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.

Objective To investigate the expression and tissue distribution of CD31 in the process of the calf cortical with partial cancellous bone xenograft, to explore the healing mechanism and relationship between bone remodeling and revascularization, and to provide a basis for the further study of tissue engi-neered bone.Methods Thirty-six New Zealand rabbits were selected as our experimental animals.All ani-mals were implanted with the calf cortical with partial cancellous bone.Specimens were procured at 4, 8, 12 and 24 weeks after surgery, respectively.The expression and tissue distribution of CD31 were observed in the process healing with general condition, X-ray, histology and immunohistochemistry.The image analy-sis computer system was used for quantitative analysis.The extent of revascularization and remodeling was separately represented by revascularization indexes and new bone areas.The relationship was studied be-tween bone remodeling and revascularization.Results ⑴ The X-ray showed significant healing.⑵ The positive expression of CD31 was seen in the whole process of bone xenograft and was different at various stage and sites.By the 4 weeks and 8 weeks postoperative,the strong positive expression was seen in the pe-ripheral soft tissue of graft bone, the callus of joint and the periphery of cartilage nests.Much positive ex-pression was seen in the cancellous bone.There was seen some single, cord, and cluster vascular endotheli-al cells.The positive expression was seen in the enlarged haversian canals and periphery.By the 24 weeks postoperative,the positive expression was seen in the enlarged haversian canals and periphery with increasing newborn capillaries.⑶ The correlation analysis between new bone areas and revascularization showed a closely positive correlation ( r =0.984).Conclusions The positive expression of CD31 was seen in the whole process of bone xenograft and was different at various stage and sites.The positive expression of CD31 reflected the extent of revascularization and bone remodeling of bone grafts.The extent of revascularization of bone grafts and bone remodeling showed a closely positive correlation.

Objective To evaluate the incidence of surgical site infection (SSI)in patients with colorectal cancer and identify the risk factors of SSI.Methods A total of 392 patients who underwent colorectal cancer resection at the First Affiliated Hospital of Chongqing Medical University between September 2012 and September 2013 were included and analyzed in terms of the presence of SSI.SSI risk factors were identified by both univariate and multivariate analysis.Results The overall incidence of SSI was 23.0%.Univariate analysis showed that duration of operation above 75 th percentile,colostomy,surgical procedure, type of surgical incision,and American Society of Anesthesiologists (ASA)score were significantly associated with higher risk of SSI (P <0.05).Binary logistic regression suggested that duration of operation above 75 th percentile (P = 0.000,OR =3.017),colostomy (P =0.008,OR=2.642),Contaminated incision (P =0.016,OR=3.311)and laparoscopic surgery (P =0.016,OR=0.523)were significantly different in terms of presence or absence of SSI.Conclusions Duration of operation above 75th percentile,colostomy and contaminated incision are independent risk factors,while laparoscopic surgery is a protective factor for SSI in patients with colorectal cancer.

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 assess the potential of whole blood IFN-γassay for diagnosing mycobacterium in rhesus macaques.Methods Firstly, basic serum IFN-γconcentrations of TST-negative and -positive rhesus macaques were detected.Then, heparinized whole blood from TST-negative and-positive rhesus macaques was incubated with PBS and 200 IU bovine-PPD ( tuberculin purified protein derivative ) for about 24 h, respectively.The supernatant plasma were harvested and used to determine the concentrations of IFN-γ.The results of plasma IFN-γconcentrations and stimulation index ( SI) were used to analyze the diagnostic potential of the whole blood IFN-γassay.Results The basic serum concentrations of IFN-γfor the TST-positive monkeys were significantly higher than that of the TST-negative macaques, showing a high coefficient of variation.There was no significant effect on the production of IFN-γin the TST-negative macaques.While significantly elevation of IFN-γconcentrations was found in stimulated plasma of TST-positive macaques (P<0.01).The SI of TST-positive macaques was significantly higher than the TST-negative ones.ROC curve analysis revealed that IFN-γconcentrations and SI could be used as evaluation index of whole blood IFN-γassay.Conclusions Based on a small sample experiment we have demonstrated that whole blood IFN-γassay may be one possible auxiliary diagnostic method for tuberculin skin test.

OBJECTIVETo establish the steriod-induced osteoporosis model with the type of Kidney-Yin deficiency.

METHODSTotally 45 female Kunming mice were randomly divided into normal group,model group and Liuwei Dihuang pills(Chinese character: see text)group. The model was established by intramuscular injecting of Dexamethasone. Liuwei Dihuang pills (Chinese character: see text) group was administered orally with Liuwei Dihuang pills (Chinese character: see text). The signs and symptoms of mice were observed dynamically. All the animals were sacrificed at the end of the 6th weeks. The level of ACTH, cAMP, cGMP, TSH and E2 in serum were detected to evaluate deficiency of Kidney-Yin. Morphological changes and bone density were observed to evaluate osteoporosis.

RESULTS(1) Compared with control group, mice in model group appeared obvious Kidney-Yin deficiency symptoms, including hair dry, restlessness, excitability, hard stool, and yellow. (2) Compared with control group,the weight of mice in model group gained slower (P<0.01); the index of adrenal gland,liver and spleen decreased (P<0.01, P<0.01 ,P<0.01); the level of ACTH and TSH increased (P<0.01 ,P<0.01), the level of E2 decreased (P<0.01) and the ratio of cAMP/cGMP increased (P< 0.05). (3)Compared with control group,the bone density of lumbar vertebra and femur in model group were significantly decreased (P<0.01, P<0.05); HE staining revealed osteoporosis in model group mice. (4)However, the Liuwei Dihuang pills (Chinese character: see text) group can partly antagonize the inhibition of the HPA axis, alter the disordered sex hormone and the ratio of cAMP/cGMP, and reverse the osteoporosis partly.

CONCLUSIONthe model of osteoporosis with type of Kidney-Yin deficiency could be established by Dexamethasone intramuscular injection. With less interference, it wight be a stable and reliable modeling method for integration of disease and syndrome in TCM.

Animals ; Bone Density ; Dexamethasone ; toxicity ; Disease Models, Animal ; Female ; Kidney Diseases ; etiology ; Medicine, Chinese Traditional ; Mice ; Osteoporosis ; chemically induced ; Yin Deficiency ; complications

Objective To use the matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) technique for detecting the mutation gene of neonatal non-syndromic hereditary hearing impairment gene in Guangxi and to investigate its effectiveness and feasibility in clinical application.Methods A total of 7 100 newborns were performed the hearing preliminary screening and secondary screening by adopting AABR.The genomic DNA was extracted by the heel blood spot.Twenty mutation characteristics of 4 deaf predisposing genes were detected by MALDI-TOF-MS.Results The pass rate of hearing screening in 7 100 newborns was 97.11％ (6 895/7 100),the positive rate of neonatal gene mutation was 3.54％ (251/7 100),in which the GJB2 gene mutation was in 131 cases,the carrying rate was 1.84％,235delC heterozygous mutation was in 108 cases.SLC26A4 gene mutation was in 93 cases,which dominated by 1229C＞T heterozygous mutation and IVS7-2A＞G heterozygous mutation,mtDNA12SRNA gene mutation was in 16 cases and GJB3 gene mutation was in 11 cases.Conclusion Adopting the MALDI-TOF-MS screening technique can increase the detection rate of hot point mutation in common deaf related genes and discover neonatal genetic NSHI from molecular level and provides the corresponding geneticconsulting guidance for early finding and predicting deaf occurrence,and formulating the interventional measures.

?AlM: To investigate the expression of miR-181 in the lens tissue of cataract and the regulating mechanism of miR-181 on apoptosis of human lens epithelial cell.
?METHODS:Real time q-PCR was used to measure the expression of miR-181 in the anterior lens capsules of age - related cataract and human lens epithelial cell apoptosis model. miR- 181 mimic and inhibitor were transfected using Lipofectamine 2 000 to regulate the expression of miR-181, and then Real time q-PCR was used to verify transfection efficiency. Flow cytometry was used to detect the change of cell apoptosis rate.
? RESULTS: Compared with control group, the expression of miR-181 was significantly higher in both the anterior lens capsules of age-related cataract and human lens epithelial cell apoptosis model; the relative expression of miR-181 in lens epithelial cells transfected with miR-181 mimic was increased, whereas decreased in cells transfected with miR-181 inhibitor;the apoptosis rate of cells transfected with miR - 181 mimic was increased, while reduced in miR-181 inhibitor group. Each result was statistically significant (P<0. 01).
?CONCLUSlON:High expression of miR-181 is detected in anterior lens capsule of age-related cataract. miR-181 might play a certain role in the pathogenesis of cataract via promoting human lens epithelial cell apoptosis. miR-181 probably becomes a new approach for the nonoperative treatment of cataract, but the concrete mechanism still needs to be further studied.

Objective To investigate the effects of different combinations of vitamin (Vit) C, Vit E and GSH on the biochemical markers and bone mineral density (BMD) in ovariectomized rats. Methods Seventy female SD rats aged 4 months were divided into two groups, 20 rats with sham operation in control group and 50 rats with bilateral oophorectomy in model group. 3 months later, 10 rats in the model group and 10 rats in the control group were randomly selected and their body weight, uterus weight, BMD of the left femurs and lumbar spines,biomeehanical characteristics of the left femurs, serum levels of Ca2+ , creatinine (Cr), alkaline phosphatase (ALP),superoxide dismutase (SOD) , malondialdehyde (MDA) , GSH-peroxidase (GSH-Px) and serum resisting abilities to OH- were determined. Then the rest rats were divided into five groups: A (sham), B (OVX control), C (Vit C +Vit E), D (GSH) and E (Vit C +Vit E +GSH). Vit C, Vit E and GSH were given 750rng/kg, 250 mg/kg, and 125 mg/kg, respectively daily for 3 months. And then the biochemical markers and BMD were measured. Results 3 months after treatment with antioxidants, BMD of left femurs and lumbars spines was increased, while the level of serum ALP was decreased markedly in B, C and D group as compared with that in B group. The serum level of SOD, GSH-Px and serum resisting ability to OH- were increased in D and E groups and the level of MDA decreased in C and D groups as compared with that in B group. Conclusion Vit C, Vit E and GSH increased BMD, prevented the decrease of SOD and GSH-Px and elevated serum resisting ability to OH-in ovariectomized rats.

Objective To investigate the effects of different combinations of antioxidants VitC, VitE and GSH on the biomechanical and biochemical markers in ovariectomized rats, Methods Seventy female SD rats, aged 4 months, were divided into 2 groups (control and model) randomly. Fifty rats in the model group underwent ovariectomy, while the other 20 rats in the control group had sham operations. Three month slater, 10 rats in the model group and 10 rats in the control group were selected randomly to detect their body weight, uterus weight, BMD of the left femur and lumbar vertebra, biomechanical characteristics of the left femur, levels of serum Ca2 +, Cr, ALP, SOD, MDA and GSH-Px and serumal resistance to OH<'->. Then the rest were divided into 5 groups: A (sham), B (model), C (VitC + VitE), D (GSH), and E (VitC + VitE +GSH). VitC, VitE and GSH were given in 750 mg/kg, 250 mg/kg, and 125 mg/kg every day for 3 months,respectively. And then the biomechanical and biochemical markers were measured. Results Three months after ovariectomy, the body weight of the rats in the model group increased markedly compared with the control group, while BMD of the left femur and lumbar vertebra, biomechanical maximal load and uterus weight decreased. The serum levels of Ca<'2+>, ALP and Cr increased. Three months after antioxidant treatment,the biomechanical maximal load and elastic load of the left femur and the maximal load of the 5th lumbar vertebra, the serum levels of SOD, GSH-Px and the serumal resistance to OH<'-> in groups D and E increased markedly, while the serum level of MDA decreased in groups C and D and the level of serum ALP decreased in all the treatment groups. Conclusion GSH and combination of VitC, VitE and GSH play a positive role in treatment of osteoperesis in ovariectomized rats.