Objective:To observe the effect of Gegen Qinlian decoction on high sensitive C-reactive protein(hs-CRP)and interleu-kin-6(IL-6)in serum of the rats with periodontitis and the IL-6 expression level in periodontal tissues.Methods:45 Wistar rats were randomly divided into 2 groups:control group N(n =1 0),periodontitis model group P(n =35).After periodontitis model was identi-fied by the examination of 5 rats,the rest rats were randomly divided into 3 groups(n =1 0)and treated by gavage of saline(group P0 ), metronidazole(group P1 )and Gegen Qinlian decoction(group P2 )respectively for 4 weeks.Then all the rats were sacrificed.Serum was immediately harvested for the test of serum hs-CRP and IL-6 levels by ELISA.Attachment loss level(AL)was examined by pa-thology.IL-6 expression in periodontal tissues was examined by S-P immunohistochemistry.Results:The inflammation of periodontal tissues in P1 ,P2 were improved more than that in P0 .AL in group P2 were lower than that in other groups(P <0.05).The IL-6 expres-sion in periodontal tissues and the levels of serum hs-CRP and IL-6 of group P2 were lower than those of other groups(P <0.05).Ser-um hs-CRP level was positively correlated with IL-6 level.Conclusion:Gegen Qinlian decoction may inhibit the development of peri-odontitis by depressing the expression of serum hs-CRP and IL-6.

Objective:To observe the effect of Gegen Qinlian decoction on serum high sensitive C-reactive protein(hs-CRP)expres-sion of rats with periodontitis and atherosclerosis(P-AS).Methods:40 male Wistar rats were randomly divided into 2 groups:control group(A,n=1 0),P-AS group(B,n=30).5 rats in group A and B were used for the identification of P-AS model.Then rats with P-AS in group B were randomly divided into 5 groups(n=5)and administered with saline(B1 ),atorvastatin(B2),metronidazole (B3),atorvastatin+metronidazole(B4)and Gegen Qinlian decoction(B5)respectively for 4 weeks.Serum hs-CRP level was assayed by ELISA.Periodontal attachment loss(AL)and artery change were examined by pathology.Results:Serum hs-CRP level in group B1-5 was higher than that of group A(P<0.01 ).Serum hs-CRP level of group B4 and B5 was lower than that of group B1-3(P<0.01 ),B4 vs B5,P>0.05.The inflammation of periodontal tissues in group B2-5 was improved more than that in group B1 .AL in group B4 and B5 were lower than that in other groups(P<0.01 ),B4 vs B5,P>0.05.Histopathological observation of arteries re-vealed that there was no foam cell in group B4 and B5 ,the artery wall in group B4 and B5 was more even and muscle fibers were ar-ranged more regular.Conclusion:Gegen Qinlian decoction may decreas serum hs-CRP expression and depress the development of pe-riodontitis and atherosclerosis.

In order to investigate the biological effects of the VP22?-mE6?/mE7 built-in adjuvant fusion protein vaccine on the tumor associated with HPV-16 infection.HSV-1 VP22? and HPV-16 mE6?/mE7 genes were cloned,and the pET28a-VP22?-mE6?/mE7 recombinat prokaryotic expression vector was constructed.Vector was transformed into Rosetta(DE3)E.coli string and expressed under the induction of IPTG.The re-naturalized protein was then purified via Ni2+ affinity adsorbent column and identified by SDS-PAGE and Western blot.Purified protein was immunized BalB/C and C57BL/6 mice to evaluate the immunogenicity and anti-tumor activity.The expressed recombinant protein formed as inclusion body with a prediction MW about 34kDa and contained approximately 45% of total somatic protein.The VP22?-mE6?/mE7 immunization induced higher titer of specific IgG against HPV,higher level of lymphocyte proliferation and better effect on suppressing HPV16 positive TC-1 tumor growth than the mice immunized with mE6?/mE7 alone.The results showed that the recombined built-in adjuvant vaccine could induce specific cellular immune response in vitro and inhibit the TC-1 tumor proliferation in vivo,that would be a foundation for further studies and developments of inner adjuvant vaccines.

AIM To observe the oxidant stress and opoptotic effects of anisodine hydromide (AH) on chronic cerebral hypoperfusion (CCH) rats.METHODS In vivo CCH models were established in adult male SpragueDawley rats by permanent ligation of bilateral common carotid arteries [two-vessel occlusion (2-VO)] surgery.Rats were randomly divided into six groups,sham group,model group,positive group of n-butylphthalide and sodium chloride injection,and AH groups (1.2 mg/kg high-dose group,0.6 mg/kg medium-dose group,and 0.3 mg/kg low-dose group).Antioxidant indices including the activity of SOD,CAT,LDH and iNOS and the content of GSH and NO were measured.In the in vitro trial,PC12 cells were divided into control group,model group,positive group of n-butylphthalide,and AH groups (100 μmol/L high-dose group,50 μmol/L mediumdose group,and 25 μmol/L low-dose group),and the hypoxic models were established by treating PC12 cells with CoCl2.The cells had their release of NO and LDH detected,their cellular apoptosis determined by Hochest 33342 fluorescence staining,and the expression of P53 protein identified by IF (immunofluorescence) and Western blotting method.RESULTS The in vivo trial revealed AH's enhancement in serum SOD activity and inhibition in serum iNOS activityof the CCH rats,and its power in the cerebral GSH and LDH release reduction.The in vitro trial showed the resultant lower LDH and NO release,decreased number of neuro-apoptosis,and inhibited P53 pro tein expression after AH intervention.CONCLUSION The antioxidant and antiapoptotic effects of AH on CCH rats may be associated with down regulation of P53 protein.

An autopsy case of sudden death induced by alimentary tract hemorrhage was presented, which was caused by the unexpected rupture of clinically unrecognized tuberculous abdominal aortic aneurysm (TAAA). The initial diagnosis was made of the syndrome of coronary heart disease and hypertensive disease. The detailed autopsy showed that the alimentary tract hemorrhage was caused by a sudden rupture of the mass after posture changing was ascertained as the cause of death. The diagnosis of TAAA was determined by the autopsy findings. Analysis for the medical dispute of TAAA was described, and the difficulty of the diagnosis and medico-legal implications were also discussed.
Aneurysm, Ruptured/diagnosis* ; Aortic Aneurysm, Abdominal/diagnosis* ; Autopsy ; Death, Sudden ; Hemorrhage/etiology* ; Humans ; Tuberculosis/diagnosis*

OBJECTIVETo investigate the protective effect of neferine against damages of endothelial cells induced by lysophos-phatidylcholine (LPC) and the relationship with asymmetric dimethylarginine (ADMA).

METHODThe human umbilical vein endothelial cells (HUVEC-12) were treated with LPC (10 mg x L(-1)) for 24 h to establish the model of endothelial cells damages; HUVECs were prior exposed to neferine (0.1, 1.0 or 10.0 micromol x L(-1) ) for 1 h, and then exposed to LPC in the presence of the neferine for 24 h. At the end of the experiment, the cultured medium was collected for measuring the concentration of nitric oxide (NO), aleic dialdehyde (MDA) as well as ADMA and the cells were collected for measuring the level of intracellular reactive oxygen species (ROS).

RESULTCompared with control group, exposure of endothelial cells to LPC (10 mg x L(-1)) for 24 h significantly increased the concentration of MDA and ADMA in the medium and the level of intracellular ROS and coinstantaneously significantly decreased the concentration of NO in the medium. Neferine (0.1, 1.0 or 10.0 micromol x L(-1)) significantly inhibited the elevated concentration of MDA, ADMA as well as the level of intracellular ROS and coinstantaneously significantly attenuated the decreased level of NO induced by LPC.

CONCLUSIONNeferine can protect the endothelial cells against damages induced by LPC and the protective effect is related to the decrease of the concentration of ADMA.

Arginine ; analogs & derivatives ; metabolism ; Benzylisoquinolines ; pharmacology ; Cell Line ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Lysophosphatidylcholines ; pharmacology ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Reactive Oxygen Species ; metabolism

OBJECTIVETo investigate the role of neuronal apoptosis in organophosphorus poisoning-induced delayed neuropathy (OPIDN) and its dynamic pathological changes.

METHODSTo establish OPIDN animal model, triorthocresyl phosphate (TOCP)was given to hens with a single dose (1 000 mg/kg, im). Changes of neuropathology, number of neurons and apoptotic cells in the third lumbar spinal cord were observed by HE, Nissl and TUNEL methods 3, 5, 7, 10, 14, 18 days after injection.

RESULTSThe hens showed OPIDN typical signs (progressive ataxia and hypotonia) about 9 days after TOCP exposure. HE staining revealed dark red nucleus in neurons of anterior horn of lumbar spinal cord 5 days after exposure, but this phenomenon disappeared 18 days later. Nissl method showed that the number of neurons in anterior horn of spinal cord decreased [from (82 +/- 4) cell/mm(2) to (66 +/- 6) cell/mm(2)]. TUNEL positive cells began to appear [(22 +/- 2) cell/mm(2)] 5 days after TOCP exposure, and reached the peak [(27 +/- 3) cell/mm(2)] 7 days later, and disappeared 18 days later.

CONCLUSIONNeuronal apoptosis in anterior horn of spinal cord of hens appeared in OPIDN, suggesting that cellular apoptosis may play an important role in the pathogenesis of OPIDN.

Animals ; Apoptosis ; drug effects ; Chickens ; Female ; In Situ Nick-End Labeling ; Insecticides ; toxicity ; Models, Animal ; Neurons ; drug effects ; Spinal Cord ; drug effects ; Tritolyl Phosphates ; toxicity

OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.

Objective To evaluate the effect of artemether on the cell cycle and the radiosensitivity in human nasopharyngeal carcinoma cell line CNE-1.Methods Cell growth inhibition was assessed with MTT.The method of colony-forming was used to detect the radiation sensitivity.Cell cycle distribution was analyzed by using flow cytometry.The protein expressions of clyclin B1 and Weei were detected by using Western blot.Results The growth of CNE-1 cells was inhibited in a dose-dependent manner.The concentration of 20 μmol/L artemether had radiosensitive effect on CNE-1 cells at 24 h after administration,and SER was 1.481.When CNE-1 cell was irradiated,the G2/M cells increased (t =4.59,P ＜ 0.05).After exposure to combination of artemether and irradiation,the G2/M cells were decreased (t= 10.60,P ＜ 0.05).Western blot showed that artemether increased the level of cyclin B1 expression and inhibited the level of Weel expression.Conclusions The noncytotoxic concentration of artemether could enhance radiosensitization of CNE-1 cells.The radiosensitivity enhancement of artemether might depend on the exposure time.The effect is most obvious when radiation is delivered 24 h after expose to artemetherr.The radiosensitizing effect could be related to apoptosis.

Objective: To discuss the procedure and clinical effect of retroperitoneal laparoscopic nephropexy (RLN).Methods: From August 2001 to June 2006, RLN was performed on 28 female patients aged 26-45 years old (mean, 34±2.5) with symptomatic nephroptosis, including 15 with the right kidney, 12 with the left, and 1 with both. The preoperative complaint of patients included subjective symptoms (constant and recurring pain in 28 patients) and objective symptoms (upper urinary infections in 16, hematuria in 12, and upper tract obstruction in 12). One patient underwent nephropexy via the transperitoneal approach and the others underwent nephropexy via the retroperitoneal approach. A retroperitoneoscopic procedure was performed after positioning the patients in the flank position. Digital preparation of the retroperitoneal space was made and standardized trocar was placed. The key step of the surgery was complete exposure of the kidney within Gerota' fascia, which was aimed to separate the potential adhesions between the colon and kidney or between the inferior blood vessels of the kidney. Nephropexy was performed between the fibrous capsule at the lower pole of the kidney and the dissected psoas muscle, using three sutures placed by intracorporeal technique or the percutaneous needle both for introduction and removal of the suture; the sutures were separately tied over the sacrospinalis fascia. Results: The mean operative time was (125±9) min (ranging 115-240 min); the mean postoperative hospital stay was (9±1.2) days, largely owing to the required 5-12 days' bed rest. During a mean follow-up of (24±4.2) months(ranging 3 to 70 months), 3 patients had paresthesia, 5 had constant and recurrent ache, 20 were completely free of pain, and 4 had micro-hematuria. One patient had further episodes of pyelonephritis and upper tract obstruction after operation. Intravenous pyelogram(IVP) revealed that the ptosis incorporated into more than one vertebral body in 2 patients. Postoperative renal function test showed an improvement in renal function. Conclusion: RLN is mini-invasive and has less complication. The procedure should be considered as one of the optimal therapy for nephroptosis.