT in the nervous system.

Aim To investigate the expression of vascular endothelial growth factor(VEGF) and its receptors in human hepatocellular carcinoma cell lines SMMC7721, HHCC and HepG2. Methods Immunohistochemical staining and reverse-transcriptase polymerase chain reaction(RT-PCR) were used to detect the mRNA and expression of VEGF and its receptors: VEGF-R1(Flt-1) and VEGF-R2 (KDR) in three human hepatocellular carcinoma cell lines, SMMC7721, HHCC and HepG2, as compared with ECV304 cells(human umbilical vein endothelial cells) and L929 cells(mouse fibroblast). Results All three human hepatocellular carcinoma cell lines expressed VEGF protein. Flt-1 mRNA and protein could be detected in SMMC7721 cells while KDR in HHCC and HepG2 cells. Conclusion The expressions of Flt-1 and KDR suggests that VEGF may be an autocrine growth factor for human hepatocellular carcinoma, at least for cell lines in vitro.

The present study was designed to examine the morphological pattern of primary afferent projections into the spinal dorsal horn by labeling the lumbar dorsal root ganglia with carbocyanine fluorescent dye DiI in mouse embryos and neonatal pups aged embryonic day 12 to postnatal day 3 (E12-P3). Primary afferent fibers projected into dorsal funiculus at E13, but did not penetrated into gray matter of dorsal horn until E15. The afferent projections became dense and entered the spinal gray matter more deeply at E16 and E17. By E18 the intensity of primary afferent in the deep part of the dorsal horn increased and their branching patterns became more complicated. Some of these primary fibers were also observed to ramify extensively in the superficial laminae. The projection pattern of primary afferent remained unchanged after birth, but the intensity of afferent terminals increased in the superficial laminae. In addition, afferent fiber collaterals that projected into the contralateral dorsal horn were also observed. They were first examined at E16 and mainly originated from the medial and deep part of the dorsal horn. Around birth, the contralateral projections were also found to originate from the lateral part of dorsal horn. Our results indicate that laminar organization of primary afferents in the spinal dorsal horn is established during the late embryonic and early postnatal stages.This organization then undergoes further refinement to match the pattern seen in the adult.

AIM To study the inhibitory effect of the cytotoxin (CTX) from guangxi cobra venom on human nasopharyngeal cancer cell (CNE) and other tumons cells. METHODS The cytotoxin was isolated and purified from Guangxi cobra venom by successive chromatography on Sephadex G-50 and CM Sepharose CL-6B columns. Its cytotoxic effects and does-effect relationship on human tumors cell lines were examined by MTT assay. RESULTS The inhibitory effects of CTX CM-5 on CNE, human ovarian carcinoma cell (Ho8990), uterus cervical carcinoma cell (HELA) and lymphoma (YAC) cell lines showed a definite does-effect relationship. The IC 50 (48 h in cubation) was 1.84, 2.59, 1.84 and 0.75 mg?L -1 respctively. The inhibitory effects of CTX CM-5 on CNE increased with time. The IC 50 (3 h and 24 h cubation) was 4.78 and 1.04 mg?L -1 respctively. CONCLUSION CTX from Guangxi cobra venom exhibites strong suppressive effect on cultured tumor cells line in vitro.

Objective\ To observe the expression of 5\|HT 1A and 5\|HT 2A receptor mRNAs in the rat spinal cord and dorsal root ganglion(DRG). Methods\ In situ hybridization histochemical technique. Results\ (1)5\|HT\-\{1A\} receptor mRNA positive neurons were found in all laminae of the gray matter,mainly in the superficial laminae(laminae Ⅰ and Ⅱ),laminae Ⅲ and Ⅳ of the dorsal horn.Scattered positive neurons were also observed in laminae Ⅴ\|Ⅶ and Ⅹ.Very few positive signals were found in the ventral horn(lamnina Ⅸ);(2)5\|HT\-\{2A\} receptor mRNA positive neurons were mainly found in the superficial laminae and ventral horn,while sparsely distributed positive neurons were also located in other laminae.Within the DRG:(1)About 10\^4% of the total DRG cells were labeled with 5\|HT 1A receptor mRNA.The positive signals were mainly confined to a subpopulation of small\| and medium\|sized cells;(2)About 17\^4% of the total DRG cells were labeled with 5\|HT 2A receptor mRNA,most of them were also small\| and medium\|sized cells. Conclusions\ 5\|HT 1A and 5\|HT 2A receptor mRNAs positive neurons distributed heterogeneously in the rat spinal cord and DRG, they may play important roles for 5\|HT in the analgesic effects at the spinal level and nociception in the periphery.\;

BACKGROUND: Long-term sustained release naltrexone has been reported in clinical application near one year that it can improve emotional state and relieve addiction; therefore, the effect of long-term sustained release naltrexone on memory restoration at neuropsychology level were explored. OBJECTIVE: To observe the effect of long-term sustained release naltrexone on memory ability of opioid addicts. DESIGN, TIME AND SETTING: A contrast observational study was performed at Drug Rehabilitation Centers of Wuhan, Changde, Zhengzhou, and Jiangyang between October and December 2006. Healthy controls were tested in Shenzhen Hospital of Peking University in October 2006. PARTICIPANTS: A total of 88 males with opioid addicts were divided into naltrexone group (n=35), compulsory detoxification group (n=26), and non-treatment group (n=27). Another 22 healthy subjects were considered as the controls. METHODS: At 6-12 months before testing, naltrexone (3.1 g) was subcutaneously implanted into bilateral abdominal wall in the naltrexone group; patients in the compulsory detoxification group underwent completely compulsory detoxification for 6 months, and the examination results, including diamorphine, methadone, and buprenorphine in urine, were negative on the immediately testing day. Event related potential and its wave form were recorded from the opioid addicts in the three groups and from healthy controls who finished semantic recognition between new and old words using portable-type event related potential working system. MAIN OUTCOME MEASURES: Correct rate and response time of semantic memory; latency and amplitude of language related potential-N400. RESULTS: ①There were significant differences in correct rate and response time between three expedmental groups and healthy control group (P ＜ 0.001 ); in addition, correct rate was significantly increased, and response time was significantly shortened in the naltrexone group compared to compulsory detoxification group and non-treatment group (P＜ 0.05). ② N400 latency in the three experimental groups was significantly longer than healthy control group (P ＜ 0.01), while N400 amplitude in the naltrexone group was increased, which was no significant difference compared to healthy control group (P> 0.05) but was significant difference compared to compulsory detoxification group and non-treatment group (P ＜ 0.01). CONCLUSION: Long-term sustained release naltrexone can effectively improve neural function and enhance semantic memory of the opioid addicts.

objective To detect CK19 mRNA and CEA mRNA in blood of the peripheral vein and to investigate the effect of ligating pulmonary vein firstly or ligating pulmonary artery firstly during surgical operation on haematogenous dissemination of malignant cells.Methods Fifty six non-small cell lung cancer patients were collected and random assigned to two groups before operation (ligating pulmonary vein firstly or ligating pulmonary artery firstly).The patients were accepted radical operation and their operations were put in practice by doctors of one team.Vein blood was collected one day before operation and one week after operation.CK19 mRNA and CEA mRNA in blood were detected by nested reverse transcriptase-polymerase chain reaction. Result The positive rate of expression of CK19 mRNA and CEA mRNA after operation is lower than that before operation.The positive rate of expression of CK19 mRNA and CEA mRNA in ligating pulmonary vein firstly after operation is lower than that in the other group. Conclusions Surgical operation have effects on the dissemination of malignant cell and ligating palmonary vein firstly during operation can reduce the dissemination of malignant cell.

OBJECTIVE: To explore the protective effects of ischemic postconditioning on non-heart-beating donor (NHBD) in rat NHBD lung transplantation model. METHODS: Forty Sprague-Dawley rats were randomized into the ischemic postconditioning group (IPO group) and the control group (C group), 10 pairs in each group in which left lung orthotopic transplantations from NHBDs were done with " two-cuff-one-stent technique". In the C group, perfusion was resumed by declamping pulmonary artery immediately after transplantation, whereas in the IPO group, 5 cycles of 1-min reperfusion and 1-min reocclusion of pulmonary artery were applied as postcontioning before full recovery of perfusion. RESULTS: Compared with the C group, water content of donor lungs was lower and pathological changes were milder in the IPO group, meanwhile compliance, structure and function of donor lungs were better preserved. Furthermore, the expression of cell apoptosis and MDA content in donor lungs were lower in the IPO group, while SOD content was higher. CONCLUSION Ischemic postconditioning can reduce ischemic reperfusion injury of NHBD lung transplantation and preserve the structure and function of donor lungs. It can inhibit lipid peroxidation and cell apoptosis in NHBD lungs after transplantation.
Animals ; Heart Arrest ; Ischemic Postconditioning ; Lung ; metabolism ; pathology ; surgery ; Lung Transplantation ; Male ; Models, Animal ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Superoxide Dismutase ; metabolism

Objective:To explore the effects of dopamine receptor D2(DRD2)on astrocytic dedifferentiation based on SOX2-regulated genes in neural stem cells(NSCs)and astrocytes.Methods:Immunofluorescence staining and SOX2-GFP mice were used to examine the lineage differentiation of SOX2-positive cells during the development of cere-bral cortex.Primary NSCs/astrocytes culture,ChIP-seq and Western Blot were adopted to analyze and verify the expres-sion of candidate genes.Pharmacological manipulation,neurosphere formation,photochemical ischemia,immunofluo-rescence staining and behavior tests were adopted to evaluate the effects of activating DRD2 signaling on astrocytic dedif-ferentiation.Results:Immunofluorescence staining demonstrated the NSC-astrocyte switch of SOX2-expression in the normal development of cerebral cortex.ChIP-seq revealed enrichment of DRD2 signaling by SOX2-bound enhancers in NSCs and SOX2-bound promoters in astrocytes.Western Blot and immunofluorescence staining verified the expression of DRD2 in NSCs and reactive astrocytes.Application of quinagolide hydrocholoride(QH),an agonist of DRD2,signifi-cantly promoted astrocytic dedifferentiation both in vitro and in vivo following ischemia.In addition,quinagolide hydro-choloride treatment improved locomotion recovery.Conclusion:Activating DRD2 signaling facilitates astrocytic dedif-ferentiation and may be used to treat ischemic stroke.