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.

Objective To evaluate the efficacy of integrated traditional Chinese medicine(TCM)and western medicine in treatment of acute exacerbations of chronic obstructive pulmonary disease(AECOPD)combined with respiratory failure,TCM syndromes of spleen-kidney-yang deficiency and phlegm-dampness by comparison between the integrated therapy and simple western therapy in treatment of the disease. Methods 160 patients with AECOPD combined with respiratory failure,spleen-kidney-yang deficiency and phlegm-dampness syndrome in the intensive care units(ICU)of Affiliated Hospital of Chengdu University of TCM and other four hospitals were randomly allocated into two groups in this double-blinded,multicenter,prospective,randomized,controlled trial. In the control group (78 cases),western medicine and placebo were given to the patients,and in the treatment group(82 cases), conventional western medicine plus fei-shuai mistura 25 mL were administered,four times per day,the therapeutic course lasting for 2 weeks in both groups. The all-cause mortality,respiratory failure-cause mortality,improvement of modified Medical Research Council(mMRC)Dyspnea Scale grades,6 minutes walk distance(6MWD),the forced expiratory volume in 1 second/forced vital capacity(FEV1/FVC)were observed in the 28 days after the end of treatment. Results In the comparisons between the control and treatment groups,there were no statistical significant differences in the all-cause mortality〔54.87%(45/82)vs. 64.10%(50/78)〕and the cases of FEV1/FVC(both P>0.05)in the 28 days after the end of treatment;the 28 day respiratory failure-cause mortality was significantly decreased〔19.51%(16/82)vs. 33.33%(26/78),P<0.05〕,the number of patients with mMRC Dyspnea Scale grades (1-2)was obviously increased(22 cases vs. 7 cases,P<0.05),and the number of patients with 6 MWD grades (4-6)was markedly enhanced in the treatment group(21 cases vs. 8 cases,P<0.05). Conclusions The integrated TCM and western medicine has better therapeutic results in improvement of the patients' degree of dyspnea, 6 MWD and respiratory failure mortality than simple treatment with western therapy for treatment of patients with AECOPD combined with respiratory failure, spleen-kidney-yang deficiency and phlegm-dampness syndrome. However,in regard to the effect on pulmonary function and all cause mortality,the integrated therapy for treatment of such patients in short term has no significant effect.

AIM: To observe the effects of six common traditional Chinese herbs of activating blood, paeoniae rubra radix, salviae miltiorrhizae radix, ligustici, rhizome, notoginseng radix, pruni persicae semen and wine staemed radix et rhizome, on atherosclerotic plaque structure and stabilization in ApoE gene-deficient mice. METHODS: Four sections of the aortic root were choosen and stained with hematoxylin and masson. All sections were measured with Image-ProDR○ Plus Version 4.5.1 (IPP) software. RESULTS: Compared with the model group, plaque area corrected by cross-sectional vessel wall area reduced significantly in salviae miltirrhizae radix treatment group, lipid core area reduced in paeoniae rubra radix group, pruni persicae semen and wine steamed radix et rhizome treatment group, minimum thickness of fibrous cap became thicker significantly in salviae miltiorrhizae radix, ligustici, rhizome, pruni persicae semen and wine steamed radix et rhizome treatment group. CONCLUSION: These Chinese herbs may stabilize the atherosclerotic plaques in ApoE gene-deficient mice by interfering their structure, but their effects do not parallel with their activating blood efficacy in traditional Chinese medicine.

The cupping spot is considered as one kind of skin change due to cupping treatment. With literature regarding cupping spot, the influencing factors and value of cupping spot in clinical diagnosis and treatment were analyzed, which could make a further exploration on the action mechanism of cupping treatment. The literature showed that the formation of cupping spot was related with cupping temperature, pressure, cup-retaining time, cupping area, individual difference and health condition, etc; cupping spot had the ability to assist diagnosis, prevent disease, cure disease and evaluate clinical efficacy. Previous studies on cupping spot have already made some progress, and played a positive significance on finding cupping rule and studying its mechanism. However, the research for this area is still in the primary stage, which needed deeper study to reveal scientific connotations of cupping spot.
Acupuncture Therapy ; instrumentation ; Humans ; Skin ; anatomy & histology ; blood supply

OBJECTIVETo study the possible relationship between genetic polymorphism of N-acetyltransferase 2 (NAT2) and susceptibility to hepatocellular carcinoma.

METHODSGenetic polymorphisms of the four NAT2 genes in 78 patients with hepatocellular carcinoma and 112 healthy controls were analyzed by means of real-time fluorescence light-Cycler. The difference in frequencies between the hepatocellular carcinoma patients and the controls were compared.

RESULTSThe significant difference in slow acetylation genotype frequency was found between the controls and the hepatocellular carcinoma patients who were smokers (17.9% vs 37.5%, x(2)= 4.67, P<0.05) resulting in increased by 2.76 times the risk for hepatocellular carcinoma, but no evident difference between the controls and hepatocellular carcinoma patients who were non-smokers.

CONCLUSIONThe smokers with slow acetylation genotype of N-acetyltransferase 2 may be the population with high risk for hepatocellular carcinoma.

Acetylation ; Adult ; Aged ; Arylamine N-Acetyltransferase ; genetics ; Carcinoma, Hepatocellular ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Liver Neoplasms ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Smoking ; genetics

Objective To establish an automatic synthesis method for 11C-carfentanil (CFN) as an novel opiate receptor radioligand and study its biodistribution in rats. Methods 11C-Triflate-CH3 was bubbled into 0.5 mg precursor desmethyl-CFN (which was dissolved in 0.15 ml DMSO) to generate 11C-CFN in a V-tube at room temperature. Sep-Pak C2 column was used for purification of 11C-CFN, which was eluted by 3ml binary system aqueous solution, 10 ml water thrice, and then I ml ethanol. The biodistribution (% ID/g) of 11C-CFN in SD rats was studied. SPSS 13.0 was used for statistical analysis. Non-normal distribution data were analyzed using nonparametric test. Results The synthesis time for 11C-CFN was 20 min (end of bombardment, EOB). The synthesis yield was (35.5 ± 2.2) % on average (n = 12, uncorrected)with the radiochemical purity over 98%. Biodistribution study in rats showed that the tracer had a high brain uptake, rapid blood clearance, and a metabolic pathway via liver and kidney. The highest tracer uptake was in thalamus (4.26 ± 0.89) % ID/g and striatum (4.05 ± 1.08) % ID/g at 5 min after injection, followed by cerebral cortex (2.63±0.89) %ID/g, pons (2.26 ±0.57) % ID/g, hippocampus (2. 17 ±0.55) %ID/g and cerebellum (2. 15 ±0.39) %ID/g. Conclusions The automatic synthesis of 11C-CFN is fast and reliable, and this radioligand can be used for opiate receptor imaging.

OBJECTIVETo compare the clinical therapeutic effects differences between acupuncture at Suliao (GV 25) and Shuigou (GV 26) on promoting regain of consciousness from coma in severe craniocerebral injury.

METHODSBased on regular emergency treatments of neurosurgery, eighty-two cases of craniocerebral injury who were under stable condition were randomly divided into an observation group (42 cases) and a control group (40 cases). Suliao (GV 25) was selected as main aupoint, while Laogong (PC 8) and Yongquan (KI 1), etc. were selected as adjuvant acupoints and Neiguan (PC 6), Sanyinjiao (SP 6), Yifeng (TE 17) and Wangu (GB 12), etc. were selected as matching acupoints in the observation group where a strong needle manipulation was applied to improve the regain of consciousness. The main acupoint of Shuigou (GV 26) along with identical adjuvant acupoints and matching acupoints in the observation group were selected in the control group with identical strong needle manipulation. The treatment was given once a day in both groups, five times per week and ten times were considered as one session. The immediate clinical symptoms after acupuncture at Suliao (GV 25) and Shuigou (GV 26) were observed as well as Glasgow coma scale (GCS) before the treatment, after 45 days and 90 days of treatment to assess the resuscitation time and rate. Also the clinical efficacy was compared between both groups.

RESULTSThe occurrence rate of sneezing reflex was 85.7% (36/42) in the observation group, which was higher than 25.0% (10/40) in the control group (P < 0.01). The average resuscitation time was (64.6 +/- 19.4) days in the observation group, which was obviously shorter than (73.8 +/- 14. 6) days in the control group (P < 0.05). The resuscitation rate was 88.1% (37/42) in the observation group, which was similar to 75.0% (30/40) in the control group (P > 0.05). Compared before the treatment, GCS were both improved after the treatment in two groups (both P < 0.01). The 90-day GCS was 9.52 +/- 2.32 in the observation group, which was superior to 8.47 +/-2.14 in the control group (P < 0.05). The curative and markedly effective rate was 45.2% (19/42) in the observation group, which was superior to 22.5% (9/40) in the control group (P < 0.05).

CONCLUSIONThe effect of acupuncture at Suliao (GV 25) on improving regain of consciousness from coma in severe craniocerebral injury is positive. It could specifically improve sneezing reflex and stimulate respiratory center, which has more obvious effect than acupuncture at Shuigou (GV 26).

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Coma ; etiology ; physiopathology ; psychology ; therapy ; Consciousness ; Craniocerebral Trauma ; complications ; Female ; Humans ; Male ; Middle Aged ; Young Adult

Objective To evaluate the value of diffusion tensor imaging(DTI)in cognitive impairment of patients with acute cerebral infarction.Methods Diffusion tensor images were obtained from 30 volunteers who underwent clinical MR imaging and were found to have no abnormalities on conventional MR images and 30 patients who were clinically diagnosed cerebral infarction and were found to have infarction lesions on conventional MR images.Color-coded FA images and three-dimensional color-coded tensor images were reconstructed.For volunteers,average apparent diffusion coefficient(ADC)and fractional anisotropy(FA)were measured in some main white matter structures of peripheral white matter, basal ganglia,and cerebral peduncle,etc.For infarction patients,ADC and FA were measured and compared between infarction lesions and corresponding contralateral normal regions.Pearson correlation analysis was used to determine correlation with cognitive impairment.Results In infarction patients group, FA and ADC of lesions unrecovered declined.Change in ADC and FA had positive correlation with cognitive impairment of patients with acute cerebral infarction.Conclusion DTI has positive correlation with cognitive impairment of patients with acute cerebral infarction.

OBJECTIVETo study the microanatomy of the perforating arteries in the superior space of the internal carotid artery visualized through a pterional approach.

METHODSTwelve (24 sides) perfused cadaver heads were dissected via the pterional approach, and the perforating arteries in the superior space of the internal carotid artery were studied under microscope. The diameter, course and distribution in the anterior perforated substance of the perforating arteries were recorded.

RESULTSAll the perforating arteries exposed lied on the side of the brain tissue. The carotid bifurcation on 8 sides had perforating arteries, and 11 sides showed medial lenticulostriate artery of the middle cerebral arteries, with short course and overlapped with another perforating arteries upon entry into the anterior perforated substance. On 4 sides, the medial lenticulostriate artery coincided with the perforating arteries in A1. All 24 sides showed middle lenticulostriate artery and lateral lenticulostriate artery of the middle cerebral arteries. Most of the lenticulostriate arteries originated from the anterior segment of the bifurcation of the middle cerebral arteries. The earlier bifurcation occurred in M1 of the middle cerebral arteries, the more perforating arteries originated. On 22 sides, the anterior cerebral arteries had perforating arteries with many branches, and fewer perforating arteries in a main artery were associated with greater diameter of them.

CONCLUSIONThe superior space of the internal carotid artery allows a space for operation, and in some cases, part of the medial leticulostriate arteries and A1 perforating arteries can be severed to obtain larger space for the operation.

Brain ; anatomy & histology ; blood supply ; surgery ; Cadaver ; Carotid Artery, Internal ; anatomy & histology ; surgery ; Female ; Humans ; Male ; Microsurgery ; Neuroanatomy ; methods

OBJECTIVETo compare the clinical, pathological, laboratory test and follow-up data between familial and sporadic patients with distal myopathy with rimmed vacuoles (DMRV) and discuss the characteristics of this disorder in Chinese population.

METHODSThe clinical and pathological features, laboratory data and follow-up results of 33 sporadic and 4 familial cases of pathologically confirmed DMRV were summarized and compared retrospectively.

RESULTSThe patients age, onset age, or disease duration showed no significant difference between sporadic and familial cases; the onset pattern and affected muscle groups were also similar, but the sporadic cases showed more frequent dysmorphic features than the familial cases. The patients showed mild to moderate elevation of the muscle enzymes by one to three folds, and the familial patients had more significant elevation than the sporadic ones. No correlation was found between the disease duration and the level of muscle enzymes. The pathological findings were similar between the cases, and Gomori staining showed rimmed vacuoles and inclusion bodies without inflammatory cell infiltration. Follow-up results of 29 cases showed no significant difference between the two groups. The disease was slowly progressive and severely affected the quality of life of the patients, but did not produce obvious effect on the life expectancy.

CONCLUSIONThe clinical, pathological and laboratory data of Chinese DMRV patients are basically similar to those of Japanese cases. Sporadic cases tend to show more dysmorphic features than the familial ones, and occasional sporadic cases have early disease onset in early childhood.

Adult ; Asian Continental Ancestry Group ; Distal Myopathies ; classification ; genetics ; pathology ; Female ; Humans ; Inclusion Bodies ; pathology ; Male ; Pedigree ; Retrospective Studies ; Vacuoles ; pathology ; Young Adult