BACKGROUND: The commonly used culture methods for primary intervertebral disc cells are type Ⅱcol agenase alone digestion method, and type Ⅱ col agenase combined trypin digestion method. However, the acquired cells are few. OBJECTIVE: To acquire nucleus pulposus cells from human degenerative intervertebral disc using a systemic and simplified method. METHODS: Nucleus pulposus cells were isolated and cultured. The morphology of nucleus pulposus cells was observed under inverted phase contrast microscope every day. Primary and subcultured cellsuspension was applied for the determination of cel viability using trypan blue staining. The cel growth was detected with MTT assay. The cel morphology was observed under laser confocal microscopy. RESULTS AND CONCLUSION: Nucleus pulposus cells were successful y isolated from from human degenerative intervertebral disc using the improved method, and cells were subcultured to passage 3. Primary cells were fusiform shaped, while cells at passage 1 and 2 were triangle or polygonal, which were similar to fibroblasts. When cells almost reached the confluence, the cells showed slabstone-like appearance. Trypan blue staining showed that, the primary cel viability was 99%, and passage 3 cells had 93%-95% viability. The proliferation of nucleus pulposus cells gradual y decreased as the generation increased. Compared with passage 1 cells, passage 2 and 3 cells at logarithmic phase trended to be smoother. The cel morphology observed under laser confocal microscopy was similar to the results under phase contrast microscope. The improved simple method can successful y acquire a variety of cells from human degenerative intervertebral disc, and these cells show fine biological properties.

BACKGROUND: Ascariasis is one of the most common human parasitic infections worldwide. In some rare cases, ascariasis may cause serious consequences even sudden death. This study was undertaken to review the life-threatening complications of ascariasis in trauma patients reported in the literature. DATA SOURCES: Relevant articles about ascariasis and trauma were searched from Pubmed, Google scholar, Scirus, and Wanfang databases. RESULTS: Twenty-four patients with ascariasis were collected from 21 articles searched. Most of these patients were from tropical and subtropical countries. Of the 24 patients, 12 were children. Their major complications occurred in the airway passage and digestive tract. There were 3 fatal cases in these patients. Twelve of the 24 patients described in 10 articles were reported in the last 10 years. CONCLUSIONS: Early diagnosis and prompt intervention are essential to minimize the high morbidity and mortality of these serious complications in trauma patients. Physicians should be aware of the possibility of Ascaris infection in a trauma patient from endemic area of ascariasis. History of Ascaris infection and routine examination of feces forAscaris eggs may be helpful to make a correct diagnosis.

Objective To investigate the therapeutic effects of latarjet nerve and pylous preserved pancreaticoduo-denectomy (LPPPD). Methods Clinical data and postoperative follow-up of Latarjet nerve and Pylous Preserved Pancreaticoduodenectomy since1996 of 32 cases were analyzed retrospectively, and 36 cases being carried out con-temporaneous Pylous Preserved Pancreaticeduodenectomy(PPPD) were compared with. Results The recovery time of postoperative gastrointestinal function recory time is five days of LPPPD group on average; but the time is eight days of PPPD group on average, and significantly slower than LPPPD group ( t = 3.01, P < 0.05 ) ; the occurrence of abdominal distenal, retention of gastric juice and enterogastric recurrent flow are significantly slower in LPPPD group than that in PPPD group( P < 0.05 ). Conclusion The postoperative gastrointestinal function recovered fas-ter, and the postoperative complications were less in LPPPD group than that in PPPD group.

Four screening methods, colorimetric assay, blood-plate hemolysis method, surface tension activ- ity and oil spreading technique were introduced to isolate strains producing bio-demulsifiers from 6 different bacteria source samples. The results of various screening methods were evaluated in this paper. Seventeen demulsifying strains were obtained, which are qualified in demulsification test of kerosene model emulsions. Among them, 5 strains showed high demulsifying ability, achieving 70% plus demulsifying ratio within 24 hours. Petroleum-contaminated soil, excess sludge from biological process treating oilfield produced water and sludge from municipal wastewater treatment plant were the best among all tested sources. Due to the determination limit, the colorimetric assay and blood-plate hemolysis method are not competent to screen bio-demulsifiers strains. The measurement of surface tension and oil spreading method were easy but accu- rate methods to isolate bio-demulsifiers strains. Although demulsification test of model emulsion is an effec- tive technique to target strains with the capability of breaking emulsions, it is sophisticated and time-con- suming. Thus it is recommended to use surface tension and oil spreading methods in pre-screening and vali- date the results in demulsification test with kerosene model emulsions.

Objective To discuss the short-term clinical effect including functional change of lipiodol- arsenic trioxide emulsion on the primary hepatic carcinoma.Methods Fifty-two patients undergone arterial chemoemblization were selected and then randomly divided into two groups:treatment group(n=27)and control group(n=25).Patients in treatment group were treated with lipiodol-arsenic trioxide,while those in control group treated with mitomycin,epirubicin,cisplatin or lipiodol.Clinical symptoms and six liver function parameters were observed and analized.Results The clinical symptoms of patients in treatment group improved much better than those in control group,and the liver function impairment of patients in treatment group also decreased more than those in control group.Conclusions Lipiodol-arsenic trioxide is an effective and safe intervention-therapeutic embolization material for primary hepatic carcinoma.

Objective To study the effect of different concentration of 1,25-dihydroxyvitamin D3[1,25(OH)2D3] on cell proliferation,differentiation and the expression of vitamin D receptor (VDR) in mouse MC3T3E1 osteoblast.Methods Osteoblast were cultured in medium with different concentrations of 1,25(OH)2D3.Incubated for 48 h,cell proliferation of osteoblast were examined by MTT reduction assay (mono-nuclear cell direc cytotoxicity assay),the osteocalcin (OC) levels in cell medium were detected by ELISA,and the expression of VDR mRNA and protein were examined by using SYBR Green real-time PCR and Western blot,respectively.Results 1.After incubation with 1,25(OH)2D3 for 48 h,the number of MC3T3E1 osteoblast was significantly less than that in control group(P0.05).3.SYBR Green real-time PCR and Western blot results showed that the expression of VDR mRNA as well as VDR protein of osteoblast in 10-8,10-9 mol/L experimental groups were significantly higher than those in control group (Pa0.05).Conclusions Cell proliferation of mouse osteoblast can be inhibited,while the cell differentiation was promoted by 1,25(OH)2D3.1,25(OH)2D3 up-regulated the expression of VDR in mouse osteoblast,which suggested that the VDR signal pathway may play some role in proliferation and differentiation of osteoblast.

ObjectiveTo observe the effect of Jiuqiang Naoliqing (JNQ) on the behavior of Kunming mice.MethodsSpontaneous movement, Morris Water Maze, Rotarod, anti caffeine test, sleeping time of pentobarbital sodium, subthreshold dose of pentobarbital sodium, and anti pentylene tetrazol test were adopted to evaluate the behavioral changes. ResultsCompared with the control group, the low dose of JNQ can increase spontaneous movement of the mice, the middle and high dose of JNQ can increase time on the rotating rods. JNQ can also increase sleeping time of pentobarbital sodium test and percent of falling asleep in subthreshold dose of pentobarbital sodium test, as well as antagonize caffeine's effect on mice. ConclusionJNQ can also do sedative and hypnotic effect on Kunming mice as well as improve their ability of balance and coordination.

The genus Xestospongia is one of the most widespread genera of sponges, containing abundant secondary metatolites with novel structures and potent bioactivities. The main structure types of secondary metatolites found in this genus are alkaloids, quinines, terpens, steroids, lipids, polyketones, etc. These metatolites exhibit a variety of bioactivities, such as cytotoxic, antibacterial and antiviral activities. This paper reviews the progress in the chemistry and pharmacological activities of the second metabolities from sponges of Xestospongia, especially for recent five years, with the aim for further research.
Animals ; Secondary Metabolism ; Xestospongia ; chemistry

OBJECTIVETo investigate the expression of TSC1, TSC2, p-mTOR, p-4E-BP1, p-p70S6K and p-S6 in refractory epilepsy associated malformation of cortical development (MCD) tissues.

METHODSA total of 43 cases of refractory epilepsy were involved in the study, and all the patients were treated in Xuanwu Hospital during 2005 - 2008, including focal cortical dysplasia type IIa (11 cases) and type IIb (11 cases), tuberous sclerosis complex (10 cases) and ganalioglioma (11 cases), and other 12 cases were used as control. These cases were divided into 7 study groups and immunohistochemical EnVision method was used. To detect the location and intensity of TSC1, TSC2, p-mTOR, p-4E-BP1, p-p70S6K and p-S6 expression in every group. Then the Image-Pro Plus 6.0 image processing and analysis software were used to measure the number, area, integrating absorbance (IA) of positive cells in every samples. The statistical software SPSS 16.0 was used to analyze the data.

RESULTSThe immunolocalization of TSC1 and TSC2 was similar. It could be observed the expression of various levels in the cytoplasm of dysmorphic neurons, balloon cells, giant cells, ganglioglioma cells and normal neurons. TSC1 staining in normal neurons was more notably than others but TSC2 staining in giant cells was weaker than other samples. p-mTOR mainly presented in giant cells, which could also be observed in astrocyte. P-4E-BP1 presented in the cytoplasm and nuclear membrane of balloon cells, giant cells and ganglioglioma cells, the staining of giant cells was stronger than balloon cells, but their staining were weaker than ganglioglioma cells. P-p70S6K mainly expressed in giant cells and less commonly presented in balloon cells. P-S6 typically presented in all abnormal glioneuronal cells and it nearly did not present in the normal neurons of N-CTX group.

CONCLUSIONSPI3K pathway, at least in part, involves in the occurrence of MCD, and may play an important role in the pathogenesis.

Adaptor Proteins, Signal Transducing ; metabolism ; Adolescent ; Adult ; Child ; Epilepsy ; metabolism ; pathology ; Female ; Ganglioglioma ; metabolism ; pathology ; Humans ; Male ; Malformations of Cortical Development ; metabolism ; pathology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphoproteins ; metabolism ; Ribosomal Protein S6 Kinases ; metabolism ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; Signal Transduction ; TOR Serine-Threonine Kinases ; metabolism ; Tuberous Sclerosis ; metabolism ; pathology ; Tumor Suppressor Proteins ; metabolism ; Young Adult

OBJECTIVETo observe the preventive effect different compatibilities of Ramulus Cinnamomi (RC) and peony in Guizhi Decoction (GD) on diabetic cardiac autonomic neuropathy (DCAN).

METHODSTotally 60 male rats were randomly divided into 5 groups, i.e., the blank control DM group, the model group, the methycobal group, the 1:1 (RC/peony) Guishao group, the 2:1 Guishao group, and the 1:2 Guishao group, 10 in each group. Rats were pretreated with corresponding drugs for 1 week, and then induced diabetes by intraperitoneal injection of STZ. Drugs were administrated by gastrogavage for 4 more weeks after STZ-injection. Enzyme-linked immunosorbent assay (ELISA) was employed to detect levels of tyrosine hydroxylase (TH), choline acetyltransferase (CHAT), nerve growth factor. (NGF), and ciliary neurotrophic factor (CNTF) in myocardial homogenates.

RESULTSAfter 4-week modeling, body weight (BW) was obviously lower, but blood glucose (BG) was higher in STZ rats than in rats of the blank control DM group. There was no statistical difference in BW or BG among the 5 groups (P >0.05). Compared with the blank control group, TH, TH/CHAT, and NGF in left ventricle and ventricular septum increased, CHAT and CNTF increased in the model group (P < 0.05, P < 0.01). Compared with the model group, TH and TH/CHAT in left ventricle decreased (P < 0.05, P < 0.01), CNTF in left ventricle increased (P < 0.05), CHAT in left ventricle and ventricular septum increased (P < 0.05, P < 0.01) in the methycobal group. TH and TH/CHAT in left ventricle and ventricular septum decreased, CNTF in left ventricle and ventricular septum increased (P < 0.05, P < 0.01), CHAT in left ventricle and ventricular septum increased (P < 0.01), NGF in ventricular septum decreased (P < 0.01) in the 1:1 Guishao group. TH/CHAT in left ventricle decreased (P < 0.01), CHAT and CNTF in left ventricle and ventricular septum increased (P < 0.05, P < 0.01) in the 1:2 Guishao group. Compared with the methycobal group, CHAT in left ventricle decreased, TH and TH/CHAT in left ventricle increased in the 2:1 Guishao group (P < 0.05, P < 0.01). TH and TH/CHAT in ventricular septum decreased (P < 0.05), CHAT and CNTF in left ventricle and ventricular septum increased (P < 0.05, P < 0.01) in the 1:1 Guishao group. Compared with the 1:2 Guishao group and the 2:1 Guishao group, CHAT in left ventricle increased, TH/CHAT in left ventricle decreased, TH and TH/CHAT in ventricular septum decreased, CHAT in ventricular septum increased, CNTF in left ventricle and ventricular septum also increased in the 1:1 Guishao group (all P < 0.01).

CONCLUSIONSSTZ model rats had autonomic neural injury, manifested as lowered vagal nerve activity and hyperactive sympathetic nerves. GD could effectively suppress hyperactive cardiac sympathetic nerves and protect the vagus. Besides, GD (1:1) showed the optimal effect in regulating the balance of cardiac autonomic nerves and could be used in early prevention of DCAN.

Animals ; Blood Glucose ; Choline O-Acetyltransferase ; Diabetic Neuropathies ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Heart ; Heart Ventricles ; Male ; Myocardium ; Nerve Growth Factor ; Paeonia ; Rats ; Tyrosine 3-Monooxygenase