The Telomeric Repeat Amplification Protocol(TRAP)and its modified versions change the size and/or the ratio of the telomerase products in the amplification stage of the assay.Based on TRAP,a useful method was developed for detecting telomerase activity in rice.A special precursor primer and a special reverse primer and conducted two steps of PCR cycles were designed.GENE Genius? Bio-imaging System was applied for this quantitative analysis for exploring telomerase activity and its optimal reaction conditions.The method ensured that the optimal reaction conditions for the telomerase was 19℃,13minutes,at a concentration of 0.28 u g/? l.A quantitative analysis method was established for detecting telomerase activity in rice.With this method,we detected telomerase activity in roots,young leaves and young panicles of six parental lines of hybrid rice.The results show that young panicles have the highest telomerase activity,demonstrating that telomerase activity is closely related to the cell vitality in plants.

Objective: To explore the mechanism of An-pressing manipulation in improving post-stroke muscle spasticity, by observing the changes of γ-aminobutyric acid (GABA) and glycine (Gly) in plasma and gray matter of L1-L3 spinal cord anterior horn in post-stroke rats with muscle spasticity after An-pressing manipulation intervention. Methods: Ten of 80 adult male Sprague-Dawley (SD) rats were randomly selected as the blank group, and the remaining 70 were used for modeling. The middle cerebral artery occlusion (MCAO) rat model was established by insertion suture occlusion method in the left external carotid artery. Thirty rats with a Longa neurological score of 2-3 points and a modified Ashworth spasticity scale score of 1-, 1+, or 2 were included in the experiment. Using the random number table method, the 30 successfully modeled rats were randomly divided into a model group, an An-pressing tendon group and an An-pressing muscle belly group. Two days after modeling, rats in the An-pressing tendon group and An-pressing muscle belly group received An-pressing manipulation on the tendon and belly of quadriceps femoris muscle respectively, with the pressure of (350±50) g and the frequency of 5 s/time, 15 min per session, once a day for 5 continuous days. After the 5th treatment, the tension of the rat quadriceps femoris muscle was evaluated using the modified Ashworth spasticity scale. The Gly levels in rat plasma and L1-L3 segments of spinal cord were determined by enzyme-linked immunosorbent assay (ELISA). The GABA levels in rat plasma and L1-L3 segments of spinal cord were measured by high performance liquid chromatography (HPLC). Results: The decrease in rat muscle tension scored by the modified Ashworth spasticity scale in the An-pressing tendon group was more significant than that in the An-pressing muscle belly group (P<0.01); the increases in Gly and GABA levels in the rat plasma and L1-L3 segments of spinal cord were more significant in the An-pressing tendon group than those in the An-pressing muscle belly group (all P<0.01). Conclusion: Based on the theory of 'anti-stretch reflex' of tendon organs, the use of An-pressing manipulation to induce the 'anti-stretch reflex' by stimulating the tendon organs can improve the muscle spasticity of rats, which is better than An-pressing the muscle belly. Increased levels of Gly and GABA in rat plasma and L1-L3 segments of spinalcord may be one mechanism of An-pressing manipulation to improve muscle spasticity by stimulating tendon organs.

Objective:To observe effect of acupuncture combined with hypothermia therapy on MAPK/ERK pathway and apoptosis related factorsin rats suffered cerebral ischemia reperfusion and to explore underlying mechanisms.Methods:Middle cerebral artery ischemia model were established.Ninety SD rats were randomly assigned into a blank group,a control group,a model group,an acupuncture group,a mild hypothermia group,and an acupuncture with hypothermia group.After 72 h treatment,nervefunction defect scores were observed,and infarction area percent was detected by 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) staining;expressions of Bcl-2 and Bax were examined by immunohistochemistry;apoptotic cells were detected by TUNEL assay;and expression levels of phospho-mitogen-activated protein kinase(p-MEK2) and phospho-extracellular signal regulated kinase 1/2 (p-ERK1/2) in the rats' hippocampus ischemic side were determined by Western blot.Results:In the rats of the model group,the neural function defect scores,the infarction area percent,the expression level of Bax,and apoptotic cells increased,while the level of Bcl-2 decreased significantly.The level of p-MEK2 and p-ERK1/2 increased obviously compared with the blank and control groups (P＜0.05 or P＜0.01).After treatment with acupuncture and hypothermia,the neural function defect scores,infarction area percent,and the level ofBax,apoptotic cells and the levels of p-MEK2 and p-ERK1/2 were significantly decreased,while the level of Bcl-2 in the treatment group was significantly elevated (P＜0.05 or P＜0.01) compared with the model group.Compared with the acupuncture group or the hypothermia group,the neural function defect scores and the levels of p-MEK2 and p-ERK1/2 in the acupuncture combined with hypothermia group were significantly reduced (P＜0.05 or P＜0.01).Conclusion:Acupuncture and hypothermia therapy can improve cerebral function,and reduce the cerebral injury through down-regulation of Bax level,and up-regulation of Bcl-2 level,which is related to reducing the levels of p-MEK2 and p-ERK1/2.The therapeutic effects on cerebral ischemia reperfusion injury for combination of acupuncture with hypothermia are better than those with single application of acupuncture or hypothermia.

Objective: To investigate the effect of acupuncture plus mild hypothermia on neurological function impairment score, cerebral infarct size and apoptosis-related factors in cerebral ischemia reperfusion injury (CIRI) rats.Methods: Sixty healthy male Sprague-Dawley (SD) rats were routinely reared for 1 week. Ten rats were randomly selected as the sham operation group and 10 rats as the blank control group, while the remaining 40 rats were subjected to preparing the middle cerebral artery occlusion (MCAO) model by modified filament occlusion method. The 40 MCAO rats were further randomly divided into a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, with 10 rats in each group. Rats in the sham operation group, the blank control group and the model group did not accept treatment except binding; rats in the acupuncture group received acupuncture treatment; rats in the mild hypothermia group received mild hypothermia treatment; rats in the acupuncture plus mild hypothermia group received acupuncture and mild hypothermia treatment. 72 h after the treatment, neurological function impairment score was performed; the infarct area ratio was determined by 2,3,5-tripheyl tetrazolium chloride (TTC) staining; apoptosis of brain cells was observed by TUNEL method; the expressions of Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry.Results: Compared with the blank control group and the sham operation group, the neurological function impairment score, cerebral infarct area ratio, apoptosis, and the expressions of Bax and Caspase-3 in the model group were significantly increased, while the expression of Bcl-2 was significantly decreased, and there were significant between-group differences (allP<0.05). After the treatment, there were statistically significant differences among the treatment groups in the neurological function impairment score, cerebral infarct area ratio and apoptosis in the ischemic side of rats, as well as the expressions of Bcl-2, Bax and Caspase-3 (allP<0.05), and from the figures, tables and statistical analysis, it was found that a better tendency in the acupuncture plus mild hypothermia group than the acupuncture group or mild hypothermia group.Conclusion: Acupuncture plus mild hypothermia can protect the brain cells by improving neurological function impairment, decreasing cerebral infarct area ratio, reducing the number of apoptotic cells in the ischemic area and regulating the expressions of apoptosis related proteins to inhibit apoptosis.

Objective:To observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury (CIRI), and the influence on protein expression levels of phosphorylated Raf-1, MEK-2 and ERK1/2 in the mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinases (ERK) pathway, and to explore the mechanism of acupuncture plus mild hypothermia therapy for the ischemic stroke. Methods:Ninety Sprague-Dawley (SD) rats were randomly divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, 15 rats in each group. Except the rats in the blank control group, the remaining rats were used to prepare the middle cerebral artery occlusion (MCAO) models according to the modified occlusion method using lines, while only the occlusion lines were inserted without blocking the brain arteries of rats in the sham operation group. When the vital signs of rats were stable, rats in the blank control group did not receive any intervention; rats in the sham operation group and the model group received fastening without treatment; rats in the acupuncture group, the mild hypothermia group, and the acupuncture plus mild hypothermia group were treated with the corresponding therapeutic methods. 72 h later, observed neurologic injury score, evaluated infarction area ratio by 2,3,5-tripheyl tetrazolium chloride (TTC) staining, determined apoptosis by TUNEL assay, and measured the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels in rat ischemic hippocampal tissues by Western blot assay. Results: Compared with the blank control group and the sham operation group, after modeling, the neurologic injury score, infarction area ratio and apoptotic cells were increased, and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly increased in the model group; the differences were statistically significant (P<0.05 orP<0.01). Compared with the model group, after acupuncture or mild hypothermia therapy, neurologic injury score and infarction area ratio were decreased; apoptotic cells and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly decreased; the differences were statistically significant (P<0.05 orP<0.01). Compared with the acupuncture group, neurologic injury score and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were decreased in the acupuncture plus mild hypothermia group; differences between the groups were statistically significant (P<0.05 or P<0.01). Compared with the mild hypothermia group, phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels decreased in the acupuncture plus mild hypothermia group, and differences were statistically significant (P<0.01). Conclusion:Acupuncture or mild hypothermia therapy can improve neurologic injury, reduce infarction area and apoptosis, which brought about protective effect on the brain tissues, in the MCAO model. The protective effect of acupuncture plus mild hypothermia group is the strongest. The mechanism may involve the MAPK/ERK pathway, by reducing the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels.

BACKGROUND:Preliminary studies of our research group mainly focus on the role of nucleus of solitary tract in gastric noxious stimulation and acupuncture point, while dorsal nucleus of vagus nerve, nucleus of solitary tract and area postrema are the necessary component for vagus nerve complex, and dorsal nucleus of vagus nerve plays a crucial role in the regulation of gastric functions. OBJECTIVE:To observe the effect of acupuncture at Zusanli, Neiguan, Pishu and Shenshu acupoints on the gastric-related neurons in dorsal motor nucleus of the vagus nerve. METHODS:The gastric related neurons in dorsal motor nucleus of the vagus nerve of gastric distension rats were identified using extracellular recording technique, according to neuroelectrophysiology method. Acupuncture stimulation was given to Zusanli, Neiguan, Pishu and Shenshu for 15 seconds. The effect of body stimulation on the gastric distension related neurons in dorsal motor nucleus of the vagus nerve of rats was recorded. RESULTS AND CONCLUSION:Among the involved 24 rats, 90 neurons at paraventricular hypothalamic necleus exhibited discharge. Among the discharged neurons, 54 neurons were related to gastric distension. After acupuncture stimulation at Zusanli, Neiguan, Pishu and Shenshu, there were 44, 39, 38, 27 neurons giving a response, the reaction rate was 82%, 72%, 70%and 50%, respectively. Acupunture at different nerve segment acupoints can activate the dorsal motor nucleus of the vagus nerve to different degrees, which is involved in the regulation of gastric motility.

BACKGROUND:Previous studies concerning neuromechanism of acupuncture for regulating gastric function mainly focused on peripheral nerve, involving in central nervous system. In particular, there are few studies addressing higher central nervous system.
OBJECTIVE:To explore the effects of acupuncture at Neiguan (PC6) and Zusanli (ST36) on gastric-related neurons in the hypothalamic paraventricular nucleus.
METHODS:Using microelectrode extracellular recording technique, gastric related neurons in the paraventricular nucleus were found in gastric distension rats. Stimulation was conducted for 30 seconds by hand acupuncture at Neiguan and Zusanli. The effects of acupuncture on gastric-related neurons in the paraventricular nucleus were observed.
RESULTS AND CONCLUSION:The discharge of 109 neurons in the hypothalamic paraventricular nucleus of 60 rats was recorded. A total of 56 gastric-related neurons of the 109 neurons were observed. The number of neurons response to acupunctures at Zusanli, Neiguan, Pishu (BL20) and Weishu (BL21) were respectively 44, 47, 29, and 33, with the reaction proportion of 78.57%, 83.47%, 51.79%and 58.93%. Results suggested that there exist gastric distension stimulation and acupuncture stimulations from neurons soma to visceral reaction in the hypothalamic paraventricular nucleus, which simultaneously participates in the regulation of acupuncture on the gastric function.

PurposeTo investigate the formation of neovascularization using dynamic contrast-enhanced MRI (DCE-MRI) in C6 glioma margin in rats, and to explore its utility.Materials and MethodsC6 glioma model was created in 24 SD rats. Routine and DCE-MRI was performed 4 days, 8 days, 12 days, 16 days, 20 days and 24 days after transplantation. Pathology and immunohistochemistry exams were also obtained to evaluate marginal tumor cells and neovascularization.ResultsNeovascularization was observed in C6 glioma margin within (257.658±53.663)μm range. Vessel co-option index difference was statistically significant (P<0.01). Tumor cells surrounding the co-opted vessels were strongly positive on proliferating cell nuclear antigen and matrix metallopeptidase 2 staining. Vessel co-option regression and transformation were observed on day 16 on Tenascin-C andα smooth muscle actin staining. For DCE-MRI analysis, Ktrans and Kep were higher on day 12 and 16 (P<0.05). There was significant negative correlation between co-option index and Ktrans (r=-0.9243 and-0.8347,P<0.05). Vp showed no statistically significant difference at all time points (P>0.05).Conclusion The neovascularization in tumor margin demonstrates vessel co-option. DCE-MRI parameters Ktrans and Kep are negatively correlated with co-option index, indicating that Ktrans and Kep could be a well-defined imaging-based biomarker for glioma neovascularization.

Proteomics, which has been widely used in life science, is an emerging discipline following genomics. It can help to explore the pathogenic mechanism and early onset marker of Bacillus anthracis, playing an important part in the prevention, diagnosis and treatment of B.anthracis. In this paper,the application of proteomics in the research of B.anthracis is reviewed.

Objective: To analyze the association between alcohol consumption and hyperuricemia among residents in Chengdu City, so as to provide the evidence for prevention and control of hyperuricemia. Methods: Based on the Natural Cohort Study in Southwest Area, residents at ages of 30 to 79 years were recruited in Chengdu City in 2018. Information of demographics, smoking, alcohol consumption and diet were collected through a questionnaire survey. Blood uric acid was tested in the laboratory. Participants were divided into never, moderate and excessive drinking groups based on alcohol consumption. A multivariable logistic regression model was used to analyze the association between alcohol consumption and hyperuricemia, and subgroup analysis was conducted according to gender, current residence, physical activity and body mass index (BMI). Results: A total of 20 164 residents were investigated, including 8 776 males (43.52%) and 11 388 females (56.48%), with a mean age of (51.22±12.33) years. There were 9 769 never-drinkers (48.45%), 8 310 moderate-drinkers (41.21%), and 2 085 excessive-drinkers (10.34%). Hyperuricemia was detected in 4 101 patients, with a detection rate of 20.34%. Multivariable logistic regression analysis showed that moderate drinking (OR=1.122, 95%CI: 1.031-1.222) and excessive drinking (OR=1.529, 95%CI: 1.349-1.734) were associated with an increased risk of hyperuricemia. Moderate and excessive drinking were associated with an increased risk of hyperuricemia among men, urban residents, residents with a high level of physical activity, and those with BMI less than 24 kg/m2 (all P<0.05). Excessive drinking were associated with an increased risk of hyperuricemia among rural residents, residents with a low level of physical activity and with BMI of 24 kg/m2 and higher (all P<0.05). Conclusions Both moderate and excessive drinking are associated with an increased risk of hyperuricemia. Moderate drinking is not associated with a higher risk of hyperuricemia among rural residents, residents with a low level of physical activity and with BMI of 24 kg/m2 and higher.