AIM:To establish a quality standard for Xiangge Zhengqi Capsule(Herba Pogostemonis,Flos Caryophylli,Cortex Magnoliae officinalis,Rhizoma Atractylodis macrocephalae,etc.). METHODS: Xiangge Zhengqi Capsule was identified by GC.The effective components in Xiangge Zhengqi Capsule were determined by GC.The chromatographic conditions were: CP-sil 5CB capillary column(25 m?0.25 mm,0.4 ?m),FID as the detector,programmed temperature and high pure helium as carrier gas with the flow rate of 1 mL/min. RESULTS: The relevant chromatographic peak in Flos Caryophylli,Cortex Magnoliae officinalis,Herba Pogostemonis,Rhizoma Atractylodis macrocephalae and Folium Perillae were identified by GC.The contents of eugenol,magnolol and honokiol could be determined by GC. CONCLUSION: The method is simple,feasible and repeatable.It can be used as quality supervisory control of Xiangge Zhengqi Capsule.

Objective: To investigate whether lipopolysaccharide induced parkin expression and mitophagy in macrophages.Methods:The murine peritoneal primary macrophages were aseptically isolated from Kunming mice and cultured in complete medium.The mitochondrial membrane potential of macrophages was detected by flow cytometry,after the cells were stimulated with 200 ng/ml LPS and labeled mitochondria with JC-1.The parkin mRNA level of macrophages was detected by RT-PCR, protein levels of parkin and autophagic related protein LC3 Ⅱ and LC3 Ⅰ were determined by Western blot.The distribution and co-localization of parkin with LC3 and mitochondria in macrophages were respectively observed by laser scanning confocal microscope, before and after the cells were treated with LPS.Results: Flow cytometry results after JC-1 staining showed that mitochondrial membrane potential in macrophages was declined after stimulation with 200 ng/ml LPS, and continuously decreased with prolonged treatment time.The mRNA levels of parkin were increased slightly within 6 h after LPS stimulation,but parkin proteins were increased significantly within 6 h after LPS stimulation.The results of parkin distribution showed that parkin was evenly distributed in the cytoplasm at normal status, but became the obvious punctate distribution after LPS stimulation in macrophages.Western blot results showed LC3 Ⅱ/LC3 Ⅰ levels were increased after LPS stimulation, indicating the appearance of macrophage autophagy.Confocal microscopy showed that there were co-localization of parkin,LC3 and mitochondrial in macrophages after LPS stimulation.Conclusion:Parkin expression is increased significantly and mediated mitochondrial autophagy in macrophages after LPS stimulation, which is involved in the clearance of damaged mitochondria,thereby playing a role in regulating macrophage inflammatory response.

Objective To investigate the effect of apolipoprotein E gene knock-out(ApoE KO) and high-fat diet on morphology and the expression of mortalin in hippocampal CA_3 neurons of mice, and to explore the impact of these factors on memory and Alzheimer's disease.Methods Ten wild-type and 10 ApoE KO mice were fed with common chow as the control group and the KO group respectively while 10 ApoE KO mice were fed with high fat diet.Twelve weeks later, the weight and the lid of these mice were measured.The brain tissues were observed using HE staining, nissl staining, protargol staining,immunohistochemistry staining and image analysis by computer.Results In the ApoE KO group, weight,total cholesterol, triglyceride, low-density lipoprotein cholesterol were higher than those in the control group,and these changes were more significant in ApoE KO high-fat diet group.The nissl was higher in the ApoE KO group (0.301±0.031) and in ApoE KO high-fat diet group (0.261±0.020) than those in the control group (0.341±0.035, F=18.068, P<0.05).The mortalin in the ApoE KO group (0.322±0.060) and in ApoE KO high-fat diet group (0.391±0.041) were higher than the control group (0.256±0.061, F=15.230, P < 0.05).Conclusions ApoE KO and high-fat diet can reduce nissl, and improve the expression of mortalin.This protein may be involved in the pathogenesis of Alzheimer's disease.

Objective To identify clinical predictive factors in intractable childhood epilepsy,to provide some evidence for its early diagnosis.Methods Six hundred and twenty-four children with newly diagnosed epilepsy were retrospectively identified in Xinhua Hospital Affiliated to Shanghai Jiaotong University Medical School(1993-2009).The epileptic children were divided into a drug-responsive epilepsy group (n =584) and an intractable epilepsy group (n =40).Clinical data of patients were retrospectively analyzed,including the gender,age,pathogen,inducement,number of pre-treatment seizures,seizure type,seizure type change,seizure duration,family history,previous history,physical examination,brain CT and initial electroencephalogram(EEG) findings.Single factor analysis and Logistic regression analysis were made in 2 groups.Results Strong univariate associations suggested that some factors could increase the risks of intractable epilepsy:symptomatic or cryptogenic epilepsy,mental retardation,type of infantile spasm,positive neurological examination and large absolute number of pre-treatment seizures,and changes in seizure types in the course of the disease.With multiple Logistic regression,independent predictors of intractability were symptomatic or cryptogenic etiology(OR =3.61),large absolute number of pre-treatment seizures and changes in seizure types in the course of the disease(OR =4.76).Conclusions It's necessary to be wary of intractable epilepsy and to adjust therapy accordingly when seizures were uncontrollable and accompanied by one or more conditions such as symptomatic or cryptogenic epilepsy,large absolute number of pre-treatment seizures,changes in seizure types in the course of the disease.

Objective To explore the effect of human leukocyte antigen B* genotype and age on serum homocysteine (Hcy) levels in children with seizures or epilepsy. Methods Fifteen children with seizures or epilepsy in whom HLA-B*15:02 genotype was detected during October 2015 to June 2016 were included. The plasma Hcy concentration in children with different genotypes was compared. The association of Hcy concentration and age was performed by linear-regression analysis. Results The mean concentration of Hcy was 8.38±4.23 μmol/L in children not carrying HLA-B*15:02 gene, which was obviously higher than that in children carrying HLA-B*15:02 gene 13.03±0.97 μmol/L (P<0.05). The Hcy concentration increased with the age (r2 =0.29, P<0.05). Conclusions Elder children with seizures or epilepsy carrying HLA-B*15:02 gene tend to have higher Hcy concentration and increased potential risk of disease. HLA-B*15:02 gene type and age can predict the changes of Hcy concentration in children with convulsions.

AIM To study the effects of No.1 Weiyan Decoction (Codonopsis Radix,Paeoniae Radix alba,Taraxaci Herba,etc.) on hypoxia and hypoxic tolerance in gastric mucosal epithelial cells in rats with precancerous lesion of gastric cancer (PLGC).METHODS Forty rats were randomly divided into normal,model,Vitacoenzyme Tablets and No.1 Weiyan Decoction groups.Except for the normal group,the rats in the other groups were given with N-methyl-N'-nitro-N-nitrosoguanidine and Little Chengqi Decoction to establish a PLGC rat model.Ten weeks after the administration,the gastric mucosal epithelial microvascular ultrastructure was observed,the expressions of Hif1a mRNA,HIF-1 α protein and VEGF protein in gastric mucosal epithelium were detected.RESULTS Compared with the normal group,the number of microvasculatures in gastric mucosal epithelium was decreased;lumen was severely stenosed or expansive;inner wall was not smooth and had adhesion;vascular endothelial cells were swollen;the expression of Hif1α mRNA showed an increasing trend;the expressions of HIF-1α protein and VEGF protein were anomalous in the model group.Compared with the model group,the gastric mucosal vascular lesion of PLGC rats was improved,and the expressions of Hif1a mRNA,HIF-1α protein and VEGF protein were significantly decreased in the No.1 Weiyan Decoction group.CONCLUSION No.1 Weiyan Decoction can improve hypoxia and hypoxia tolerance,and induce angiogenesis in gastric mucosal epithelium in PLGC rats.

Objective: To investigate the inhibitory effect of macrophage foaming by Yinlan Tianzhi formula (YLTZ) and to explain its effects on lipid-induced inflammation and LXRα-ABCA1 signal pathway. Methods: The model of macrophage foaming was induced by incubating the RAW264.7 cells or BMMs with ox-LDL (50 mg·L-1). The serum containing YLTZ was prepared. The cells were divided into blank group, model group, and drug group. After drug intervention, MTT method was used to detect cell proliferation. The lipid accumulation in cells was observed by oil red O staining, and GPO-PAP method was used to determine the total cholesterol content in cells. Protein and mRNA levels were determined by Western blot and RT- qPCR. Results: Compared with control group, after YLTZ treatment, the lipid level was significantly decreased, and the level of mRNA and protein of LXRα and ABCA1 were significant increased. The expression of inflammatory factor COX2 and iNOS was significantly decreased. Conclusion: YLTZ inhibits macrophage foaming through enhancing LXRα-ABCA1 pathway and suppressing of inflammatory response.

Objective : To investigate the effects of hypothyroidism and drug intervention on the cognitive function of rat offspring through the establishment of a rat model of experimental hypothyroidism in pregnancy. Methods : A total of 20 SD female rats were randomly divided into control group ( CON group) and hypothyroid group ( PTU group) . The hypothyroid model was established by propylthiouracil ( PTU) , the thyroid hormone levels of female mice were detected by electrochemiluminescence immunoassay , and the differences between the two groups were compared. After successful modeling , the male mice were mated in cages , and the hypothyroid group was randomly divided into no intervention group ( group I) , first trimester intervention group ( group II) and second and third trimester intervention group ( group III) . The Morris water maze ( MWM) experiment was used to test the learning and memory ability of the rats. The morphological structure of hippocampal neurons , the expression of nucleoprotein (NeuN) and synapse⁃associated protein ( SYN) in mature neurons were observed by hematoxylin⁃eosin staining (HE) , Nissl staining and immunohistochemistry. Results : ① Compared with the CON group , the female mice in the PTU group had a significant increase in TSH and a significant decrease in FT4 (P < 0. 05) . ② In the positioning navigation test , the evasion latency of the pups in each group was gradually shortened. On the 5th day , the incubation period of group I was significantly longer than that of groups CON , II , III (P < 0. 05) . There was little change between groups II and III and CON groups (P > 0. 05) . ③ The residence time of group II was significantly different from that in group I during the space exploration stage (P < 0. 05) . There was a significant difference between the number of pups crossing the platform in group I and groups CON , II and III (P < 0. 05) . ④ There was no significant difference between HE staining and Nissl staining in hippocampal tissues of rats in each group. However, compared with the CON group , the average absorbance of NeuN and SYN proteins in the hippocampus of mice in groups I , II and III was significantly reduced (P < 0. 05) . Conclusion Hypothyroidism will have adverse effects on the cognitive function and hippocampal neuron development of pregnant rats , and the effects of intervention on the cognitive function of offspring at different stages of pregnancy are different , the earlier the intervention , the smaller the damage to cognitive function.

BackgroundSchizophrenia is a common severe mental disorder with complex pathogenesis. There are few studies on the correlation between kynurenine metabolites in peripheral serum and urine in schizophrenia. ObjectiveTo investigate the concentration of tryptophan-kynurenine metabolites and interleukin-6 （IL-6） in serum and urine in patients with schizophrenia， and their correlation with clinical symptoms， so as to explore potential biological characteristics related to schizophrenia. MethodsA total of 38 patients with schizophrenia who met the criteria of the Diagnostic and Statistical Manual of Mental Disorders， fifth edition （DSM-5）， and were hospitalized or attended outpatient clinic at Hangzhou Seventh People's Hospital from December 2021 to December 2022 were included in the study. Additionally， 26 healthy individuals were concurrently recruited from the community of Hangzhou to serve as a control group. All participants were requested to complete the Positive and Negative Symptom Scale （PANSS）. The levels of tryptophan （TRP）， kynurenine （KYN）， kynurenic acid （KYNA）， quinolinic acid （QUIN）， picolinic acid （PIC）， xanthurenate and 5-hydroxytryptamine （5-HT） in both serum and urine were measured using ultra-high-performance liquid chromatography-triple quadrupole linear ion trap mass spectrometry. Serum and urine IL-6 were measured using enzyme-linked immunosorbent assay. Pearson correlation analysis was conducted to examine the correlation between serum and urinary KYN metabolites， as well as the correlation between metabolite levels and clinical symptoms in the patient group. ResultsPatients with schizophrenia had significantly higher level of IL-6 in serum （U=798.500， P<0.01） and lower level of PIC in urine （U=253.000， P=0.013） compared with the control group. Additionally， level of serum KYN was positively correlated with QUIN/KYNA ratio and QUIN/PIC ratio （r=0.562， 0.438， P<0.05） in patients with schizophrenia. 5-HT/KYN ratio in serum was positively correlated with PANSS total score and negative symptom subscale score （r=0.458， 0.455， P<0.01） in patients with schizophrenia. ConclusionSerum TRP-KYN pathway metabolite levels in patients with schizophrenia were associated with neurotoxic metabolite ratios in urine and the severity of negative symptoms. ［Funded by Zhejiang Medical and Health Science and Technology Program Exploratory （number， 2022KY990）］

Objective : To study the effect and mechanism of high glucose on mesothelial-mesenchymal transition (MMT) of peritoneal mesothelial cells (HMrSV5) , and the protective effect of pharmacological blocking of signal transducer and activator of transcription 3 (STAT3) on rat peritoneal fibrosis (PF) model . Methods : The animals were divided into three groups : the sham group , the model group , and the STAT3 inhibitor group . A miniature per- itoneal dialysis catheter was implanted under the dorsal skin of rat and the rat peritoneal fibrosis model was induced by daily injection of high glucose dialysate . After 10 weeks , HE staining was used to evaluate the histology of the peritoneum , and the level of transforming growth factor-β1 (TGF-β1) in the peritoneum was measured by immuno- histochemistry . HMrSV5 was cultured in high glucose and the optimal stimulation concentration of high glucose was determined by Western blot. High glucose was used to stimulate HMrSV5 after successful transfection with si - STAT3 and Western blot was used to measure the protein level of STAT3 , p-STAT3 , and the key enzymes of glycol- ysis 6-phosphofructo-2-kinase/fructose-2 , 6-biphosphatase 3 (PFKFB3) and lactate dehydrogenase A (LDHA) . Results : HE staining showed that administration of STAT3 inhibitor ( BP-1-102) could inhibit the thickening of subperitoneal tissue and the proliferation of vessels in HG dialysis rats . The expression of TGF-β1 in the rats perito- neum of the model group was significantly higher than that in the sham group , and the level of TGF-β1 was marked- ly lower in the STAT3 inhibitor group compared to the model group (P < 0. 05) . Compared to the control group , high glucose induced the up-regulation of α-smooth muscle actin ( α-SMA) , the down-regulation of E-cadherin and STAT3 activation in HMrSV5 (P < 0. 05) . Mesothelial cells treated with high glucose also exhibited high expres- sion of the key enzymes of glycolysis ( PFKFB3 , LDHA) ( P < 0. 05) , and si-STAT3 can effectively inhibit the overexpression of PFKFB3 and LDHA induced by high glucose ( P < 0. 05) . Conclusion STAT3 is involved in high glucose-induced HMrSV5 hyperglycolysis and MMT , and targeting STAT3 alleviates peritoneal fibrosis and an- giogenesis during peritoneal dialysis treatment in rats .