BACKGROUND: Electroacupuncture has a good clinical effect on Alzheimer disease, but its mechanism remains unclear.OBJECTIVE: To observe the effect of electroacupuncture on number of synaptic numeric density (Nv), surface density (Sv) and average size of synaptic conjunction and ultrastructure in hippocampal CA3 area of neurons of model rats with Alzheimer disease (AD).DESIGN: Completely randomized grouping and controlled study.SETTING: Department of Traditional Chinese Medicine of Sichuan Provincial People's Hospital; College of Acupuncture & Moxibustion and Massage of Chengdu University of Traditional Chinese Medicine.MATERIALS: A total of 50 male SD rats of 24-month old weighing (480±20) g and 6 male rats of 3-month old weighing (250±15) g were selected in this study. Passage water maze (2.1×1.7×0.6) m3 was made of black glasses with 40 cm deep water and 4 caecums. WQ1002F Hans electrically heated distilling apparatus was used.METHODS: The experiment was carried out at the Grade Ⅲ Animal Experimental Center of Chengdu University of Traditional Chinese Medicine between, September 2002 and June 2003. ① Old SD rats were grouped on the basis of water-maze test results. Firstly, 6 young rats were submitted to water-maze test at 8 days before modeling for 4 continuous days to obtain average escaping latency. Secondly, 50 old rats were accepted in the water-maze test at 4 continuous days before modeling to obtain average escaping latency. A total of 36 rats whose latencies were less than average values plus one standard deviation of young rats were regarded as normal old rats. Among them, 12 rats were randomly divided into control group and sham operation group with 6 in each. Another 24 rats were transected at fornix-fimbria AD modeling. Thirdly, 24 modeling rats were all adopted in the water-maze test at 2 days after modeling. Rats whose latencies were more than average value plus two standard deviation of young rats were chosen as AD model. Twelve AD models were randomly divided into model group and electroacupuncture group with 6 in each.② On the 6th day after modeling, rats in electroacupuncture group were acupunctured at Baihui (Du20) of 0.5 inch in slope, Yongquan (K1),Taixi (K3) and Xuehai(Sp10) of 0.3 inch in depth with No. 30 milli-needle (3.33 cm); Then, electrically heated distilling apparatus was used with successive waves, 20 Hz in frequency and 2-4 V in voltage. Tolerant stress of rats at quiet state was regarded as the standard value (2 mA),the needle was maintained for 30 minutes, and the acupuncture was done once a day for totally 20 successive days. Rats in control group sham operation group (cerebral cortex was exposured at the same site as model rats, and fornix-fimbria was not cut off) and model group were only fixed but treated with nothing. ③ After treatment, ultrastructure in hippocampal CA3 area of rats was observed with transmission electron microscope;synapse numbers and cross-point numbers between synaptic conjunction and test line were counted with stereological technique; stereological parameters, such as numeric density (Nv), area surface (Sv) and average size of synaptic conjunction, which could reflect plastic changes of synaptic form, were calculated at the same time. ④ Differences between every two groups were compared with t test at regular variance and t'test at irregular variance.MAIN OUTCOME MEASURES: Comparisons of numeric density (Nv),surface density (Sv) and average size of synaptic conjunction.RESULTS: A total of 24 old rats were involved in the final analysis with 6 in each group. ① Ultrastructure: Synaptic density of control and sham operation group was higher than that of model group, and average area of synapse was smaller;, synaptic density of electroacupuncture group increased compared with that of model group, and average area of synapse was smaller. ② Numeric density and surface density in hippocampal CA3 area in model and electroacupuncture group were lower than those of control group and sham operation group (P ＜ 0.05). There was no significant difference between sham operation group and control group (P ＞ 0.05), and those of electroacupuncture group were higher than those of model group (P ＜ 0.01). ③ Average size of synaptic conjunction in hippocampal CA3 of model and electroacupuncture group was higher than that of control and sham operation group (P ＜ 0.05). There was no significant difference between sham operation group and control group (P ＞ 0.05), and the value of electroacupuncture group was lower than that of model group (P ＜ 0.01). CONCLUSION: Electrotherapy can repair synaptic form and inhibit synaptic degeneration of hippocampal neurons in AD rats.

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

OBJECTIVE: To explore the neuroprotective effect of electroacupuncture therapy on damaged cholinergic neurons in hippocampus in aged rats with Alzheimer's disease (AD). METHODS: Thirty six aged male rats were randomly divided into normal control group, sham-operated group, untreated group and electroacupuncture group. Animal model of AD was established with fimbria-fornix transection. The rats in the electroacupuncture group received electroacupuncture on Baihui (DU 20), Yongquan (KI 1), Taixi (KI 3) and Xuehai (SP10). The activity of choline acetyltransferase (ChAT) in septal area of brain was detected by radioimmunoassay, and the protein expressions of nerve growth factor (NGF) and c-fos in CA3 region of hippocampus were detected by immunohistochemical assay. RESULTS: The ChAT activity and the expression levels of NGF and c-fos proteins in the electroacupuncture group were significantly higher than those in the untreated group. CONCLUSION: Electroacupuncture therapy can protect cholinergic neurons in hippocampus in aged rats with AD by means of promoting synthesis of c-fos protein and increasing the expression level of NGF.

Objective:To investigate the current status of cognitive frailty among elderly patients in Urumqi and to identify its influencing factors.Methods:From March to December 2019, the elderly from 3 tertiary hospitals′ geriatrics centers in Urumqi were recruited using the general information questionnaire, FRAIL Scale, Montreal Cognitive Assessment and Clinical Dementia Rating.Results:A total of 1 006 elderly patients were surveyed, among which, 131(13.0%) cases were deemed to have developed cognitive frailty. The results of multivariate logistic regression analysis showed that age, depression, Instrumental Activities of Daily Living Scale score and diabetes were influencing factors of cognitive frailty ( P<0.001). Conclusion:The prevalence rate of cognitive frailty in elderly patients is relatively high. Medical staff should attach great importance to the assessment of cognitive frailty in elderly patients and take targeted intervention in time to prevent, slow down or reverse the onset and development of cognitive frailty.

Objective: To observe the therapeutic effect of sinew-regulating bone-setting manipulations for knee osteoarthritis (KOA) model rabbits and its impacts on the chondrocyte apoptosis rate and the levels of interleukin (IL)-1β and nitric oxide (NO). Methods: According to the random number table method, 30 New Zealand white rabbits were divided into a normal group (n=9) and a modeling group (n=21). Rabbits in the modeling group were used to establish KOA models with the modified Hulth method. At the 8th week, three rabbits were sacrificed to verify the model and the remaining 18 rabbits were randomly divided into a model group (n=9) and an intervention group (n=9). Rabbits in the normal group and model group were bred routinely without any intervention. Rabbits in the intervention group were treated with the sinew-regulating bone-setting manipulations, 10 min/time, once every other day for a total of 20 times. The Lequesne MG knee function rating was used to evaluate the behavioral differences of the rabbits in each group. The Pelletier score was used to evaluate the general changes of the rabbits. The Mankin score was used to evaluate the pathology of knee cartilages. The enzyme-linked immunosorbent assay and nitrate reductase methods were used to determine the levels of IL-1β and NO in serum and synovial fluid of each group, respectively. In situ terminal deoxynucleotidyl transferase-mediated nick and labeling method was used to determine the apoptosis of chondrocytes in each group. Results: Compared with the normal group, the scores of Lequesne MG, Pelletier and Mankin, and the levels of IL-1β and NO in the model group were increased (P<0.05), which indirectly indicated the success of the model. Compared with the model group, the scores of Lequesne MG, Pelletier and Mankin, IL-1β and NO levels, and chondrocyte apoptosis rate of the intervention group were decreased, and the differences were statistically significant (P<0.05). Conclusion: The sinew-regulating bone-setting manipulations can reduce the levels of IL-1β, NO, and chondrocyte apoptosis rate, and delay the articular cartilage degeneration, therefore, having a good therapeutic effect on KOA.

OBJECTIVETo observe the effects of electroacupuncture (EA) on circadian rhythm of temperature and melatonin (MT) in depression rats model induced by chronic stress, so as to explore the biological mechanism of EA for depression.

METHODSTwenty-four SD rats were randomly divided into a control group, a model group and an EA group, 8 cases in each one. Rats in the control group were treated with normal diet for 21 days without any treatment. In the model and EA group, rat model was established by chronic unpredictable stress combined with solitarily feeding method, and rats in the EA group was treated with EA at "Baihui" (GV 20), "Yintang" (GV 29) 1 h before stress stimulation everyday, 2 Hz in frequency and intensity was favorable with the head of rat slightly shivering. The needles were retained for 20 min, once a day for totally 21 days. After EA treatment, open-field experiment was adopted to observe the behavioral improvement; the rats temperatures were monitored at six time points (2:00, 6:00, 10:00, 14:00, 18:00, 22:00) and orbital blood sampling was collected. The level of serum MT was tested by enzyme linked immunosorbent assay. The circadian rhythm changes of temperature and serum MT in each group were compared.

RESULTSThe numbers of horizontal movement and vertical movement in the model group were obviously lower than those in the control group (both P < 0.05), while those in the EA group were significantly improved compared with those in the model group (both P < 0.01). The circadian rhythm of temperature and MT disappeared in the model group, which was improved into normal level after EA treatment.

CONCLUSIONThe electroacupuncture has regulation effects on circadian rhythm of temperature and melatonin in depression rat model induced by chronic stress.

Animals ; Circadian Rhythm ; Depression ; metabolism ; physiopathology ; therapy ; Electroacupuncture ; Humans ; Male ; Melatonin ; metabolism ; Rats ; Rats, Sprague-Dawley

This study was aimed to apply the electronic nose (E-nose) in the research of traditional Chinese medicine (TCM). The discussion was made on difficulties of using E-nose. The solution plan was proposed and the discrimination model was established. It provided a simple, rapid and effective analysi method in the identification of TCM. It also provided new ideas for the research and application of gas sensor arrays. E-nose was used in the ex-traction of TCM scent characteristics. Based on ion mobility spectrometry of MOS sensor, the fingerprint of TCM scent was established. The maximum response value of the sensor was used as analysis index. According to the diffi-culties of identification, two solution plans were proposed. Firstly, different detectors were employed to complete the classification. Secondly, radial basis function (RBF) and random forests (RF) were combined and then a cascade classifier was constructed in order to achieve the maximum of information obtained in conditions where the number of measurements, metal oxide semiconductor sensors in E-nose was limited. The results showed that both plans were accurate and practical with relatively high upper correct judge rate and better cross-validation (The highest upper correct judge rates were 95% and 100%, 96% and 80%, respectively). It was concluded that this study firstly ap-plied cascade classifier in the establishment of TCM identification by E-nose. With limited amount of sensors, the maximum information was received through data mining. Using E-nose in the identification of TCM was rapid and accurate. The established pattern recognition method was maneuverable with accurate identification rate and stability compared to conventional sensory identification method. It provided a simple and rapid analysis method for the iden-tification of TCM.

This study assessed the clinical application of transvaginal three-dimensional ultrasound (3D TVUS) in the diagnosis of congenital uterine malformation. A retrospective study was performed on 62 patients with congenital uterine malformation confirmed hysteroscopically and/or laparoscopically. The patients were subjected to transvaginal two-dimensional ultrasound (2D TVUS) and 3D TVUS. The accuracy rate was compared between the two methods. The accuracy rate of 3D TVUS was (98.38%, 61/62), higher than that of 2D TVUS (80.65%, 50/62). 3D TVUS coronal plane imaging could demonstrate the internal shape of the endometrial cavity and the external contour of the uterine fundus. It allowed accurate measurement on the coronary plane, and could three-dimensionally show the image of cervical tube, thereby providing information for the diagnosis of some complex uterine malformation. 3D TVUS imaging can obtain comprehensive information of the uterus malformation, and it is superior to 2D TVUS for the diagnosis of congenital uterine malformations, especially complex uterine anomaly.

Objective:To observe the effect of liver-soothing and mind-regulating acupuncture method on the resting-state electroencephalography (EEG) in rats with post-traumatic stress disorder (PTSD),and to provide evidence for the effect mechanism study and clinical application of acupuncture intervention for PTSD.Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group,a model group,a grasping group,a paroxetine group and an acupuncture group,with 12 rats in each group.Except for rats in the blank control group,rats in the other groups were subjected to preparing the PTSD models using 'incarceration plus electric shock' method.After interventions,changes in rat behavior of each group were observed;changes in resting-state EEG were collected and analyzed with multichannel EEG acquisition and analysis system,and image analysis and statistical processing were performed.Results:Compared with the blank control group,the average escape latency in the model group was significantly longer (P＜0.05),and the times of crossing the platform and the effective areas were all significantly reduced (P＜0.01).Compared with the grasping group,the average escape latencies in the paroxetine group and acupuncture group were significantly shortened (P＜0.05),and the times of crossing the platform and the effective areas were all significantly increased (P＜0.05).There were no significant differences in the average escape latency,the times of crossing the platform and the effective areas between the acupuncture group and paroxetine group (all P＞0.05).Compared with the blank control group,the α-wave power spectrum value in the model group was significantly decreased,and the power spectrum values of β-wave,δ-wave and a-wave were significantly increased (all P＜0.01);compared with the grasping group,α-wave power spectrum values in the paroxetine group and acupuncture group were significantly increased (both P＜0.01),and the power spectrum values of β-wave,δ-wave and a-wave were decreased significantly (all P＜0.01).The power spectrum values of α-wave,β-wave,δ-wave and (e)-wave of rats in the acupuncture group were not significantly different from those in the paroxetine group (all P＞0.05).Conclusion:Liver-soothing and mind-regulating acupuncture method can significantly improve the abnormal EEG activity in PTSD rats,which may be one mechanism of liver-soothing and mind-regulating acupuncture method in effectively affecting the brain function in PTSD rats.

Objective: To study the effect of oxymatrine on oxidative stress and inflammatory reaction after cerebral ischemia reperfusion in rat. Methods: A total of 140 SD rats were randomly divided into the sham group, model group and oxymatrine low, medium, high-dose groups (28 in each group). Beyond the sham group, the other groups were established the rat model of focal cerebral ischemic reperfusion with Zea-Longa occluded suture. The the low-, medium- and high-dose matrine groups were given 25, 50, and 100 mg/kg oxymatrine solutions injected intraperitoneally from the second day after operation for 7d. , The model group and the sham group were injected with an equal volume of saline. The neurological deficits, ratio of infarct volume, water content of the brain were evaluated; the morphological changes of brain tissue were observed by HE Staining, and the cell apoptosis were observed after TUNEL staining; the activity of antioxidant enzymes and malondialdehyde (MDA) in brain tissue were measured by biochemical analysis; the content of NO and the activity of iNOS were measured; the expression of NF-ĸB were determined by Western blotting. The inflammatory cytokines in brain tissue were determinated. Results: Compared with the model group, the neurological function score, brain tissue water content, infarct volume and apoptotic index decreased in the rats of oxymatrine medium-, high-dose groups (P<0.05 or P<0.01). The activity of SOD (148.68 ± 9.12 U/mg, 161.34 ± 10.09 U/mg vs. 140.63 ± 8.47 U/mg), CAT (3.90 ± 1.32 U/mg, 4.15 ± 1.47 U/mg vs. 2.73 ± 0.89 U/mg), GSH-Px (11.46 ± 2.65 U/mg, 13.59 ± 3.27 U/mg vs. 9.35 ± 2.03 U/mg) were significantly increased (P<0.05 or P<0.01). The content of MDA (20.18 ± 3.59 μmol/g, 17.46 ± 3.21 μmol/g vs. 29.86 ± 5.40 μmol/g), iNOS (12.64 ± 1.83 U/mg, 11.75 ± 1.62 U/mg vs. 14.17 ± 2.06 U/mg), NO (23.64 ± 2.18 μmol/g, 21.27 ± 2.03 μmol/g vs. 27.82 ± 2.42 μmol/g), TNF-α (43.9 ± 6.3 nmol/L, 37.2 ± 5.8 nmol/L vs. 56.3 ± 6.9 nmol/L), IL-1β (715.0 ± 68.5 nmol/L, 683.9 ± 70.8 nmol/L vs. 930.8 ± 91.4 nmol/L), IL-6 (168.4 ± 22.5 nmol/L, 133.7 ± 18.1 nmol/L vs. 212.5 ± 24.7 nmol/L) were significantly decreased (P<0.05 or P<0.01). The expression of NF-ĸB in brain (0.35 ± 0.13, 0.24 ± 0.08 vs. 0.62 ± 0.15) were significantly down-regulated (P<0.05 or P<0.01). Conclusions The Oxymatrine inhibits oxidative stress injury and inflammatory response after cerebral ischemia-reperfusion in rats.