Background Channelrhodopsin-2 (ChR2)is a cation channel isolated from the eyespot of Chlamydomonas algae and has been used to control neuron activity.The light stimulation is a more precise fashion whether space or time than that of electrical,magnetic and ultrasound stimulation. Objective This study was to construct a replication deficient recombinant adenovirus cxpression vector of ChR2 and to determine its function.Methods Human embryo kidney 293 (HEK293) cell line was cultured and passaged in DF12 medium containing 10％ fetal bovine serum(FBS).The ChR2 gene was cloned at the downstream of cytomegalovirus(CMV)promoter of the adenoviral shuttle plasmid pSB291 in sense direction,and the resultant recombinant plasmid pSB291-hChR2- GFP was transfected into HEK293 cell together with plasmid pBHG lox ( deltaE1,3 ) containing adenoviral genome,then small amounts replication deficient recombinant adenovirus expression vector of ChR2 (Ad-ChR2) was obtained.Through amplification gradient centrifugation and dialysis,pure Ad-ChR2 was obtained.Visual cortex cells derived from 4 1-day-old clean Long Evans rats were primary cultured with serum-free culture media and infected by AdChR2.When expressing green fluorescencc,those cells received the stimulated of blue light with 460 nm.Patch clamp technique was applied to record an action potential. Results After purification and concentration,the titer of AdhCHR2 reached 7.9×1010 PFU/ml.Twenty-four hours after transfect of Ad-ChR2,HEK293 cell membrane showed the green fluorescence for the recombinant plasmid with green fluorescence protein under the inversed fluorescence microscope.The HEK293 cells change their shape from flat to round 13 days after transfected.The primary cultured visual cortex cells exhibited the green fluorescence 3-5 days after infected by Ad-ChR2.The action potentials evoked by blue light stimulation were recorded with patch clamp on those cells expressing green fluorescence. Conclusions Ad-ChR2 expressing vector is constructed successfully in this study.It is verified that Ad-ChR2 expressing vector can infect visual cortex cells with visual function.This result is very important for visual plasticity study.

This study was aimed to explore the mechanism of Jia-Yan-Kang-Tai (JYKT) on Th1/Th2 immune cell imbalance in rats with autoimmune thyroiditis (AIT).A total of 40 Lewis rats were made into AIT rat model.Six week later,AIT rats were randomly divided into the model group,low-dose,middle-dose and high-dose JYKT group,with 10 rats in each group.Intragastric administration of deionized water,0.708 g· kg-1,1.417 g· kg-1 and 2.834 g· kg-1 JYKT was given to rats in each group,respectively for 8 consecutive weeks.Serum TgAb and TPOAb were detected.The ratio of Th1 cell,Th2 cell and Th1/Th2 was measured.Contents of IL-4,IL-5,IFN-γ,TNF-α and IL-1oα in plasma were detected.The pathology of thyroid tissues was observed by HE dyeing.The protein expressions of IFN-γ and IL-4 in thyroid tissues were observed under immunohistochemical staining.The results showed that high-dose JYKT group can reduce the level of TPOAb and TGAb,the incidence of thyroiditis,the ratio of Th1 cell,Th2 cell and Th1/Th2;decrease contents of IFN-γ,TNF-α,IL-1α,IL-4 and IL-5 in plasma;reduce the protein expression of IFN-γ and IL-4 in thyroid tissues.It was concluded that JYKT was able to suppress thyroid autoantibodies of AIT rats,which may be related to the regulation of cytokine level and Th 1/Th2 immune cell imbalance.

This study aimed to investigate the effects of kaempferol on the skeletal muscle of KKAy mice via PI3K-AKT-GLUT4 signaling.Spontaneous type 2 diabetic KKAy mice were made up for the model group.After 6-week treatment of kaempferol by intragastric administration,key targets of PI3K-AKT-GLUT4 signaling were detected using biochemical and immunohistochemical technique,and western blot.It was found that the body weight,fast blood glucose and random blood glucose were decreased after kaempferol administration.ITT results show that blood glucose at 30 min after injecting insulin and the area under the curve drop were reduced in the kaempferol group,and so was the insulin resistance index in the kaempferol group.In addition,the mRNA expressions of PI3K,AKT and GLUT4 in the kaempferol group increased significantly,while the protein levels of AKT and GLUT4 were up-regulated by kaempferol administration.It was concluded that kaempferol can significantly regulate the blood glucose,body weight and insulin resistance in mice through activating the PI3K-AKT-GLUT4 signaling.

This study aimed to elucidate the mechanism behind the effects of oleuropein (OL) on improving insulin resistance in obese db/db mice.Twelve 6-to 8-week-old male db/db mice were randomly divided into the model group and the OL group with 6 in each group according to their levels of blood glucose and body weight,while six C57BL/6J mice with the same age were made up for the normal group.Mice of the OL group was intragastrically administered with (50 mg·kg-1) once a day.The mice of the other groups were treated with the saline solution with the same dosage.After treatment for four weeks,OGTT test was carried out,while fast blood glucose and serum insulin were tested.RT-PCT and western blot were used to quantify the mRNA and protein expressions of certain targets in the liver.As a result,it was found that the body weight,fast blood glucose,serum insulin level and insulin resistance index were significantly decreased in the mice of the OL group when compared with model group after the treatment for 4 weeks (P＜0.05 or P＜0.01).Plasma glucose levels at each time point of OGTT tests were lower in the mice of the OL group than those of the model group (P＜0.01).The mRNA and protein expressions of InsR,IRS-1 and GLUT-2 in the mice of the OL group increased significantly (P＜0.01).In conclusion,it was demonstrated that oleuropein may reduce the blood glucose and improve the insulin resistance in db/db mice through up-regulating the mRNA and protein expressions of InsR,IRS-1 and GLUT-2 in the liver.

AIM To compare the percutaneous penetration performance of three paeonol gels.METHODS Franz diffusion cell method was applied to investigating the penetration and retention behaviors of eutectic mixturebased nanoemulsion,ordinary nanoemulsion and saturated solution gels onto mouse skins in vitro.The retention and permeation amounts in stratum corneum and hair follicles of volunteers smeared with gels were compared by tape stripping method.RESULTS The accumulative permeation and retention amounts of various gels onto mouse skins in vitro were in sequence of eutectic mixture-based nanoemulsion gel ＞ ordinary nanoemulsion gel ＞ saturated solution gel.The main retention of all the three gels was observed on the volunteers' skin surface,and the permeation amounts of eutectic mixture-based nanoemulsion,ordinary nanoemulsion gels,and their accumulative permeation amounts in stratum corneum and hair follicles were significantly higher than those of saturated solution gel (P ＜0.05).CONCLUSION Nanoemulsion technology can significantly promote the percutaneous penetration performance of paeonol.

This study aimed to explore the impacts of Cyclocarya paliurus polysaccharide (CP) on insulin signal pathway in liver cells.H4IIE liver cells of rat that cultivated for 72 h were made up for the model group.H4IIE cells at 70％-80％ confluence were exposed to various concentrations of CP,including 50,100,200 and 400 μg·mL 1,for measuring cell viability using Neutral Red.Based on the results of cell viability,H4IIE cells were divided into the control group,the insulin group (100 nmol·L-1),the low dose CP group (the LCP group,200 μg· mL-1) and the high dose CP group (the HCP group,400 μg· mL-1).After the cultivation of cells for 2 h,mRNA levels were measured by real-time RT-PCR,while phosphorylation levels of target proteins were detected by western blot after the treatment for 30 min.It was found that the cell viability indexes in the cells administered by 100,200 and 400 μg·mL-1 CP increased significantly compared with the control group (P＜0.01).After the administration for 2 h,InsR and IRS-2 mRNA expressions in the insulin group,the LCP group and the HCP group increased (P＜0.05 or P＜0.01).After the administration for 30 min,phosphorylation levels of InsRβ,IRS-2 and Akt in the insulin group were higher than those in the HCP group (P＜0.01).In conclusion,CP probably increased the cell viability of H4IIE cells,and improved the blood glucose index through enhancing the mRNA expressions of InsR and IRS-2 and the phosphorylation levels of InsRβ,IRS-2 and Akt in the liver.

OBJECTIVETo observe the efficacy differences of acupoints massage for asthenopia of video display terminal (VDT) under different exposure dose.

METHODSOne hundred and two cases (204 eyes) were divided into a low exposure group and a high exposure group, fifty-one cases in each group. The same intervention of acupoints massage on Cuanzhu (BL 2), Jingming (BL 1), Sizhukong (TE 23), Sibai (ST 2) and Taiyang(EX-HN 5) were given to the two groups, one acupoint for 5 min and once everyday, one month of which made a course. The symptom score, tear film break-up time (BUT) and Schirmer I test(SIT) were observed before and after treatment.

RESULTS(1) The correlation coefficient of cubic curve model of the exposure dose was the biggest with symptom improvement index (P = 0.000), which indicated that the lower VDT exposure index was, the more obvious the symptom improved. The symptom improvement indices of low exposure group and high exposure group, which were (52.31 +/- 16.65)% and (28.93 +/- 13.35)% respectively, were statistical significant difference (P = 0.000). (2) Compared to before treatment, the levels of BUT and SIT in the two groups were both significantly higher (P < 0.05). Compared with the high exposure group, the levels of BUT and SIT in the low exposure group were increased by 0.826 s (P = 0.022) and 1.029 mm (P = 0.033), respectively, after the impact of BUT and SIT was corrected before the research.

CONCLUSIONThe acupoints massage can improve the symptoms and ocular physiology for patients with VDT asthenpia, and it is more effective for the low exposure cases.

Acupuncture Points ; Adult ; Asthenopia ; physiopathology ; therapy ; Computer Terminals ; utilization ; Female ; Humans ; Male ; Massage ; Middle Aged ; Occupational Diseases ; physiopathology ; therapy ; Tears ; secretion ; Treatment Outcome ; Young Adult

Objective For the second children diagnosed as ABO hemolytic disease of the newborn(ABO-HDN),we made a comprehensive analysis of the related indicators of prenatal and postpartum,so as to achieve early prevention,early diagnosis and early treatment.Methods Prenatal microcolumn gel assay was used to detect the father and mother's blood type and mother's irregular antibody,mother serum IgG anti A(B)anti-body titer.Microcolumn gel assay was used to detect hemolysis three tests after the production of pregnant women.The results were divided into five groups according to the results of hemolysis three tests:group A[di-rect antiglobulin test(+),free antibodies test(+)and antibody releasing test(+)],group B[direct antiglobu-lin test(-),free antibodies test(+)and antibody releasing test(+)],group C[direct antiglobulin test(+), free antibodies test(-)and antibody releasing test(+)],group D[direct antiglobulin test(-),free antibod-ies test(-)and antibody releasing test(+)]and group E[direct antiglobulin test(+),free antibodies test (-)and antibody releasing test(-)].Total bilirubin,unbound bilirubin,hemoglobin,reticulocyte percentage and lactate dehydrogenase were detected by automatic analyzer.Results ABO-HDN children hemolysis three tests,in the 5 groups,the higher the titer of the mother's IgG anti A(B)antibody,the more serious the child' s condition was,the difference was statistically significant(P<0.05).Reticulocyte percentage and lactate de-hydrogenase in the five groups,the difference was statistically significant(P<0.05).Conclusion A combina-tion of antenatal and postnatal multiple laboratory test parameters is more accurate in predicting the second child ABO-HDN.At the same time,it helps to master the state of the disease and reduce the occurrence of complications and sequelae.

Objective To investigate the microsurgical anatomy of the jugular foramen and its adjacent structures so as to provide the anatomical base for the surgical approach and nerve preservation in the region. Methods With the aid of the surgical microscope, a combined approach was performed,which included transjugular approach, extreme lateral trans-condylar approach and infralabyrinthine approach. The jugular foramen and its adjacent structures were exposed from multi-directions step by step to identify the spatial relationships of important structures in this region. Results At the inner wall of skull, dural septations divided the jugular foramen into the petrosal portion, the sigmoid portion and the intrajugular compartment. The dura overlying the intrajugular compartment had two meatus, both of which were medial to the intrajugular processes. One was the glossopharyngeal meatus, and the other was the vagal meatus. The structures that traversed the jugular foramen were the sigmoid sinus, inferior petrosal sinus, jugular bulb, glossopharyngeus nervus, pneumogastric nerve, accessorius, meningeal branches of the ascending pharyngeal and occipital arteries, and the cochlear aqueduct. Conclusions It has more surgical significance to describe the petrosal, sigmoid, and intrajugular portions of the jugular foramen. The asterion can be defined as the landmark of the junction of transverse sinus and sigmoid sinus. Detailed micro-anatomic study may improve the success of surgery, protect cranial nerves and prevent unnecessary injury.

Hedyotis hedyotidea has been traditionally used for the treatment of arthritis, cold, cough, gastro-enteritis, headstroke, etc. But few studies have screened the active compounds from extracts of H. hedyotidea. In this study, the structure of the chemical constituents from stems of H. hedyotidea were determined and the immunosuppressive activity of the compounds was evaluated. The compounds were separated and purified with silica gel, gel column chromatographies and preparative HPLC, and their structures were identified by spectral methods such as MS and NMR. Eleven compounds were obtained and identified as(6S,9S) -vomifoliol (1), betulonic acid (2), betulinic acid (3), betulin(4), 3-epi-betulinic acid (5), ursolic acid (6), β-sitosterol (7), stigmast-4-en-3-one (8), 7β-hydroxysitosterol (9), (3β,7β) -7-methoxystigmast-5-en-3-ol (10) and morindacin (11). This is the first report of compounds 1, 2, 4, 8, 9, 10 and 11 from H. hedyotidea. Compounds 1, 2 and 8-11 were firstly isolated from the genus Hedyotis, and compounds 9 and 10 were isolated from the family Rubiaceae for the first time. The immunosuppressive activity of these compounds was tested using the lymphocyte transsormationtest. Compounds 4, 6 and 9 showed significant immunosuppressive activity.
Animals ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Hedyotis ; chemistry ; Immunosuppressive Agents ; chemistry ; isolation & purification ; pharmacology ; Lymphocytes ; drug effects ; immunology ; Male ; Mass Spectrometry ; Mice ; Mice, Inbred C57BL ; Molecular Structure ; Plant Stems ; chemistry