OBJECTIVE To study the toxicokinetics and tissue distribution characteristics of alpha-amanitin in rats.METHODS The tail venous blood was collected from SD rats before and 5,10,20,30 and 45 min,1,1.5,2.5,4 and 8 h after intraperitoneal injection of alpha-amanitin(1.5 mg·kg-1),and the concentration of alpha-amanitin in blood was determined by liquid chromatography-mass spectrometry(LC-MS/MS).DAS 2.0 software was used to analyze and plot the drug-time curve with toxicokinetic parame-ters.Based on the toxicokinetics results,18 SD rats were randomly divided into three groups.The rats were sacrificed,and left ventricular arterial(LVA)blood and 9 types of tissue samples involving the heart,liver,spleen,lung,kidney,whole brain,small intestine,stomach wall and testis were collected 15 min,40 min and 2.5 h after dosing,and the concentrations of alpha-amanitin were measured by LC-MS/MS to obtain the tissue distribution results of alpha-amanitin in SD rats.RESULTS Toxicokinetics studies revealed that the peak blood concentration(Cmax)was(633±121)μg·L-1,the elimination half-life(T1/2)was(0.72±0.37)h,and the peak time(Tmax)was(0.52±0.16)h.The total clearance rate(CLz)was(1.62±0.26)L·h·kg-1,the area under the curve(AUC0-t)was(946±183)μg·h·L-1,and the mean reten-tion time(MRT0-t)was(1.18±0.17)h.The apparent volume of distribution(Vz)was(1.65±0.86)L·kg-1.The results of tissue distribution study showed that alpha-amanitin was widely distributed in SD rats with the highest concentration in the kidney,followed by the lung,small intestines,stomach wall,LVA blood and liver,but was low in the heart,spleen,testicles and other tissues,and very low in the brain.Alpha-amanitin was absorbed and eliminated quickly,peaked at 40 min in each tissue,and the concen-tration was minimized after 2.5 h.CONCLUSION The absorption and elimination of alpha-amanitin by intraperitoneal injection are rapid in SD rats,and the blood concentration reaches the peak about 31 min after administration,but can not be detected 4 h later.Alpha-amanitin is mainly distributed in the kidney,followed by the tissues and metabolic organs with rich blood flow,such as the lung,small intestines,stomach wall,LVA blood and liver.The content of alpha-amanitin is low in the heart,spleen,testicles and other tissues,and very low in the brain.It is speculated that it may have toxic targeting effect on the kidney and low blood-brain barrier permeability.

OBJECTIVE: To investigate the protective effects and its possible mechanism of Wuzi Yanzong Pill (WYP) on Parkinson's disease (PD) model mice. METHODS: Thirty-six C57BL/6 male mice were randomly assigned to 3 groups including normal, PD, and PD+WYP groups, 12 mice in each group. One week of intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was used to establish the classical PD model in mice. Meanwhile, mice in the PD+WYP group were administrated with 16 g/kg WYP, twice daily by gavage. After 14 days of administration, gait test, open field test and pole test were measured to evaluate the movement function. Tyrosine hydroxylase (TH) neurons in substantia nigra of midbrain and binding immunoglobulin heavy chain protein (GRP78) in striatum and cortex were observed by immunohistochemistry. The levels of TH, GRP78, p-PERK, p-eIF2α, ATF4, p-IRE1α, XBP1, ATF6, CHOP, ASK1, p-JNK, Caspase-12, -9 and -3 in brain were detected by Western blot. RESULTS: Compared with the PD group, WYP treatment ameliorated gait balance ability in PD mice (P<0.05). Similarly, WYP increased the total distance and average speed (P<0.05 or P<0.01), reduced rest time and pole time (P<0.05). Moreover, WYP significantly increased TH positive cells (P<0.01). Immunofluorescence showed WYP attenuated the levels of GRP78 in striatum and cortex. Meanwhile, WYP treatment significantly decreased the protein expressions of GRP78, p-PERK, p-eIF2α, ATF4, p-IRE1 α, XBP1, CHOP, Caspase-12 and Caspase-9 (P<0.05 or P<0.01). CONCLUSIONS WYP ameliorated motor symptoms and pathological lesion of PD mice, which may be related to the regulation of unfolded protein response-mediated signaling pathway and inhibiting the endoplasmic reticulum stress-mediated neuronal apoptosis pathway.

Objective: To observe the clinical efficacy of acupuncture plus MOTOmed intelligent motor training in treating children with spastic cerebral palsy, and analyze the effects on lower limb motor function, intelligence development level, immune function and cerebral hemodynamics.Methods: A total of 42 children with spastic cerebral palsy were selected as the observation objects, and enrolled into the observation group. Another 42 cases treated in the same period were selected as the control group. Both groups received MOTOmed intelligent motor training, and the observation group was given additional acupuncture therapy, and the control group was given additional conventional rehabilitation treatment. After 2 consecutive treatment courses, the psychomotor development index (PDI) and mental development index (MDI) of Children's Developmental Center of China (CDCC) scale, the scores of gross motor function measure (GMFM) scale and modified Ashworth scale (MAS), and the changes in CD3+, CD4+, CD8+ and CD4+/CD8+ were observed. The peak systolic velocity (PSV) and mean flow velocity (MFV) of the anterior cerebral artery (ACA), middle cerebral artery (MCA) and posterior cerebral artery (PCA) were observed and measured. The clinical efficacy was evaluated. Results: Compared with the same group before treatment, the scores of GMFM, PDI and MDI, levels of CD3+, CD4+ and CD4+/CD8+, PSV and MFV levels of ACA, MCA and PCA in both groups were significantly increased after treatment (all P<0.05), while the CD8+ level had no significant change (both P>0.05). After treatment, the total effective rate of lower limb spasm in the observation group was 90.5％, significantly higher than 71.4％ in the control group (P<0.05). The scores of GMFM, PDI and MDI, the levels of CD3+ and CD4+, PSV and MFV, and the levels of ACA, MCA and PCA in the observation group were all significantly higher than those in the control group (all P<0.05). There were no significant differences in the CD8+ level and CD4+/CD8+ between the groups (all P>0.05). Conclusion: Acupuncture plus MOTOmed intelligent motor training has a better clinical efficacy than conventional rehabilitation plus MOTOmed intelligent motor training in treating children with spastic cerebral palsy, and is also superior in improving lower limb motor function and the level of intellectual development. And the mechanism may be related to the improvement of cerebral hemodynamics.

ObjectiveThere are few reports about abnormal oligonucleotide binding fold domain protein genes (OBGs) affecting the initiation of DNA replication in hepatocellular carcinoma through the microchromosome maintenance (MCM) complex. This study aims to explore the roles of reverse-transcription-related genes (RTGs) in Hepatocellular Carcinoma cells (HCC) and the correlation between gene polymorphisms and abnormal gene expression.Methods We created a mouse model by injecting hepatocellular carcinoma cell line H22 (logarithmic growth phase) and dissected the tumor bodies from tumor-forming mice. The control group was treated by isotonic saline without H22. The healthy liver tissue cells were taken from the control mice. The total RNA of the H22 group and control group were extracted, and differentially expressed genes were analyzed. Screening of differentially expressed reverse transcription-related DEGs (RDEGs), GO and KEGG analysis of RDEGs. The interaction analysis of RDEGs encoded proteins, and the correlation analysis of RDEGs polymorphism and gene expression.ResultsThere were 193 differentially expressed RTGs in HCCs, which were involved in two biological procedures, three cell components, one molecular function, three signal pathways, and three functional sites; Its function is mainly concentrated in DNA replication, especially the construction of MCM complex and telomere complex in which OBGs participate in the initiation of replication. Most related genes had OB fold domains. The results also showed that both AS and SNV caused gene polymorphism was positively correlated with gene expression, and most OBGs in HCC had SNV phenomenon, but not occurred in healthy liver tissue.Conclusion Collectively, AS and SNV may be important regulatory factors for gene expression. SNV may particularly affect the function of OBGs in the MCM complex to abnormally initiate DNA replication in HCC.

OBJECTIVE: To investigate the effect of erythropoietin (EPO) on blood glucoseand plasma insulin level, index of insulin resistance (HOMA-IR), introperitoneal glucose tolerance test (IPGTT), the mRNA and protein level of PR domain-containing 16 (PRDM16), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), fibroblast growth factor 21 (FGF21) of brown adipose tissue (BAT) in mice fed with high fat diet (HFD) in order to provide clues for the mechanism of obesity and complication. METHODS: Twenty C57BL/6J male mice fed with HFD were randomly divided into control group (HFD-Con) and EPO group (HFD-EPO), mice in the two groups were injected intraperitoneally normal saline and EPO (200 IU/kg) res pectively, 3 times per week for consecutive 4 weeks.Then the body weight, blood glucose, plasma insulin level, HOMA-IR and IPGTT were detected.The mRNA and protein level of PRDM16, FGF21, p-STAT3/STAT3 in brown adipose tissue were detected by real-time quantitative RT-PCR and Western blot respectively. RESULTS: After intraperitoneal injection of EPO for 4 weeks, the body weight of the mice in HFD-EPO and HFD-Con groups was (26.65±0.85) g and (31.50±1.6 0) g respectively.The blood glucose of the mice in HFD-EPO group[(62.79±8.09) mg/dl]was significantly decreased compared with that in HFD-Con group[(91.06±9.86) mg/dl].The plasmainsulin level in HFD-EPO group[(10.56±1.06)μU/ml]was significantly decreased compared with that in HFD-Con group[(13.2±1.1)μU/ml, < 0.01].The level of IPGTT in HFD-EPO group was significantly ameliorated and th e HOMA-IR decreased compared with those in HFD-Con group.The mRNA and protein expressions of PRDM16, FGF21 and the level of STAT3 of brown adipose tissue in HFD-E PO group were increased obviously.And there was no difference of FGF21 mRNA content in liver and FGF21 content in plasmabetween the two groups. CONCLUSIONS EPO could promote differentiation of brown adipose tissue by increase in the express ion of PRDM16, and decrease the blood glucose level, ameliorate glucose metabolism in obses mice.
Adipose Tissue, Brown ; Animals ; DNA-Binding Proteins ; Diet, High-Fat ; Erythropoietin ; Fibroblast Growth Factors ; Insulin Resistance ; Male ; Mice ; Mice, Inbred C57BL ; Obesity ; Phosphorylation ; STAT Transcription Factors ; Transcription Factors

Objective:Our study was aimed to analyze the therapeutic effect of early sequential enteral nutrition on postoperative rehabilitation in patients with gastric cancer.Methods:Patients with gastric cancer receiving surgery at our hospital from 2016 to 2017 included and the clinical information was prospective collected and analyzed.Patients were randomly divided into two groups using random number table.Patients in group A were sequentially given amino acid type,short peptide type and then whole protein type,while those in group B received whole protein formulation only.The recovery of gastrointestinal function,postoperative systemic inflammatory response,six-minutes walking test,and enteral nutrition-related complications were compared between the two groups.Results:A total of 71 patients were included in this study (Group A 36 cases,Group B 35 cases).There was no significant difference in terms of the restart anal exhaust between the two groups (P ＞ 0.05).Patients in group A had a significantly shorter postoperative hospitalization (t =4.070;P ＜ 0.01) and the earlier restoration of oral intake than that of Group B (t =3.400;P =0.001).One week after surgery,the levels of CRP (t =2.547;P =0.013) and IL-6 (t =3.172;P =0.002) were significant lower in group A when compared with group B.In addition,patients in group A had a significant higher six minutes walk steps than those in Group B [(416.1 + 36.7) m vs (358.9 ± 32.7) m;t =6.927,P ＜ 0.01].However,no significant difference in enteral nutrition-related complications was found between the two groups (P ＞ 0.05).Conclusion:In patients with gastric cancer,early sequential enteral nutrition can effectively accelerate the postoperative rehabilitation.

OBJECTIVETo investigate the correlation between nonalcoholic fatty liver disease (NAFLD) and three anthropometric indices, namely waist to hip ratio (WHR), body mass index (BMI) and waist to height ratio (WHtR).

METHODSThis retrospective case-control study involved 77 NAFLD patients and 50 patients without such disease, and their data of the 3 anthropometric indices were collected. Risk correlation analysis and Mantel-Haenszel chi-square test were used for correlation analysis.

RESULTSNAFLD was significantly correlated to WHR (chi(2)(MH)=59.609, P<0.001; odds ratio=30.522, 95% CI 12.815-72.695), WHtR (chi(2)(MH)=45.316, P<0.001; odds ratio=21.037, 95% CI 8.665-51.072) and showed a dose-response relationship with BMI (chi(2)=25.017, P<0.001).

CONCLUSIONThese results support a close correlation between NAFLD and the 3 anthropometric indices, indicating that BMI, WHR and WHtR can be significant predictors of NAFLD and have potential value for evaluating and predicting NAFLD.

Adolescent ; Adult ; Anthropometry ; Body Constitution ; Body Height ; Body Mass Index ; Case-Control Studies ; Fatty Liver ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; Retrospective Studies ; Waist Circumference ; Waist-Hip Ratio ; Young Adult

Adult ; Fatty Liver ; diagnosis ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; Waist-Hip Ratio

OBJECTIVETo evaluate the efficacy and safety of Capsule metadoxine in the treatment of alcoholic liver disease.

METHODSA randomized double blind multicenter placebo-controlled clinical study was performed to evaluate the therapeutic effectiveness and safety of capsule metadoxine. Patients in metadoxine group received capsule metadoxine 500mg tid po. Patients in placebo group received placebo 2 pillows tid po. The treatment duration was 6 weeks. Patients were followed up 2 weeks after the treatment. Patients were visited once every 3 weeks during the treatment period. Clinical symptoms and liver function were evaluated in all the patients before treatment, at week 3, week 6 and 2 weeks after therapy. CT scan was done in some patients before treatment and at the end point of therapy.

RESULTS254 patients were recruited in the study, 126 in metadoxine group and 128 in placebo group. Median ALT, AST, GGT level in metadoxine group were decreased from 80.0 U/L, 59.2 U/L, 123.0 U/L (before treatment) to 41.1 U/L, 36.0 U/L, 57.0 U/L (after 6 weeks therapy). The improvement in liver function was more significant in metadoxine group than in placebo group (P less than 0.05). For the patients who stopped drinking during the study, the total effective rate of improvement in liver function was 82.8% in metadoxine group, much higher than that in placebo group (55.7% , P=0.0000). For the patients who did not stop drinking during the study, the total effective rate of improvement in liver function was 65.4% in metadoxine group, which is not significantly higher than that in placebo group (44.8%, P=0.1767). The CT value ratio of liver to spleen was significantly improved in metadoxine group (P=0.0023), and there was no significant difference between the two groups (P=0.6293). The rate of adverse was 1.6% in both of groups.

CONCLUSIONCapsule metadoxine is an effective and safe treatment for alcoholic liver disease.

Administration, Oral ; Adult ; Aged ; Alanine Transaminase ; blood ; Alcohol Deterrents ; administration & dosage ; therapeutic use ; Analysis of Variance ; Aspartate Aminotransferases ; blood ; Capsules ; Double-Blind Method ; Drug Combinations ; Fatty Liver, Alcoholic ; blood ; drug therapy ; pathology ; Female ; Follow-Up Studies ; Humans ; Liver ; diagnostic imaging ; pathology ; Liver Diseases, Alcoholic ; blood ; drug therapy ; pathology ; Liver Function Tests ; Male ; Middle Aged ; Pyridoxine ; administration & dosage ; therapeutic use ; Pyrrolidonecarboxylic Acid ; administration & dosage ; therapeutic use ; Treatment Outcome ; Ultrasonography ; Young Adult ; gamma-Glutamyltransferase ; blood

Cell Line ; Hepatic Stellate Cells ; cytology ; metabolism ; pathology ; Humans ; Oxidative Stress ; Phosphatidylcholines ; pharmacology ; Transforming Growth Factor beta1 ; metabolism