Objective To evaluate the relationship between extracellular signal-regulated kinase (ERK)and ketamine-induced apoptosis in rat hippocampal neurons.Methods Sprague-Dawley rats at 18 days of gestation were anesthetized.The fetal rats were obtained under the sterile condition and decapitated.The hippocampal neurons were isolated and primarily cultured for 5 days,and were seeded in 6-well plates (2 ml/well) or in 96-well plates (100μl/well) at a density of 5 × 105/ml.The cells were randomly divided into 4 groups (n =18 each):control group (group C),fibroblast growth factor (FGF-2,an ERK agonist) group (group F),ketamine group (group K) and FGF-2 + ketamine group (group FK).The cells were cultured in the plain culture medium in group C.FGF-2 50 ng/ml was added to the culture medium in group F.Ketamine was added to the culture medium in group K.FGF-2 50 ng/ml was added to the culture medium at 20 min before ketamine 100 μmol/L was added in group FK.The phosphorylation of ERK in hippocampal neurons was detected by Western blot at 10 min after treatment.At 24 h after treatment,the neuronal apoptosis was detected by Hoechst33342/PI staining,and the cell survival rate was detected by MTT assay.The apoptosis rate was calculated.Results Compared with group C,the phosphorylation of ERK in hippocampal neurons and the cell survival rate was significantly decreased and the apoptosis rate was increased in K and FK groups (P ＜ 0.05).There was no significant difference in the parameters mentioned above between F and C groups (P ＞ 0.05).The phosphorylation of ERK in hippocampal neurons and the cell survival rat was significantly higher and the apoptosis rate was lower in group FK than in group K (P ＜0.05).Conclusion Ketamine induces apoptosis in rat hippocampal neurons by inhibiting activation of ERK in hippocampal neurons.

Objective To investigate the changes of expression of stromal cell-derived factor 1α (SDF-1α) in retinas after partial optic nerve injury in rats. Methods Models with injury of partial optic nerve were induced in rats. Retinal tissues were collected 1,2,3,5,7,10 and 14 d after injury. Enzyme linked immunosorbent assay and Real-time quantitative PCR were employed to detect the expression of SDF-1α protein and mRNA in retinal tissues respectively in injury group (n=28), sham operated group (n=28) and normal control group (n=12). Results The expression of SDF-1α protein and mRNA in retinas was higher than that in sham operated group and normal control group at different time points after injury (P<0.01), and it reached the peak at the 5th day after injury. The expression of SDF-1α protein and mRNA maintained a high level at the 14th day after injury. Conclusion The expression of SDF-1α protein and mRNA is up-regulated after partial optic nerve injury, and may last for a long time.

Objective To test the effect of bone marrow mesenehymal stem cells (MSCs) transplantation on oxidative stress and the development of pulmonary emphysema in rats. Methods SD rats (n=26) were randomly divided into three groups:normal control group (group A, n=8),emphysema group (group B, n=8) and emphysema+MSCs transplantation group (group C, n=10).Rat models of emphysema was established by exposing rats to cigarette smoking for 14 weeks. Then rats of group C received MSCs transplantation. At the 14th and 28th days after 4 course of MSCs transplantations, one rat in group C was sacrificed at each time point and their lungs were preserved in frozen sections. Survival of MSCs in the lung tissues were observed by fluorescence microscopy. Eight weeks after transplantations, lung sections were stained by hematoxylin and eo?sin (HE) to observe the morphological alterations.Mean linear intercept (MLI) and mean alveolar numbers (MAN) were also measured. Serum and lung malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity were also examined. Re?sults At the 14th day and 28th day after transplantations of MSCs, MSCs successfully localized to lung and survived in rat models of emphysema. Emphysematous changes of lung tissues were observed in both group B and group C. MLI was higher while MAN was lower in group B and C than those in group A (P<0.05). MLI and MDA levels in serum and lung were high?er while MAN level and SOD activity were lower in group B than those in group C (P<0.05).MDA levels in serum and lung was higher while SOD activity was lower in group B and C than those in group A (P<0.05). Conclusion MSCs transplanta?tions can effectively alleviates pulmonary emphysema in rat models which might through reducing oxidative stress .

Objective To analysis the human T lymphotropic virus (HTLV) infection status in Wenzhou among voluntaty blood donors.Methods Selected 72 417 voluntary blood donors of Wenzhou from from March,1,2016,to November,30,2016,to screen HTLV-Ⅰ / Ⅱ antibody by ELISA method.The positive samples were reexamined two times,two test results of samples were determined positive by ELISA.HTLV positive samples was confiemed by Western Blotting (WB).Results Screened 23 cases of anti-HTLV positive by ELISA method,then confirmed 9 cases of HTLV positive by Western Blotting (WB).HTLV infection rate of Wenzhou blood donors was 0.01％ (9/72 417).Conclusions HTLV infection was found among volunteer blood donors in Wenzhou,but the HTLV infection rate of volunteer blood donors in Wenzhou is still at a relatively low level.

Objective To investigate the feasibility and role of single-dose three-dimensional dynamic contrast-enhanced magnetic resonance angiography(SD 3D DCEMRA)in evaluating soft-tissue hemangioma of limbs.Methods The transit time(TT),signal intensity of peak enhancement(SPE)and duration of peak enhancement(DPE)of the femoral,popliteal and anterior tibial arteries at the level of the middle section were assessed in 30 healthy volunteers after intravenous bolus injection of single-dose contrast media at the rate of 3 ml/s.Forty-five patients with soft-tissue hemangioma and 9 patients with schwannoma of the extremities underwent both conventional MRI and SD 3D DCEMRA.The acquisition time of SD 3D DCEMRA ranged from 10 to 12 s,and early arterial,late arterial and venous phases of SD 3D DCEMRA images were acquired consecutively.The conventional MRI and SD 3D DCEMRA findings of the 45 patients with hemangioma were observed and compared with those of the 9 cases of sehwannoma.Results(1)The TT,SPE and DPE of the femoral,poplitcal and anterior tibial arteries were(15?5)s,(400?50),(11.9? 2.6)s;(19?7)s,(320?45),(16.8?3.6)s and(27?10)s,(270?39),(22.0?6.6)s respectively. The comparison of TT(F=6.91,P0.01).The tumoral feeding artery was visualized in all cases(100%).(3)Two cases of hemangioma missed on conventional MRI were correctly diagnosed on SD 3D DCEMRA owing to the visualization of both the tumoral mass and the feeding artery.(4)For schwannoma,neither the dynamic visualization of tumoral mass nor the feeding artery was demonstrated on SD 3D DCEMRA.Conclusion SD 3D DCEMRA is technically feasible to evaluate the limb soft-tissue hemangioma.Dynamic visualization of tumoral mass and demonstration of the tumoral feeding artery are the characteristic features of the tumor on SD 3D DCEMRA.

OBJECTIVE: To observe the effect of herbal-cake-partitioned moxibustion (HCPM) of "Shenque" (CV8) and "Daheng" (SP15) on abdominal pain, plasma β-endorphin (β-EP), uterine prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) levels, as well as splenetic natural killer cell (NK cell) activity in primary dysmenorrhea (PD) rats, so as to explore the specificity of acupoint function and the underlying mechanisms of moxibustion in relieving dysmenorrhea. METHODS: A total of 40 female rats were randomized into blank control, model, CV8-direct moxibustion, CV8-HCPM and SP15-HCPM groups (n＝8 rats in each). The PD model was established by subcutaneous injection of estradiol benzoate injection (0.2-0.5 mg/rat) for 10 consecutive days and intraperitoneal injection of oxytocin (2 U) 24 h after the last subcutaneous injection. Moxibustion or herbal-cake (composed of Radix Angelicae Sinensis, Rhizoma Chuanxiong, Radix Paeoniae Rubra, Cortex Cinnamomi, etc.)-partitioned moxibustion was applied to CV8, SP15 or umbilicus respectively for 7 moxa-cones every time, once daily for 10 successive days. The rats of the control and model groups were also restrained as those in the moxibustion groups. The writhing times within 30 minutes was recorded and the contents of plasma β-EP, uterine PGE2 and PGF2α were detected by ELISA, and NK cell activity was detected using MTT. RESULTS: Compared with the control group, the writhing times and the content of PGF2α in the uterus tissue were significantly increased in the model group (P<0.01), while the contents of plasma β-EP, uterine PGE2 and splenetic NK cell activity were significantly decreased (P<0.01). In comparison with the model group, the writhing times and uterine PGF2α content were obviously down-regulated in the SP15-HCPM, CV8-direct moxibustion and CV8-HCPM groups (P<0.05, P<0.01), and the contents of plasma β-EP and uterine PGE2, and splenetic NK cell activity were significantly increased (P<0.05, P<0.01). The therapeutic effects of CV8-HCPM group were significantly superior to those of SP15-HCPM and CV8-direct moxibustion groups in lowering writhing times and PGF2α level, and in up-regulating β-EP, PGE2 (P<0.05, P<0.01). The NK cell activity of CV8-HCPM group was significantly increased compared with that in the SP15-HCPM group(P<0.05). No significant differences were found between the SP15-HCPM and CV8-direct moxibustion groups in the levels of writhing times, plasma β-EP and uterine PGE2, PGF2α contents and splenetic NK cell activity (P>0.05). CONCLUSION: Moxibustion of both CV8 and SP15 can relieve abdominal pain in PD rats, which may be closely associated with its effect in suppressing PD-induced decrease of plasma β-EP and uterine PGE2 levels and splenetic NK cell activity and increase of uterine PGF2α. The therapeutic effect of CV8-HCPM is obviously better than that of SP15-HCPM and CV8-direct moxibustion.

OBJECTIVE: To explore the underlying mechanism of acupoint catgut embedding in improving primary dysme-norrhea (PD) in rats based on functional activities of the neuro-endocrine-immune (NEI) network. METHODS: Forty female rats were equally randomized into blank control, PD model, medication, and acupoint catgut embedding groups. The PD model was established by subcutaneous injection of estradiol benzoate (0.5 mg/rat on the 1st and 10th d, and 0.2 mg/rat from 2nd to 9th d) and oxytocin (2 U/rat, i．p.). Rats of the medication group were treated by intragastric perfusion of fenbid (0.8 mL/rat, 125 mg/100 mL), once daily for 10 days. The catgut embedding was applied to bilateral "Ciliao" (BL 32), "Sanyinjiao" (SP 6) and "Guanyuan" (CV 4) before modeling. The body writhing times in 30 minutes were recorded, plasma β-endorphin(β-EP) content, and prostaglandin E 2 (PGE2) and prostaglandin F 2 α (PGF2α) contents in the uterus tissue were assayed using ELISA, and the activity of natural killer cell (NK cell) in the spleen tissue was detected using 3-(4，5-dimethyl-2-thiazolyl)-2，5-diphenyl-2-H-tetrazolium bromide (MTT) method after isolation and co-culture with K 562 cells. RESULTS: The body writhing times were no-tably more in the model group than in the control group (P<0.01), and obviously fewer in both medication and catgut embedding groups than in the model group (P<0.01). After modeling, the plasma β-EP and uterus PGE2 contents and splenic NK cell activity were significantly decreased (P<0.01), while the uterus PGF2α content was evidently increased in the model group relevant to the control group (P<0.01). Following the treatment, plasma β-EP and uterus PGE2 contents and splenic NK cell activity were considerably up-regulated (P<0.01), and uterus PGF2α content was markedly down-regulated in both medication and acupoint catgut embedding groups (P<0.01), suggesting an involvement of the NEI network in catgut embedding-induced improvement of PD. The therapeutic effect of catgut embedment was markedly superior to that of medication in up-regulating splenic NK cell activity (P<0.01). No significant differences were found between the medication and catgut embedding groups in the body writhing times within 30 min, and in the levels of plasma β-EP and uterus PGE2 and PGF2α (P>0.05). CONCLUSION: The acupoint catgut embedding has a significant efficacy in relieving PD in rats, which may be related to its effect in up-regulating plasma β-EP, uterus PGE2 contents and splenic NK cell activity and in down-regulating uterus PGF2α level.

OBJECTIVETo probe into the best acupoint-prescription for the simple acupoint catgut embedding therapy in treatment of bronchial asthma.

METHODSForty-eight Wistar rats were randomly divided into six groups: the simple acupoint catgut embedding at Feishu (BL 13), Danzhong (CV 17) and Shenshu (BL 23) group (group A), the simple acupoint catgut embedding at Shenshu (BL 23) group (group B), the simple acupoint catgut embedding at Feishu (BL 13) and Danzhong (CV 17) group (group C), Dexamethasone group (group D), model group (group E), and control group (group F), 8 rats in each group. The asthmatic models were established by Ovalbumin (OVA) except group F. Rats in group A, B and C were treated with catgut embedding at the corresponding acupoints from the first experimental day. In group E and D, Dexamethasone and sterile were intraperitoneal injected respectively from the 15th experimental day, once a day for 2 weeks consistantly. No interventions were added on group F. For the six groups, the symptoms of asthmatic attack were observed and the pathologic changes of lung tissue were examined.

RESULTS(1) The times of sneeze and rhinocnesmus scratching nose in group E were increased significantly compared with those in group F (both P < 0.01), but there was no significant difference between group D and F (both P > 0.05). As compared with that in group E, except the times of sneeze in group C, the times mentioned above were decreased significantly in all the treatment groups (all P < 0.01). There was no significant difference between group A and D (both P > 0.05), but the times mentioned above were increased significantly in group B and C as compared with that in group A (all P < 0.01). (2) The symptoms of asthmatic attack and the pathologic changes of airway tissue were all alleviated in group A, B and C, but a better amelioration was observed in group A, with no mucus epistom in the bronchial lumen and few infiltrations of inflammatory cells around the bronchi.

CONCLUSIONThe improvement of the simple acupoint catgut embedding at "Feishu" (BL 13), "Danzhong" (CV 17)and "Shenshu" (BL 23) on the airway inflammation in asthmatic rats is better than that of catgut embedding at "Feishu" (BL 13) and "Danzhong" (CV 17) or at "Shenshu" (BL 23) only.

Acupuncture Points ; Acupuncture Therapy ; methods ; Animals ; Asthma ; pathology ; therapy ; Catgut ; Male ; Medicine, Chinese Traditional ; Random Allocation ; Rats ; Rats, Wistar

Objective:To use polyamidoamine(PAMAM)dendrimer as gene delivery system for survivin gene anti- sense oligodeoxynucleotide(asODN)transfection for inhibition of HepG2 cancer cell growth.Methods:The first to the fifth generation of PAMAM and asODN were used to prepare a complex:PAMAM-asODN.The morphology of PAMAM- asODN was observed using agrose electrophoresis and atomic force microscope(AFM).PAMAM-asODN was then used to transfect HepG2 cells and cells transfected with asODN served as control.The transfection efficacy of PAMAM-asODN into HepG2 cells was observed under confocol microscope,the surviving mRNA expression was analyzed by RT-PCR,and the inhibition of HepG2 cell growth was determined by MTT assay.Results:Agrose electrophoresis showed strong complexing action between PAMAM and asODN and they formed a complex with a diameter of 25 nm.Confocol microscope showed the transfection efficacy of PAMAM-asODN was higher than that of asODN.RT-PCR showed a decreased expression of sur- vivin mRNA in PAMAM-asODN transfected cells.MTF results demonstrated that the growth of HepG2 cell was obviously inhibited after transfection of PAMAM-asODN and the inhibition rate increased with culture time,concentration of com- plex,the generation of PAMAM.PAMAM-asODN at 6.0?mol/L G4.0 resulted in a 55% inhibition of HepG2 cells 96 h after culture.Conclusion:PAMAM dendrimers can efficiently mediate the entry of survivin asODN into HepG2 cells,re- sulting in inhibition of HepG2 cells.PAMAM might be a promising gene carrier for potential molecular therapy of cancer.

OBJECTIVETo observe the effect of Cordyceps sinensis (CS) powder on renal oxidative stress and mitochondria functions in 5/6 nephrectomized rats, and to primarily explore its possible mechanisms.

METHODSTotally 30 male Sprague-Dawley rats were divided into the sham-operation group, the model group, and the treatment group by random digit table, 10 in each group. A chronic kidney disease (CKD) rat model was prepared by one step 5/6 nephrectomy. Rats in the treatment group were intragastrically administered with CS powder solution at the daily dose of 2 g/kg, once per day. Equal volume of double distilled water was intragastrically administered to rats in the sham-operation group and the model group. All medication lasted for 12 weeks. The general condition of rats, their body weight, blood pressure, 24 h proteinuria, urinary N-acetyl-β-D-glucosaminidase (NAG), serum creatinine (SCr) , and blood urea nitrogen (BUN) were assessed before surgery, at week 2, 4, 6, 8, 10, and 10 after surgery. Pathological changes of renal tissues were observed under light microscope. Morphological changes of mitochondria in renal tubular epithelial cells were observed under transmission electron microscope. Activities of antioxidant enzymes including reduced glutathione (GSH), manganese superoxide dismutase (MnSOD), and malondialdehyde (MDA) in fresh renal tissue homogenate were detected. Mitochondria of renal tissues were extracted to detect levels of mitochondrial membrane potential and changes of reactive oxygen species (ROS). And expressions of cytochrome-C (Cyto-C) and prohibitin in both mitochondria and cytoplasm of the renal cortex were also measured by Western blot.

RESULTS(1) Compared with the sham-operation group, body weight was significantly decreased at week 2 (P <0. 01), but blood pressure increased at week 4 (P <0. 05) in the model group. Compared with the model group, body weight was significantly increased at week 12 (P <0. 01), but blood pressure decreased at week 8 (P < 0. 01) in the treatment group. (2) Compared with the sham-operation group, 24 h proteinuria, urinary NAG, blood SCr and BUN significantly increased in the model group (all P <0. 01). Compared with the model group, blood and urinary biochemical indices all significantly decreased in the treatment group (all P <0. 01). (3) Results of pathological renal scoring: Glomerular sclerosis index, scoring for tubulointerstitial fibrosis, degree of tubulointerstitial inflammatory infiltration were all obviously higher in the model group than in the sham-operation group (all P <0. 01). All the aforesaid indices were more obviously improved in the treatment group than in the model group (all P <0. 01). (4) Compared with the sham-operation group, activities of MnSOD and GSH-Px were significantly reduced, but MDA contents obviously increased in the renal cortex of the model group (all P <0. 01). Compared with the model group, activities of MnSOD and GSH-Px obviously increased (P <0. 05, P <0. 01), but MDA contents obviously decreased in the renal cortex of the treatment group (P <0. 01). (5) Compared with the sham-operation group, the mitochondrial membrane potential significantly decreased, but ROS levels significantly increased in the model group (all P <0.01). Compared with the model group, mitochondrial transmembrane potential increased in the treatment group, thereby inhibiting the tendency of increased production of ROS (both P < 0. 01). (6) Results of Western blot showed that, compared with the sham-operation group, expression levels of mitochondrial Cyto-C and Prohibitin were significantly reduced in the renal cortex (P <0. 01), but significantly elevated in the cytoplasm of the model group (P <0. 01). Compared with the model group, each index was obviously improved in the treatment group with statistical difference (P <0. 05, P <0. 01).

CONCLUSIONCS powder had renal protection, and its mechanism might partially depend on in- hibition of oxidative stress and protection for mitochondria.

Acetylglucosaminidase ; metabolism ; Animals ; Blood Urea Nitrogen ; Cordyceps ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; Kidney Cortex ; Kidney Diseases ; Kidney Function Tests ; Male ; Malondialdehyde ; metabolism ; Mitochondria ; Nephrectomy ; Oxidative Stress ; drug effects ; Proteinuria ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism