Dopamine(DA) modulates diverse wake-related behaviors including movement,reward, and cognition.Dopaminergic neurons are located in the substantia nigra pars compacta and ventral tegmental area.There are five distinct DA receptors(R):D_1R,D_2R(D_(2S)R and D_(2L)R), D_3R,D_4R and D_5R in the central nervous system, in which D_1R and D_2R are majorly expressed. The affinity of D_2R for endogenous DA is significantly higher than that of D_1R.Re- cently,studies by pharmacological and gene knock-out animals revealed that dopamine D_2R is essential inmaintaining wakefulness.Here,we review the progress on roles of D_2R in sleep-wake regulation.

Ring finger protein 43 (RNF43) is a ring-type E3 ubiquitin ligase.As a negative regulater of Wnt signaling pathway, RNF43 has an important anti-tumor effect.The mutation of RNF43 may cause abnormal activation of Wnt signaling pathway, and then promote invasion, metastasis and proliferation of tumor cell.In addition, the act of RNF43 protein in the Wnt signal pathway is expected to be a molecular target in the therapy of cancer.In recent years, with the gradual deepening of related research, the molecular structure of RNF43 protein and its mechanism of action with the Wnt pathway-related proteins have been gradually clear.In clinical, RNF43 protein analogs and related vaccine also show the important position in the therapy of cancer.

Objective To detect the expression and function of ZNRF3 in different kinds of thyroid cancer tissues and cells.Methods Immunohistochemistry was used to detect the expressions of ZNRF3 protein in 35 cases of papillary thyroid carcinoma and 10 cases of poorly differentiated thyroid carcinoma.The expressions of ZNRF3 gene in TPC-1 and 8505C were detected by RT-PCR,and the cell lines were derived from papillary thyroid carcinoma and poorly differentiated thyroid carcinoma respectively.After silenced ZNRF3 gene expression with lentivirus,the proliferation ability of TPC-1 cells were detected with CCK-8,the invasion and metastasis ability of TPC-1 cells were detected with Transwell.Results According to results of immunohistochemistry and RT-PCR,the expressions of ZNRF3 in papillary thyroid carcinoma cells and tissues were higher than those in poorly differentiated thyroid carcinoma cells and tissues,the differences were statistically significant (4.83±0.44 vs.3.13 ±0.59,t =2.20,P ＜0.05;1.01±0.06 vs.0.21±0.04,t =11.80,P＜0.01).After ZNRF3 geng silencing,according to the results of CCK8,the proliferation ability of TPC-1 cells was significantly enhanced in 72 h,the difference was statistically significant (0.96 ± 0.10 vs.0.64 ± 0.05,t =3.19,P ＜ 0.05);and according to the results of Transwell,the TPC-1 cell's invasion (0.12 ± 0.01 vs.0.09 ±0.00,t =5.48,P＜0.01) and migration (0.22 ±-0.01 vs.0.17 ±0.01,t =4.58,P ＜0.05) also increased,the differences were statistically significant.Conclusion The expression of ZNRF3 in papillary thyroid carcinoma is higher than that in poorly differentiated thyroid cancer.ZNRF3 is tumor suppressor gene in the thyroid tumors.

Recurrent laryngeal nerve injury is one of the serious complications after thyroidectomy.Unilateral injury causes hoarseness,while bilateral injury causes difficulty in breathing or even life-threatening glottis obstruction.Analyzing the root cause of the thyroid injury,firstly it is the anatomical factors of recurrent laryngeal nerve,namely the close and complex relationship between the recurrent laryngeal nerve and the inferior thyroid artery,the existence of branches of recurrent laryngeal nerve and its variation,and the presence of non recurrent laryngeal nerve and Zuckerkandl nodules.Those are all made the recurrent laryngeal nerve easy to be damaged.Secondly it is because of the vulnerability of the recurrent laryngeal nerve itself.Last improper using of energy operation instrument will cause heat injury on nerves.Below counter measures can be implemented to prevent recurrent laryngeal nerve injury.Dissect to show recurrent laryngeal nerve or make it ‘ visualization’ during thyroidectomy.Elaborately anatomize recurrent laryngeal nerve to appropriate degree.Be familiar with the property of energy operation instrument and thus safely use them to reduce the heat injury of recurrent laryngeal nerve.Reasonably use the intraoperative nerve monitoring in the surgery,which assist to reduce the risk of injury of recurrent laryngeal nerve.

Objective To investigate the heat effects of the high-frequency electric knife on the recurrent laryngeal nerve ( RLN) in pigs and the safety margin in which electric knife can be used .Methods Totally 12 pigs for experiment were randomly divided into 3 groups by the distance between the head of the electric knife and the nerve(2 mm, 1 mm, 0 mm).The application time of the electric knife touching RLN was set to be 3 s and the application energy of the electric knife was 90 W.The data of electromyogram were measured by means of nerve detector before and after operation .Statistical analysis was made based on those data .The nerve tissues were taken to make paraffin sections so that the histological change can be observed and compared before and after damage.Results The data from electromyogram by nerve detector indicated that the difference of the amplitude of group 2(1 mm)and group 3(0 mm)had statistical significance(P<0.05)within group.There were significant difference of the amplitude between the 3 groups.Histological study showed that the tissues of group 3(0 mm)had obvious injury .Conclusions In thyroid surgery , the safety range of high frequency electrical knife used around RLN is:the distance from electrical knife head to nerve should be no less than 2 mm.

Objective To investigate the effect of heat stress on organ indices, intestinal morphology, gastric mu-cosal HSP70 mRNA expression and glucose metabolic hormones in mice.Methods A single-factor experiment was de-signed for the present research.Eighteen KM mice of the similar age and weight were randomly divided into control group and heat stress group.The weight of the heart, liver, spleen, lung and kidney, as well as the expression of HSP70 mRNA in the mouse gastric mucosa were measured.The plasma concentration of insulin and glucagon, the villus height and crypt depth of the duodenum and jejunum were detected.The histological changes of the liver, duodenum and jejunum were also examined.Results There was no effect of heat stress on the organ indices.It significantly increased the expression of HSP70 mRNA in the gastric mucosa, reduced the plasma insulin level and caused serious injury to the liver, duodenum and jejunum in the mice.Conclusions Heat stress does not significantly affect the organ indices in mice, but can significantly increase the expression of HSP70 mRNA in the gastric mucosa, cause apparent damages in the liver, duodenum and jeju-num, reduce the villus height, crypt depth and villus height/crypt depth ratio of the duodenum and jejunum, and also de-crease the blood insulin concentration in the mice.

Objective To study the preparation of hepatitis C viruses (HCV) genotyping oligochip and its application in the detection of 76 hepatitis C patients.Methods Oligonucleotide probes and primers were designed in the 5’noncoding region and core region of HCV. The HCV typing chip was prepared by spotting the modified probes onto nylon membrane. Products of the second PCR were labeled with Dig-dUTP. Furthermore, 6 PCR products were sequenced.Results Using the chip,15 subtypes in 11 types of HCV were analyzed.Results of hybridization indicates that 76 hepatitis C patients were all positive and 20 health people were negative.Among 76 patients, 64 cases were 1b type, 11 cases were 2a type and 1 case was 3a type. Mix infection was not found. The results obtained by sequencing 6 samples and chip arraying were the same.Conclusion The HCV genotyping chip could be used in detecting serum HCV RNA and analyzing its genotypes.

OBJECTIVETo isolate and elucidate the chemical constituents of the fruits of Acanthopanax sessiliflorus.

METHODIsolation and purification were carried out on the column chromatography of silica gel and Sephadex LH-20. Their structures were elucidated on basis of physicochemical properties and spectral data.

RESULTNine compounds were isolated and identified as oleanolic acid-3-O-6'-O-methyl-beta-D-glucuronopyranoside (1), 22-alpha-hydroxychiisanogenin (2), oleanolic acid-3-O-beta-D-glucuronopyranoside (3), oleanolic acid-3-O-beta-D-glucopyranoside (4), oleanolic acid (5), chiisanogenin (6), (-)-sesamin (7), daucosterol (8), beta-sitosterol (9).

CONCLUSIONCompound 1 is obtained from the genus Acanthopanax genus for the first time. Compounds 2-5 are isolated from this plant for the first time.

treating rheumatoid arthritis.　METHODS: Eighty patients with rheumatoid arthritis were randomly divided into two groups with each group having 40 patients. Group one (M 8, F 32; age 46 a± s 11 a; disease history 63 mo±48 mo) was treated with anti-inflammation sub-group No.1 and No.3. Group two (M 6, F 34; 44 a±9 a; disease history 45 mo±45 mo) was treated with sub-group No.2 and No.4. One week before the initiate of the study, the originally used non-steroid anti-inflammation drugs were stopped for all two-groups patients and each patient took 2 tablets of oxaprozin po qn. At the beginning of the study the patients received 2 tablets of anti-inflammation drugs No.1 daily and 6 tables of No.3 weekly in 1st group, and 2 tablets of anti-inflammation drugs No.2 daily and 6 tablets of No.4 weekly in 2nd group respectively.　RESULTS: In the leflunomide group, the total effect rate was 93 % and the remarkable improvement rate was 85 %. In the methotrexate group, the total effect rate was also 93 % and the remarkable effect rate was 83 %, P>0.05. Nine patients (23 %) in leflunomide group had adverse reaction as mainly skin itch, nettle-like rash, decrease of leukocytes, liver malfunction and others. Seventeen parients (43 %) in methotrexate group had adverse reaction as mainly responses of digestive tract, liver enzyme elevation, decrease of leukocytes, trichomadesis, manoxenia, and others.　CONCLUSION: Leflunomide has similar therapeutic efficacy to methotrexate. However, it has relatively less toxicity.

Objective To evaluate the effect of exosomes from different sources on the growth, metastasis and immune microenvironment of hepatocellular carcinoma ( HCC ) in a mouse model of orthotopic transplanted hepatoma so as to provide new insight into the clinical immunotherapy of hepatocellular carcinoma. Methods The serum-derived exosomes were obtained by ultracentrifugation from Hepa1-6 cells and 3-week orthotopically tumor-bearing HCC mice. The morphology and size of exosomes were identified by transmission electron microscopy. One-week tumor-bearing HCC C57BL/6 mice were randomly divided into 3 groups ( 3 mice for each group ) , and respectively treated with tail vein injection of 120 μl PBS(control group), 120 μl Hepa1-6 cell-derived exosomes (1μg/μl, TEXcel group), and serum-derived exosomes (1μg/μl) (TEXserum group). The treatments were conduced once a week for 3 consecutive weeks. The mice were sacrificed on the 4th day after the treatments, and the liver tissue and lung tissues were dissected. The volumes of the liver cancer tissues were measured. The expression of PD-L1 protein and CD3+and FoxP3+T lymphocytes infiltration in the liver cancer tissues were respectively detected by Western Blot and immunohistochemical staining. The lung metastasis of the liver cancer was detected by hematoxylin-eosin staining. Results The diameters of the Hepa1-6 cell- and serum-derived exosomes both were about 100 nm, and were uniform vesicles with complete membrane structure. Compared with the control group, Hepa1-6 cell-derived exosomes had a certain inhibitory effect on the growth and lung metastasis of HCC, while there was no significant difference for the TEXserum group. Western Blot results showed that compared with the control group, PD-L1 protein expression was significantly up-regulated in both TEXcel group and TEXserum group. Immunohistochemical staining showed that compared with the control group, the infiltration number of Foxp3+-labeled regulatory T cells (Treg) had no significant changes in TEXcel group and the TEXserum group, and the differences were no statistically significant (all P>0.05). However, the infiltration number of CD3+-labeled T lymphocytes was significantly reduced in the two groups, and the differences were statistically significant (all P<0.05). Conclusion The Hepa1-6 cell-derived exosomes have a certain inhibitory effect on the growth and lung metastasis of HCC, and have no obvious regulation effects on the immune microenvironment of HCC. The serum-derived exosomes from orthotopically tumor-bearing HCC mice can promote the growth and lung metastasis of HCC and immunosuppress the microenvironment. The Hepa1-6 cell-derived exosomes are expected to be used for immunotherapy studies of liver cancer.