[Objective] To explore the in-vitro determination method for the recovery rate of sinomenine in microdialysis probe and to investigate the influencing factors. [Methods] The recovery rate of sinomenine was detected by concentration difference method (by gain and by loss) and zero-net flux method. [Results] The recovery rate detected by increment method was as the same as that by decrement method; the recovery rate had no correlation with the sinomenine concentration in the solvent. Sinomenine recovery assessed by concentration difference method had a good intra-day stability and intra-day reproducibility. Sinomenine concentration and recovery in the solvent could be determined accurately by zero-net flux method. [ Conclusion ] Microdialysis sampling can be used for the pharmacokinetic study of sinomenine and decrement method (ie. retrodialysis) can be used for the determination of recovery rate of sinomenine.

Objective To investigate the therapeutic effects of mesenchymal stem cells (MSC) on rat fulminant hepatic failure (FHF). Methods The rat MSC were separated and purified by adherent culture of whole bone marrow cells. The rat FHF models were established by CCl4 intragastric administration.The rats were divided into experimental group (n=20) and model control group (n=20).And the same dose of saline was administered to rats as normal controls (n=8).Dosage of 1.0 × 106 4',6-diamidino-2-phenylindole (DAPI) labeled MSC were transplanted into rats in experimental group and normal control group through caudal veins,and the same dose of saline was given intravenously in model control group.Part of rats in each group were sacrificed after 7 days and 14 days of injection to evaluate the general condition,survival rate,liver function,tumor necrosis factor (TNF)-α level,liver pathology and MSC homing to the liver between experimental group and model control group.Normal distribution data were compared by independent-sample t test and nonnormal distribution data were analyzed by non-parameter test.ResultsAfter 3 days of injection,the generalcondition of experimental group were better than the model control group. After 7 days of injection,there were 15 and 8 survival rats,the survival rates were statistically different between experimental group and model control group (x2 =4.122,P＜0.05).After 7 days and 14 days of injection,the liver function and TNF-α levels were statistically different between experimental group and model control group (both P ＜0.05),and liver pathology improvement in experimental group was more significant than model control group.DAPI labeled cells increased after transplantation in experimental group, whilefewDAPIlabeledcellswere observedinnormalcontrolgroup.ConclusionsMSC can home to liver of FHF rats after MSC allogeneic transplantation through caudal veins,which can improve liver immunity and liver tissue necroinflammation,and facilitate recovery of liver function.Therefore,it is demonstrated that MSC transplantation has obvious therapeutic effect on rat FHF.

The aim of the present study was to establish a minimally invasive H reflex model in mice for the benefit of the research of clinical spinal cord injury and related diseases. Minimally invasive surgery was performed in hind limb of Kunming mouse under light anesthesia. The skin was incised at the point of one-third of the distance from greater trochanter to the base of the cauda. A pair of fine copper conductors were inserted into the shallow muscle using a syringe needle. After the needles were withdrawed, the retained conductors were ligated and fixed with the tissues surrounding the sciatic nerve as the first pair of stimulating electrodes. Another pair of conductors were inserted and fixed in medial malleolus close to the tibial nerve as the second stimulating electrodes. Copper conductor was inserted passing the skin above the proximal end of the metatarsal and fixed as the recording electrode. The reference electrode was placed at the walking pad in the base of the big toe using the same method. Electromyography (EMG) was used to record M and H waves in planta pedis muscles. The stimulus was a square wave with a width of 0.2 ms and frequency of 0.3 Hz. The latency time of the M and H waves which were induced from the two pairs of stimulating electrodes was recorded. Nerve conduction velocity (NCV) was then calculated from the distance between the cathodes of the stimulating electrodes and the latency time difference of M or H waves. The result showed the achievement ratios of H reflex induction were 92.73% and 81.82% in sciatic and tibial nerves, respectively. The latency time of H wave was about 7~10 ms. Motor nerve conduction velocity (MNCV) obtained was (25.84 ± 4.70) m/s (n = 35), while sensory nerve conduction velocity (SNCV) was (31.45 ± 7.30) m/s (n = 35). The method established in the present paper is simple to practice, does slight harm to the animal, and can produce waveforms with little interference. With these advantages, the method can be applied for the study of the latency of H reflex, and it is suitable for the researches which demands good physical condition of experimental animal during H reflex study. This model can also be applied to the detection of SNCV and MNCV.
Animals ; Electromyography ; H-Reflex ; physiology ; Hindlimb ; innervation ; Mice ; Neural Conduction ; physiology ; Tibial Nerve ; physiology

OBJECTIVETo determine the main factors which affect the percutaneous penetration of artesunate and provide efficient data for the artesunate transdermal delivery system.

METHODTransdermal speed constant and accumulative amount of 12 hours were used for the estimations of various reservior vehicles, and the supplement orthodox design was used to study the effect of pH, various proportion of IPA/Water/IPM, and drug concentration.

RESULTDrug concentration and pH were the main factors which affected the percutaneous penetration of artesunate.

CONCLUSIONThe suitable reservior vehicle can prompt the percutaneous penetration of artesunate, and artesunate TTS will be made with further studies.

2-Propanol ; pharmacology ; Administration, Cutaneous ; Animals ; Antimalarials ; administration & dosage ; pharmacokinetics ; Artemisinins ; administration & dosage ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; Hydrogen-Ion Concentration ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Sesquiterpenes ; administration & dosage ; pharmacokinetics ; Skin Absorption ; drug effects

Objective To explore the clinical application of extracorporeal membrane oxygenation (ECMO) assisted cardiopulmonary resuscitation (CPR)in the treatment of critically ill children with cardiac arrest.Methods The clinical data of critically ill children due to cardiac arrest who were treated with ECMO assisted CPR from June 2012 to December 2015 in Affiliated Bayi Children's Hospital,Clinical Medical College in Army General Hospital,Southern Medical University were retrospectively reviewed,and the datas were analyzed by SPSS 20.0 software.Results A total of 17 patients received ECMO assisted CPR treatment,13 cases were male,4 cases were female,aged from 5 hours to 5 years old,the weight ranged from 3 to 16 kg;5 cases survived,and the survival rate 29.41％;12 cases died,the mortality rate 70.59％,of which 5 cases died of hear failure in withdrawal of ECMO,and 7 cases died of complications after withdrawal of ECMO.The age,sex ratio,body weight and other demographic data between 2 groups were not statistically significant (all P ＞ 0.05).There was no significant difference in the primary diseases between 2 groups (P ＞ 0.05).There was no significant difference in CPR time and ECMO support time between 2 groups (t =1.541,0.375,all P ＞ 0.05).Among 11 cases of children with ECMO-related complications,the incidence rate was 64.71％,the incidence of complication in the survival group was significantly less than that in the death group (x2 =8.709,P =0.003).The common complications of ECMO were bleeding,acral necrosis,infection,multiple organ failure,nervous system injury and acute kidney injury.There was no significant difference in the level of lactic acid between the survival group and the death group (P ＞ 0.05) before ECMO support started,but after 24 h of ECMO support,the lactic acid level in the survival group was significantly lower than that in the death group (t =2.896,P =0.014).Conclusions ECMO assisted CPR can improve the survival rate of critically ill children who have cardiac arrest and have no response to the conventional CPR.The serum lactic acid level after 24 h ECMO support has a guiding significance for the prognosis assessment,and ECMO patients' complications are still the most important factor affecting the prognosis of ECMO assisted CPR patients.

Objective The aim of this study was to analyze risk factors which may affect prognosis of patients with hepatits B virus (HBV)-related liver failure,and to construct a model for prognostic evaluation and further assess its predictive ability.Methods In this retrospective cohort study,569 hospitalized patients who were diagnosed with HBV-related liver failure from January 2007 to December 2010 were enrolled.All the patients were followed up and survival analysis was performed using the Kaplan-Meier method.Univariate and multivariate COX proportional hazards regression analyses were applied to variables such as age,sex,complications,biochemical markers,coagulation markers,and HBV DNA levels to construct a model for prognostic evaluation,and 79 independent cases of HBV-related liver failure were used to confirm the model's predictive ability.Accuracy of the constructed model and model for end-stage liver disease (MELD) was evaluated by receiver operating characteristic (ROC) curves.Results The median survival time for all the patients was 59 days.The survival rates at 1,3,6 months were 58.9％,46.2％ and 45.5％,respectively;and survival rates at 1 and 3 years were 44.9％ and 44.5％,respectively.Hepatic encephalopathy,pulmonary infection,upper gastrointestinal bleeding (UGIB),albumin (Alb),aspartate aminotransferase (AST),creatinine (Cr),international normalized ratio (INR) were determined to be independent risk factors (all P＜0.01) which may affect survival of patients with HBV related liver failure.Accordingly,the prognostic index (PI) of the constructed model for prognostic evaluation 4.98 × assignment of hepatic encephalopathy + 4.57 × assignment of pulmonary infection + 4.41 ×assignment of UGIB-9.69 ×lm[Alb (g/L)]+2.46 ×ln[AST (U/L)]+5.18×ln[Cr (mmol/L)]+3.35×ln (INR) 15.36.The area under receiver operating characteristic curve was 0.838 for the constructed model assessing 90-d survival of the patients,and was 0.751 for model for end-stage liver disease,with no significant difference between the two models (Z=1.085,P =0.278).Conclusions Prognosis of patients with HBV-related liver failure can be accurately predicted by the constructed prognostic assessment model,which is consisted of hepatic encephalopathy,pulmonary infection,UGIB,Alb,AST,Cr,and INR as independent risk factors,and is able to predict the 90 d survival.

Objective To investigate the clinical effect of posterior short-segment pedicle screw reduction and fixation without fusion in treatment of thoracolumbar mono-segmental compression fractures.Methods Thirty cases of thoracolumbar mono-segmental compression fractures admitted from January 2009 to February 2010 were assigned to single posterior pedicle screw fixation (screw group,n =15) and posterior pedicle screw fixation with posterolateral fusion (fusion group,n =15) according to random number table.Clinical results in the two groups were assessed based on Cobb' s angle,anterior vertebral body height ratio (％) and Oswestry disability index (ODI) before operation,after operation,prior to the removal of implant and at the latest follow-up.Results All the cases were followed up for average 24 months.Both operation time and blood loss were less in screw group than in fusion group [(76.58 ±12.67) min vs (116.29 ± 17.45) min,P ＜ 0.01 ; (287.54 ± 30.76) ml vs (480.34 ± 100.54) ml,P ＜0.01],whereas there were no statistical differences between the two groups in aspects of Cobb' s angle and anterior vertebral body height ratio before and after operation,prior to the removal of implant as well as at the latest follow-up.Moreover,no statistical difference of ODI was noted between the two groups prior to the removal of implant and at the latest follow-up.Conclusion Posterior pedicle screw fixation without bone grafting achieves similar effects with pedicle screw fixation with bone grafting.

Objective To establish a stable primary culture of rat cerebellar granule neurons in vitro for further study the toxic effects of chronic arsenic exposure on cerebellar cells.Methods Cerebellar cortices were taken from brain of Wistar rat 5-7 day old after born under stereoscopic microscope.Single cell suspension was acquired after digestion and washing with trypsin (0.25％) and DNase Ⅰ solution,respectively.Granule cells were purified from other cells by differential velocity adherence method for two times.Rat cerebellar granule neurons were seeded in culture plate pre-coated with poly-L-lysine.Neurons growth,development and synaptic connections were observed daily.The neurons were identified by neuron specific enolase (NSE) immunofluorescence technique.Results The neurons were affixed to the culture plate in 24 hours,in reticular arrangement observed under contrast microscope.Granule cells gradually turned round from oval and outlines became clearer in 2-3 days.In 4-6 days,there were a wide range of synaptic connections among the neurons and a mature nerve cell network formed.A large quantity of cerebellar granule neurons was seen by NSE identification.Few bigger cells such as purkinjes cells and glial cell outlines were also seen in the same visual field.Conclusions This is a successful primary culture method for acquirement of rat cerebellar granule neurons.The method can provide experimental basis for future studies the toxic effects of chronic arsenic exposure on cerebellar cells.

Objective To investigate the effect of aluminum on hair loss induc ed by fluoride in fluorosis mice.Methods Sixty male C57BL mice were divided into four groups according to body mass:control group,fluoride (F) group (F-100 mg/L),aluminum(Al) group(Al3+ 270 mg/L) and F + Al group(F-100 mg/L + Al3+270 mg/L).Mice were killed 1 month and 3 months after the experiment,respectively.Bone F content was detected by ion-selective electrode method.The level of bone Al was measured through inductively coupled plasma emission spectrum.Dental fluorosis and hair loss of mice were evaluated by visual method.Results One month after the experiment,no dental fluorosis and hair loss was found in all four groups.The content of bone F was the highest in F group [(2401.649 + 86.835) mg/kg],and the lowest in A1 group [(427.006 + 11.878) mg/kg].The levels of bone F in F + Al group and control group were (1210.332 + 19.531)mg/kg and (538.001 + 33.337)mg/kg,respectively.The difference was statistically significant between any two groups (all P ＜ 0.05).Three month after the experiment,all mice of F treatment group had dental fluorosis and hair loss(10/10).Alopecia areas were found in the neck and back regions only.There was no hair loss in control group,Al group and F + Al group.No dental fluorosis was found in both control and Al groups.Only 2 mice were found with dental fluorosis in F + Al group.The levels of bone F in F group,F + Al group,control group and Al group were (4098.645 + 58.842),(1888.165 ± 12.187),(876.258 + 14.462) and (662.385 ± 8.966) mg/kg,respectively.The difference was statistically significant between any two groups (all P ＜ 0.05).Conclusions The hair loss is found in fluorosis mice.Hair loss of mice is closely associated with the level of F exposure.Al can prevent the occurrence of hair loss induced by F in mice through reducing the accumulation of F.

OBJECTIVETo observe the difference in therapeutic effect of Chinese herbal granule decocted separately and mingly.

METHODSOne hundred patients of functional dyspepsia of Spleen deficiency and Liver stagnancy type were treated with Jianweishu Granule (JWSG, a self-formulated recipe by the author). Half of the patients received JWSG decocted separately, half of them received that decocted mingly. The therapeutic effects between the two groups were compared. Experimental observation on the effects of differently decocted remedies in rats was also conducted.

RESULTSThe therapeutic effects obtained in the two groups were similar, with respective cure rate of 72% and 70% and the total effective rate 96% in both group. There was insignificant difference between the two groups (P > 0.05). Experimental study all showed no difference between the differently decocted remedies in inhibiting gastric acid, pepsin activity, gastric function regulation, small intestine movement improvement and pain alleviation in rats.

CONCLUSIONThe therapeutic effects of JWSG decocted separately or mingly are the same. This fact provides a scientific basis for clinical use of granule form of single Chinese herbs.

Adult ; Animals ; Diagnosis, Differential ; Drug Combinations ; Drug Interactions ; Drugs, Chinese Herbal ; administration & dosage ; Dyspepsia ; drug therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Mice ; Middle Aged ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar ; Stomach Ulcer ; drug therapy ; Yang Deficiency ; drug therapy