Child ; Humans ; Liver Failure ; therapy ; Liver Transplantation ; Male ; Plasma Exchange

Objective In order to explore the roles of TLR2 and TLR4 in the hepatocyte dam- age caused hy hepatitis B virus infection,and to find whether LPS can affect the damage of hepato- cytes pre-and pos-HBV infection,we detected the changes of TLR2 and TLR4 expressions in hu- man hepatocyte lines HepG2 cells and 2.2.15 cells.Methods HepG2 ceils are most similar to normal human hepatocytes and 2.2.15 ceils are HepG2 cells infected with HBV.We selected these two cell lines to study the differences of TLR2 and TLR4 expression between HepG2 cells before and after HBV infection.In this research,both HepG2 and 2.2.15 cells were stimulated with 0?g/ml, 1?g/ml,10?g/ml,100?g/ml,1 mg/ml and 10 mg/ml LPS.Then the expression of protein of TLR2 and TLR4 were examined by immuno-histochemistry(IHC).The cnRNA of HepG2 and 2.2.15 ceils stimulated with 0?g/ml and 10 mg/ml LPS were examined by reversal transcription-pol- ymerase chain reaction(RT-PCR).Thereafter,the apoptosis of HepG2 and HepG2.2.15 cells were examined by flow cytometry(FC),and the expressions of HBsAg and HBeAg of HepG2.2.15 cells tested with Abbott kits.Results IHC and RT-PCR analysis revealed that TLR2 and TLR4 expres- sions could he detected in both HepG2 and 2.2.15 cells.Moreover,without immune activation, TLR2 and TLR4 expressions were higher in the presence of higher concentrations of LPS.FC analy sis revealed that no apoptosis detected in HepG2 ceils stimulated with LPS in this research,but apop- tosis could be detected in 2.2.15 cells when treated with the same factors.Furthermore,the apoptosis ratios increased with the increase of LPS concentrations.When concentrations of LPS were 1?g/ml, 10?g/ml,100?g/ml,1 mg/ml and 10 mg/ml,the apoptosis ratios were 1.94%,3.03%,3.50%, 3.72%,5.30%,respectively.Abbott analysis revealed that expressions of HBsAg and HBeAg of 2.2.15 cells stimulated with LPS were lower than those not stimulated with LPS.Conclusion HBV can affect the expressions of TLR2 and TLR4 in HepG2 cell lines.LPS can lead 2.2.15 cells to apop- tosis but not HepG2 cells.Although LPS cannot damage normal hepatocytes,it might aggravate hep- atocytes damage when their microenvironment was changed by HBV infection.

Objective To provide experimental evidences for developing a safe and effective re- combinant fowlpox virus which can prevent the infection of HIV-2.Methods A fowlpox virus(FPV) transferring vector was constructed by inserting HIV-2 gag gene to the downstream of a synthetic complex promoter ATI-p7.5?20 of vector pUTA2.Transfection was then carried out,and recombi- nant FPV(rFPV)was screened by 5'-bromo-deoxyuridine(BrdU),genome PCR and western blot detection.Balb/c mice were immunized with rFPV by muscular injection.Anti-HIV-2 antibody, CD4~+ and CD8~+ T-cell count and specific target-killing activity of spleen CTL in immunized mice were analyzed by ELISA,FACS and LDH release assay,respectively.Results A transferring vector pA- gag was constructed and confirmed by amplifying a fragment of 766 bp from the rFPV genome.Mean- while,HIV-2 multi-antibody-specific protein blot(55 000)was detected from the recombinant virus and the HIV-2 specific antibody was detected from the immunized Balb/c mice.HIV-2 specific target- killing activity of spleen CTL was observed in immunized mice.Conclusion A recombinant fowlpox virus expressing HIV-2 structural protein Gag has been obtained,and it can stimulate HIV-2-specific eelluar and humoral immune reactions in mice.

Objective To explore the influence of brain-derived neurotrophic factor (BDNF) on the differentiation of nerve stem cells (NSCs) from neonatal rat hippocampus in vitro and to find new revulsant of NSCs,which can improve the percentage of NSCs differentiating into neurons.Methods Twenty-four hours neonatal rats were selected to obtain hippocampus tissue to culture NSCs in serum-free culture medium by suspending culture.The high pure NSCs were obtained after passing 2 generations.The culture cells were identified as NSCs by staining of nestin,which was NSCs special marker.After passaged three generations,the NSCs were randomly classified into 2 groups:test group and control group.There were 15 pieces per group.There was 2 mL per piece,which contains 1?105 cells.50 g/L fetal bovine serum(FBS) and 20 ?g/L BDNF were added into foundational culture medium in test group;only 50 g/L FBS was added into foundational culture medium in control group.The neurons and their percentage were tested using the immunofluorescence labeling and flow cytometer after 7 days of differentiated cultivation.Results The hippocampus tissue cells grew in globular in serum-free culture medium by suspending culture,which expressed highly positive by nestin immunofluorescence staining.Its purity was above 90%.The percentage of neurone specific enolase(NSE)-positive cells in test group was 60.45%,which was obviously higher than that of control group (23.67%).The difference was significant between 2 groups(?2=27.75 P

Animals ; Insecticides ; toxicity ; Nitric Oxide ; blood ; Organophosphorus Compounds ; Rabbits ; Superoxide Dismutase ; blood ; Vitamin E ; blood

To observe the clinical effect of Yisui decoction plus western medicine in treating multiple system atrophy patients, totally 65 patients from China-Japan Friendship hospital during 2008-2012 with complete clinical data and received consecutive traditional Chinese medicine and western medicine treatment for more than 3 months were observed changes of traditional Chinese medicine symptom score, part 1 of unified multiple system atrophy rating scale, orthostatic hypotension before treatment and after 3 months treatment. After 3 months treatment, total effective rate of traditional Chinese medicine symptom was 70.8%. Compared with before treatment, score of part 1 of unified multiple system atrophy rating scale was obviously reduced after 3 month treatment (P < 0.001). Ex- cept swallow function without significant improvement, the remaining projects of unified multiple system atrophy rating scale were im- proved obviously (P < 0.05), of which the most obvious differences were orthostatic symptoms, falls and intestinal function (P < 0.001). Orthostatic hypotension after 1 month treatment and 3 month treatment was obviously better than before treatment (P < 0.001). There was no significant difference in orthostatic hypotension between 1 month treatment and 3 month treatment. The research results show that Yisui decoction plus western medicine has a certain effect on improving clinical symptoms of multiple system atrophy patients, especially has a significant effect on orthostatic hypotension, and can maintain a stable clinical effect in a certain period of time.
Adult ; Aged ; Humans ; Hypotension, Orthostatic ; drug therapy ; Male ; Medicine, Chinese Traditional ; adverse effects ; methods ; Middle Aged ; Multiple System Atrophy ; drug therapy ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo evaluate the effect of compound bushen recipe (CBR) in improving the survival state of stress and the overall life span in C. elegans by simulating chronic fatigue syndrome (CFS) under various stress states.

METHODSThe tolerance and the average survival time of adult larvae against heat stress (35 degrees C), oxidative stress (250 microg/mL juglone), and in vivo Abeta protein toxicity (Abeta(1-42) transgenic mutant CL4176) under the intervention of the high (500 mg/L), middle (250 mg/L), and low (100 mg/L) dose CBR were observed. The effect of CBR on the average live time (at 25 degrees C), movement distance in 20 seconds, the frequency of pharyngeal pump in 30 seconds, and the reproductive capability were assessed.

RESULTSCompared with the control group, the survival time of heat stressed C. elegans could be significantly increased in each CBR group (P < 0.01). The survival time of heat stressed C. elegans could be elongated, the protein toxicity be attenuated, and the live time prolonged in the high and middle dose CBR groups (P < 0.01, P < 0.05).The movement distance and the frequency of pharyngeal pump could also be increased in the high dose CBR group (P < 0.01). There was no statistical difference in the reproductive capability among all groups (P > 0.05).

CONCLUSIONSCBR could significantly enhance the stress capacity of C. elegans against internal and external environment, and prolong their lifespan. It did not interfere their normal production, and also could improve the quality of life, thus laying a foundation for further mechanism studies and pharmacological researches on CBR in preventing and treating CFS.

Animals ; Caenorhabditis elegans ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Fatigue Syndrome, Chronic ; drug therapy ; Longevity ; Stress, Physiological

Objective Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel microtiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods Diluted 125Ⅰ-insulin was mixed with 5 ul serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4℃). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1 DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, x2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results (1) The optimized testing condition was described as 2×104 CPM of 125Ⅰ-insulin, 5 ul serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9 %(Kappa value=0.402). There were 25 samples with discordant results, which were positive for IAA titer using the corresponding microtiter plate RIA but negative using the novel RIA kit. (3) In TIDM group the positive rate of IAA was 19.7% (16/71), higher than the healthy controls (0.9%, x2=54.36, P<0.001). The subgroup of T1DM children (with 0-9 years) showed the highest IAA positive rate (55.6% ,x2=4.85, P<0.05). In T2DM group the frequency of IAA was 1.5% (8/551), which had no significant difference comparing with that of healthy controls (x2= 0.95, P >0.05). Conclusions Our proposed microtiter plate RIA method for IAA is highly sensitive and specific, likely to be feasible for clinical application. The frequency of IAA is high in children with T1DM.

Objective To research and analyze nursing measures of phlebitis caused by vein medication Amiodarone Hydrochloride. Methods 150 arrhythmias patients in the Traditional Chinese. Medicine Hospital of Wenling were selected as the subjects. The control group was given routine transfusion nursing, the experimental group was given comprehensive nursing intervention on the basis of the control group, saline irrigation before and after transfusion, observed the local reaction of patients, and controlled the concentration and external application of potato chips. Results After the corresponding nursing measures, in the control group, accounting for 46.7%. 35 cases of phlebitis occurred in the experimental group. The incidence of phlebitis in the experimental group was 16.0%, which was significantly lower than that (46.7%) in the control group, which was statistically significant (P<0.05). In the experimental group, the number of patients with I degree phlebitis was 8, and the proportion was 66.67%. In the control group, there were 7 cases of I degree phlebitis, accounting for 20%, which was statistically significant (P<0.05). In the experimental group, there were 4 cases of II degree phlebitis, the proportion was 33.33%, significantly lower than that (54.28%) of the control group (19 cases of II degree phlebitis) with statistical significance (P<0.05). Conclusion The application of comprehensive nursing measures in arrhythmia patients treated with Amiodarone Hydrochloride vein medication, could significantly reduce the occurrence of phlebitis and improve the curative effect with high safety and clinical significance.

OBJECTIVETo explore the effect of moxibustion at Sanyinjiao (SP 6) on normal process of childbirth and safety of mothers and infants.

METHODSOne hundred and seventy-four cases of singleton primiparas, fetal head position were selected as the subjects and divided into three groups by single-blind randomized method: an acupoint group (n = 59), a non-acupoint group (n = 57) and a blank group (n = 58). When the cervix was dilated by 2 to 3 cm, DAJ-23 type multi-function moxibustion apparatus was used at Sanyinjiao (SP 6) in acupoint group, and at non-acupoint spot (the midpoint from the radial arterial pulse throbbing place on the radial side of the transverse crease of the wrist to the radial end of biceps tendon of the elbow) in non-acupoint group, 30 min for each one. The conventional labor care was adopted in blank group. The differences were observed in the first active stage, the second stage and the third stage of the labor as well as the safety of mothers and infants among three groups.

RESULTSThe durations of the first active stage of labor [(123.05 +/- 96.401) min] and the second stage [(37.407 +/- 22.390) min] in acupoint group were shorter than those [(171.40 +/- 94.339) min, (49.517 +/- 25.373) min] in non-acupoint group, as well as those [(178.79 +/- 97.231) min, (53.684 +/- 20.087) min] in blank group separately (all P < 0.05). It was discovered that the progressions of the first active stage and the second stage of labor were accelerated in acupoint group, and there was no statistically significant difference in the third stage of labor among the three groups (all P > 0.05). Bleeding amount after childbirth in acupoint group was less than those in non-acupoint group and blank group (both P < 0.05), and Apgar scores of newborns in acupoint group and non-acupoint group were higher than that in blank group (both P < 0.05).

CONCLUSIONMoxibustion at Sanyinjiao (SP 6) can shorten the process of childbirth, reduce postpartum bleeding and has no harmful impact on mothers and infants.

Acupuncture Points ; Adult ; Female ; Humans ; Labor, Obstetric ; Moxibustion ; Pregnancy ; Young Adult