The serine/threonine protein kinase of Streptococcus suis serotype 2 affects the ability of the pathogen to penetrate the blood-brain barrier.

Streptococcus suis serotype 2 (SS2) is a zoonotic agent that causes meningitis in humans and pigs. However, the mechanism whereby SS2 crosses the microvasculature endothelium of the brain is not understood. In this study, transposon (TnYLB-1) mutagenesis was used to identify virulence factors potentially associated with invasive ability in pathogenic SS2. A poorly invasive mutant was identified and was found to contain a TnYLB-1 insertion in the serine/threonine kinase (stk) gene. Transwell chambers containing hBMECs were used to model the blood-brain barrier (BBB). We observed that the SS2 wild-type ZY05719 strain crossed the BBB model more readily than the mutant strain. Hence, we speculated that STK is associated with the ability of crossing blood-brain barrier in SS2. In vitro, compared with ZY05719, the ability of the stk-deficient strain (Δstk) to adhere to and invade both hBMECs and bEnd.3 cells, as well as to cross the BBB, was significantly attenuated. Immunocytochemistry using antibodies against claudin-5 in bEnd.3 cells showed that infection by ZY05719 disrupted BBB tight junction proteins to a greater extent than in infection by Δstk. The studies revealed that SS2 initially binds at or near intercellular junctions and crosses the BBB via paracellular traversal. Claudin-5 mRNA levels were indistinguishable in ZY05719- and Δstk-infected cells. This result indicated that the decrease of claudin-5 was maybe induced by protein degradation. Cells infected by ZY05719 exhibited higher ubiquitination levels than cells infected by Δstk. This result indicated that ubiquitination was involved in the degradation of claudin-5. Differential proteomic analysis showed that E3 ubiquitin protein ligase HECTD1 decreased by 1.5-fold in Δstk-infected bEnd.3 cells relative to ZY05719-infected cells. Together, the results suggested that STK may affect the expression of E3 ubiquitin ligase HECTD1 and subsequently increase the degradation of claudin-5, thus enabling SS2 to traverse the BBB.

Virulent and Vaccine Strains of Streptococcus equi ssp. zooepidemicus Have Different Influences on Phagocytosis and Cytokine Secretion of Macrophages.

Swine streptococcosis is a significant threat to the Chinese pig industry, and Streptococcus equi ssp. zooepidemicus (SEZ) is one of the major pathogens. SEZ ATCC35246 is a classical virulent strain, while SEZ ST171 is a Chinese attenuated vaccine strain. In this study, we employed stable isotope labeling by amino acids in cell culture and liquid chromatography-mass spectrometry (LC-MS) to determine the differential response of macrophages to infection by these two strains. Eighty-seven upregulated proteins and 135 downregulated proteins were identified. The proteomic results were verified by real-time polymerase chain reaction for 10 chosen genes and Western blotting for three proteins. All differentially abundant proteins were analyzed for their Gene Ontology and Kyoto Encyclopedia of Genes and Genomes annotations. Certain downregulated proteins were associated with immunity functions, and the upregulated proteins were related to cytomembrane and cytoskeleton regulation. The phagocytosis rate and cytokine genes transcription in Raw264.7 cells during SEZ ATCC35246 and ST171 infection were detected to confirm the bioinformatics results. These results showed that different effects on macrophage phagocytosis and cytokine expression might explain the different phenotypes of SEZ ATCC35246 and ST171 infection. This research provided clues to the mechanisms of host immunity responses to SEZ ST171and SEZ ATCC35246, which could identify potential therapy and vaccine development targets.

SILAC and LC-MS/MS identification of Streptococcus equi ssp. zooepidemicus proteins that contribute to mouse brain microvascular endothelial cell infection.

Streptococcus equi ssp. zooepidemicus (SEZ) causes meningitis in both humans and animals. Some dissociative proteins of SEZ are cytotoxic to mouse brain microvascular endothelial cells (mBMECs) and may contribute to the penetration of SEZ across the blood-brain barrier (BBB). In this study, the ability of SEZ to penetrate across an in vitro BBB model was confirmed. We used stable isotope labeling with amino acids in cell culture (SILAC) to label SEZ proteins with heavy or light isotope-tagged amino acids, along with LC-MS/MS to determine which SEZ proteins were involved in interactions with mBMECs. The efficiency of SEZ protein isotope labeling was 94.7 %, which was sufficient for further analysis. Forty-nine labeled peptides were identified as binding to mBMECs, which matched to 25 SEZ proteins. Bioinformatic analysis indicated that most of these proteins were cytoplasmic. These proteins may have functions in breaching the host BBB, and some of them are known virulence factors in other bacteria. Indirect immunofluorescence results indicated that SEZ enolase had binding activity toward mBMECs. Protective test results showed that enolase was a protective antigen against SEZ infection. This research is the first application of SILAC combined with LC-MS/MS to identify SEZ proteins that may contribute to the infection of mBMECs and potentially show functions related to breaching the BBB. The outcomes provide many future avenues for research into the mechanism of SEZ-induced meningitis.

Study of the mechanism of acupuncture and moxibustion in protecting the intestinal mucosal barrier in DSS-induced UC rats based on the IL-9/IL-9R pathway / 针灸推拿医学（英文版）

Objective:To observe the effects of acupuncture and moxibustion on interleukin(IL)-9/IL-9 receptor(IL-9R)in the colon tissue of rats with ulcerative colitis(UC)and investigate the protective mechanism of acupuncture and moxibustion on the intestinal mucosal barrier in UC rats. Methods:Male Sprague-Dawley rats were randomly divided into a normal control(NC)group and a modeling group.UC models were prepared by giving 4%dextran sulfate sodium(DSS)water for 7 d.After the successful construction of the UC rat model,the modeling group was randomly divided into a UC group,a herb-insulated moxibustion(HM)group,and an electroacupuncture(EA)group.HM and EA interventions at bilateral Tianshu(ST25)were performed once a day for 7 d.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes in the colon.The serum concentrations of IL-9,IL-6,IL-1β,and hemoglobin-H(HbH)were determined by enzyme-linked immunosorbent assay.The protein expression levels of IL-9,IL-9R,claudin-2,zonula occludens-1(ZO-1),and occludin in the colon tissue were measured by Western blotting or immuno-histochemistry.Immunofluorescence was used to detect the co-expression of PU.1 and CD4 with the IL-9 protein. Results:Compared with the NC group,the colon tissue of UC rats was severely damaged and ulcerated with congestion and edema,and the colonic histopathological score increased significantly(P<0.01).The serum HbH concentration decreased significantly(P<0.01),while the serum concentrations of IL-9,IL-6,and IL-1β increased(P<0.01).The protein expression of colonic ZO-1 and occludin decreased significantly(P<0.01),while the protein expression of colonic IL-9 and IL-9R increased(P<0.05).The positive co-expression levels of IL-9/PU.1 and IL-9/CD4 increased in the colon tissue(P<0.05).Compared with the UC group,the colonic mucosal structures were gradually repaired in both HM group and EA group,and healed ulcers could be observed,the colonic histopathological score decreased significantly(P<0.05).The serum concentration of HbH increased(P<0.01),while the serum concentrations of IL-9,IL-6,and IL-1β decreased(P<0.05).The protein expression levels of ZO-1 and occludin increased(P<0.05),while the protein expression levels of IL-9 and IL-9R decreased(P<0.01).The positive co-expression levels of IL-9/PU.1 and IL-9/CD4 decreased in the colon tissue(P<0.05). Conclusion:Both HM and EA can inhibit the protein expression levels of IL-9 and IL-9R in the UC colon by regulating the transcription factor PU.1,promote the repair of intestinal mucosal barrier,and down-regulate protein contents of proinflammatory factors IL-9,IL-6,and IL-1β in the serum,which may be one of the key mechanisms of acupuncture and moxibustion in reducing the inflammation of UC colonic mucosa and protecting the intestinal mucosal barrier.

Exploring the mechanism by which acupuncture and moxibustion reduce colonic mucosal inflammation in rats with ulcerative colitis(UC)based on the P2X7R-NLRP3 inflammasome pathway / 针灸推拿医学（英文版）

Objective:To explore the mechanism of acupuncture and moxibustion for ulcerative colitis(UC)from the perspective of the P2X7 receptor(P2X7R)-nucleotide-binding oligomerization domain(NOD)-like receptor protein 3(NLRP3)inflammasome pathway. Methods:Sprague-Dawley rats were randomly assigned to a normal(N)group,a model(M)group,a herb-partitioned moxibustion(HM)group,and an electroacupuncture(EA)group.For modeling,the rats drank 4%dextran sulfate sodium for 7 d.Rats in the HM group and EA group received 7 consecutive days of HM or EA at bilateral Tianshu(ST25),respectively.The histopathological change in colon tissue was observed by hematoxylin-eosin(HE)staining;immunofluorescence staining was used to detect the protein expression of related molecules in the colon tissue,and enzyme-linked immunosorbent assay was used to detect the concentrations or contents of related molecules in the serum and colon tissue.Wild-type(WT)and P2X7R gene knockout(KO)mice were used to construct UC models,histopathological changes in the colon tissue were observed by HE staining,and the NLRP3 protein expression in the colon tissue was observed by immunohistochemistry. Results:Compared with the N group,the colon histopathological score in the M group was significantly increased,and the protein expression of P2X7R,NLRP3,apoptosis-associated speck-like protein containing CARD(ASC),Caspase-1,interleukin-1β(IL-1β),and interleukin-18(IL-18)in the colon tissue and the protein levels of IL-1β and IL-18 in the serum were significantly increased(P<0.05).Compared with the M group,the histopathological scores of the colon in the HM group and the EA group were significantly decreased,and the protein expression levels of P2X7R,NLRP3,ASC,Caspase-1,IL-1β,and IL-18 in the colon tissue and the protein level of IL-18 in the serum were significantly decreased(P<0.05).After UC modeling,the colonic mucosal epithelial damage and inflammatory cell infiltration in P2X7R KO mice were less than those in WT mice,and the NLRP3 protein expression in the colon was also decreased compared with that in WT mice(P<0.05). Conclusion:HM and EA at Tianshu(ST25)may inhibit the protein activities of P2X7R,NLRP3,ASC,and Caspase-1 in the colon tissue of rats with UC,thereby reducing the downstream molecules IL-1β and IL-18 in the P2X7R-NLRP3 inflammasome pathway to relieve UC inflammation.

Therapeutic efficacy of volume-guaranteed high frequency oscillation ventilation on respiratory failure in preterm infants with a gestational age of 28-34 weeks: a prospective randomized controlled study / 中国当代儿科杂志

OBJECTIVES@#To investigate the therapeutic efficacy of volume-guaranteed high frequency oscillation ventilation (HFOV-VG) versus conventional mechanical ventilation (CMV) in the treatment of preterm infants with respiratory failure.@*METHODS@#A prospective study was conducted on 112 preterm infants with respiratory failure (a gestational age of 28-34 weeks) who were admitted to the Department of Neonatology, Jiangyin Hospital Affiliated to Medical School of Southeast University, from October 2018 to December 2022. The infants were randomly divided into an HFOV-VG group (44 infants) and a CMV group (68 infants) using the coin tossing method based on the mode of mechanical ventilation. The therapeutic efficacy was compared between the two groups.@*RESULTS@#After 24 hours of treatment, both the HFOV-VG and CMV groups showed significant improvements in arterial blood pH, partial pressure of oxygen, partial pressure of carbon dioxide, and partial pressure of oxygen/fractional concentration of inspired oxygen ratio (P<0.05), and the HFOV-VG group had better improvements than the CMV group (P<0.05). There were no significant differences between the two groups in the incidence rate of complications, 28-day mortality rate, and length of hospital stay (P>0.05), but the HFOV-VG group had a significantly shorter duration of invasive mechanical ventilation than the CMV group (P<0.05). The follow-up at the corrected age of 6 months showed that there were no significant differences between the two groups in the scores of developmental quotient, gross motor function, fine motor function, adaptive ability, language, and social behavior in the Pediatric Neuropsychological Development Scale (P>0.05).@*CONCLUSIONS@#Compared with CMV mode, HFOV-VG mode improves partial pressure of oxygen and promotes carbon dioxide elimination, thereby enhancing oxygenation and shortening the duration of mechanical ventilation in preterm infants with respiratory failure, while it has no significant impact on short-term neurobehavioral development in these infants.

Investigating the influence of moxibustion on colonic mucosal barrier in rats with dextran sulfate sodium-induced ulcerative colitis / 针灸推拿医学（英文版）

Objective: To observe the effect of moxibustion on the colonic mucosal barrier of rats with ulcerative colitis (UC) induced by dextran sulfate sodium (DSS). Methods: Forty male Sprague-Dawley rats were randomly divided into a normal group and a modeling group, with 20 rats in each group. Rats in the modeling group were subjected to preparing experimental UC models by drinking 4％ DSS for seven consecutive days. Two modeled rats and two normal rats were randomly selected for model identification. After the success of UC model was confirmed, the remaining 18 modeled rats were randomly divided into three groups, a model group, a model + herbal cake-partitioned moxibustion group, and a model + mild moxibustion group, with six rats in each group; the remaining normal rats were randomly divided into three groups, a normal group, a normal + herbal cake-partitioned moxibustion group, and a normal + mild moxibustion group, with six rats in each group. After 7 d of intervention with the herbal cake-partitioned moxibustion or the mild moxibustion, hematoxylin-eosin (HE) staining technique was used to observe the pathological changes of colon tissue under a light microscope; Western blotting and/or immunohistochemical techniques were used to detect the protein expression levels of Occludin, Claudin, junction adhesion molecular 1 (JAM1), mucin 2 (MUC2), and transforming growth factor beta1 (TGF-β1) in rat colon tissue. Results: Compared with the normal group, the colon tissue was severely damaged, the pathological score was significantly increased, and the protein expression levels of Occludin, Claudin, JAM1, MUC2, and TGF-β1 were significantly decreased in the model group (P<0.01); while there were no significant differences in the colonic histopathological score, protein expression levels of Occludin, Claudin, JAM1, MUC2, and TGF-β1 in the normal + herbal cake-partitioned moxibustion group and the normal + mild moxibustion group (P>0.05). Compared with the model group, the model + herbal cake-partitioned moxibustion group and the model + mild moxibustion group showed repaired colon tissue, ulcer healing, significantly reduced pathological score, and significantly increased protein expression levels of JAM1, MUC2, and TGF-β1 (P<0.05); the Occludin protein expression level in the colon tissue of the model + mild moxibustion group was increased (P<0.01). Conclusion: Neither herbal cake-partitioned moxibustion nor mild moxibustion influences the colonic histopathology and intestinal mucosal barrier-related protein expression in the normal rats; both herbal cake-partitioned moxibustion and mild moxibustion can up-regulate the protein expression levels of JAM1, MUC2, and TGF-β1 in the colon tissue of UC rats. Mild moxibustion can up-regulate Occludin protein expression. This may be a mechanism of moxibustion in reducing colonic mucosa inflammation in UC.

Expression and purification of a putative tumor suppressor gene PP5715 in E. coli with growth inhibition of hepatocellular carcinoma cells.

PP5715 is a putative tumor suppressor gene that encodes a protein of 199 amino acids. Expression of PP5715 in E. coli was mainly as inclusion bodies. The recombinant protein was purified by a NTA-Ni2+ affinity column, refolded by dialysis, and shown to suppress growth of two human hepatocellular carcinoma cell lines using the MTT assay or the clonogenic assays. Computer analysis and fusion expression of PP5175 and GFP showed that PP5715 may be a secretory protein. It may bind to receptors on the membrane surface and thus induce regulation of cell growth. These preliminary results suggest that protein PP5715 may be a new tumor suppressor with growth inhibition effects on hepatocellular carcinoma cells.

Effects of moxibustion on the P2X7R/STAT3/VEGF pathway in rats with colitis-associated colon cancer / 针灸推拿医学（英文版）

Objective: To observe the effects of herb-partitioned moxibustion and ginger-partitioned moxibustion on the growth of colon tumors in rats with colitis-associated colon cancer (CACC), and explore the mechanism of moxibustion intervening CACC through the purinergic receptor P2X ligand-gated ion channel 7 (P2X7R)/signal transducer and activator of transcription 3 (STAT3)/vascular endothelial growth factor (VEGF) pathway. Methods: A total of 26 male Sprague-Dawley rats were selected. According to the random number table method, 6 rats were selected as the normal group. The remaining 20 rats were injected intraperitoneally with azoxymethane (AOM) combined with oral dextran sodium sulfate (DSS) to prepare the CACC model. After the model was successfully established, 2 rats were randomly selected for model identification. The remaining 18 rats which were successfully modeled were randomly divided into a model group, a herb-partitioned moxibustion group and a ginger-partitioned moxibustion group, with 6 rats in each group. Moxibustion intervention was performed in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group at Qihai (CV 6) and bilateral Tianshu (ST 25). Moxibustion was performed twice at each point each time, once a day, at a 1-day interval after 6 consecutive interventions, for a total of 30 interventions. After intervention, the colon tumor load, pathological change and histopathological score were observed. Immunohistochemistry was used to detect the expressions of VEGF, P2X7R, phospho-STAT3 (p-STAT3), and nuclear factor-kappa B p65 (NF-κB p65) proteins in rat colon tissue. Western blot was used to detect the levels of p-STAT3 and NF-κB p65 proteins in rat colon tissue. Results: Compared with the normal group, the colon tumor load and histopathological score in the model group were significantly increased (both P<0.001), and different grades of dysplasia were observed in colon tissue from the model group, reaching the degree of adenocarcinoma; the expression level of P2X7R protein in colon tissue was significantly decreased (P<0.001), and the expression levels of p-STAT3, NF-κB p65 and VEGF proteins were significantly increased (all P<0.001) in the model group. Compared with the model group, the colon tumor load, colon histopathological score and the levels of p-STAT3, NF-κB p65 and VEGF proteins in colon tissue were significantly decreased (all P<0.05) in the herb-partitioned moxibustion group and the ginger-partitioned moxibustion group while the expression levels of P2X7R protein in colon tissue were significantly increased (both P<0.05). Conclusion: Both herb-partitioned moxibustion and ginger-partitioned moxibustion can reduce the colon tumor load in CACC rats and delay the progression of colon adenomas. The mechanism may be mediated by the P2X7R/STAT3 pathway to inhibit STAT3 phosphorylation, thereby reducing VEGF protein expression.

The Research of Hyaluronic Acid Functionalized Mesoporous Silica Nanoparticles MCM-41 Loading Paclitaxel for SMMC-7721 Liver Cancer Cells / 中国药学杂志

OBJECTIVE: To establish a drug delivery system based on hyaluronic acid functionalized mesoporous silica nanoparticles MCM-41 loaded with paclitaxel (HA-MCM-41-PTX). The physical and chemical properties, in vitro drug release and the antitumor effect were investigated. METHODS: The morphological structure and particle size of MCM-41 were observed by TEM. The drug delivery system was characterized by PXRD and FTIR. The in vitro release experiments was carried out to investigate the dissolution rate of HA-MCM-41-PTX. The in vitro cells experiment was carried out to explore the mechanism of HA-MCM-41-PTX on cells. RESULTS: The drug loading capacity of HA-MCM-41-PTX was 28.75%. The in vitro drug release experiments showed that HA-MCM-41-PTX exhibited controlled release with a cumulative release of (86.19±5.11)% until 48 h. In vitro cell experiments showed that HA-MCM-41-PTX had excellent targeting effect due to the modification of hyaluronic acid, which was easier to be uptaken by cells and exhibited great antitumor effect. CONCLUSION: HA-MCM-41-PTX is an excellent drug delivery system with both controlled release and targeting antitumor effect.

Application Status and Progress of Mimicking Moxibustion in Clinical Trials of Moxibustion / 世界科学技术-中医药现代化

In recent years, with the widespread clinical application of moxibustion, the clinical trials on moxibustion research are also increasing. However, the establishment of negative control of moxibustion, also called as mimicking moxibustion, has become a major challenge in the randomized controlled trials (RCTs) . In this review, we summarized the establishment methods of mimicking moxibustion in RCTs at home and abroad, and evaluated and elaborated the design and application methods of mimicking moxibustion models from thermal effect, light radiation effect and smoke effect that generated by moxibustion, in order to provide insights for the establishment of an ideal and feasible mimicking moxibustion model in future RCTs of moxibustion.

Protective Mechanism of Preventative Moxibustion at Tianshu Acupoint (ST25) on Intestinal Barrier in UC Rats / 世界科学技术-中医药现代化

Objective: To observe the effect of anti-inflammatory by preventative moxibustion at Tianshu Acupoint (ST25) on UC rats, and to investigate the protective mechanism of preventative moxibustion at Tianshu Acupoint (ST25) onintestinal mucosal barrier in UC rats. Methods: A total of 28 male Sprague-Dawley rats were randomly divided into thenormal group (NC), the ulcerative colitis group (UC), the herb-partitioned moxibustion pretreatment group (HM+P), thewarming moxibustion pretreatment group (WM+P), with 7 in each group. The HM+P and the WM+P were treated withherb-partitioned moxibustion and warming moxibustion for 7 days before the model was established. After pretreatment, the UC, the HM+P and the WM+P were given 4％ dextran sodium sulfate solution for 7 days. Observe the rat pathologicalchanges of colon tissue by HE staining, detect the protein expression in rat colon tissue by immunohistochemistry andwestern blot. Results: compared with the NC group, the pathological scores were increased, the expression of occludin, JAM1, MUC2, ZO-1 were decreased and the expression of NF-κBp65, IL-1βincreased in UC group (P ＜ 0.05);compared with the UC group, the pathological score decreased, the protein expression of occludin, JAM1, MUC2, ZO-1 increased, the protein expression of NF-κBp65, IL-1βdecreased in HM + P group (P ＜ 0.05), the Pathological scoredecreased, the Protein expression of occludin, MUC2, ZO-1 increased, the Protein expression of IL-1β decreased inWM + P group (P ＜ 0.05) . Conclusion: The herb-partitioned moxibustion pretreatment and the warming moxibustionpretreatment could increase the expression of intestinal barrier related proteins in UC rats, it may be one of themechanisms of moxibustion to relieve colonic inflammation of UC.

Herb-partition Moxibustion Relieve Colonic Inflammation in Ulcerative Colitis of DDS-induced Rats Via Dual Inhibition of NF-κB and STAT3 Activation / 世界科学技术-中医药现代化

Objective: To observe the effect of herb-partition moxibustion on colonic inflammation in UC rats, and participate in the regulation of immune regulation via NF-κB pathway and STAT3 phosphorylation. Methods: Rats were randomly divided into normal group, the ulcerative colitis group (UC) group and herb-partition moxibustion group, with 8 rats in each group. The rat model of UC was induced by 4％ DSS. After successful modeling, the rats were treated with moxibustion on bilateral Tianshu acupoints (ST25) . Each acupoint was used with 2 Zhuang moxa, 1 time a day, for 7 times. The effect of the herb-partition moxibustion on UC rats was observed by comparing the histopathological and protein concentrations of serum proinflammatory cytokine. Western Blot was used to detect NF-κB pathway and STAT3 activity in colon tissue. Results: Colonic histopathology in the UC group showed that the mucosal epithelium with ulcer formation and obvious inflammatory response. The herb-partition moxibustion could repair colonic epithelial damage and reduce the inflammatory response of colon tissue in UC rats. Compared with the normal group, the serum IL-1β and IL-6 protein concentrations were significantly increased in the UC group (P ＜ 0.05, P ＜ 0.01), and the STAT3 phosphorylation level and protein expression levels of pIκB-α and NF-κB p65 were significantly increased. The protein expression level of IκB-α was significantly decreased. Compared with the UC group, the serum protein concentrations of IL-1β and IL-6 were significantly lower in the herb-partition moxibustion group (P ＜ 0.05), and protein expression level of NF-κB p65 was decreased in the colon tissue. The phosphorylation level of pIκB-α was decreased, while the protein expression level of IκB-α was increased. Conclusion: Herb-partition moxibustion reduced colonic inflammatory response in UC rats by DSS-induced, the underlying mechanism may related to decrease release of proinflammatory cytokines IL-1β and IL-6 via dual inhibition of NF-κB and STAT3 activation.

Expressions and roles of three inflammatory cytokines in chronic hepatitis B patients with nonalcoholic fatty liver disease / 中华传染病杂志

Objective To investigate the roles of cytokeratin 18 (CK18) M30 and M65,thymosin beta 4 (Tβ4) and tumor necrosis factor (TNF)-α in hepatic steatosis and development of inflammatory and fibrosis in chronic hepatitis B (CHB) patients with nonalcoholic fatty liver disease (NAFLD).Methods A total of 46 CHB patients with NAFLD and 42 CHB patients were collected.Serum CK-18 M30,M65,Tβ4 and TNF-α levels were measured by enzyme linked immunosorbent assay (ELISA) in two groups.The associations between inflammatory factors levels and biochemical or pathological indicators were analyzed.The statistical analysis was conducted by t test and chi square test of two independent samples.The correlation analysis was performed by Pearson and Logistic regression analysis.Results The mean serum CK 18 M30 level in CHB with NAFLD group was (614.48±471.43) U/L,which was significantly higher than that in CHB group (374.50±231.04) U/L (t=2.988,P＜0.01).The mean levels of CK18 M65,Tβ4 and TNF-α in CHB with NAFLD group were (369.41±262.21) U/L,(0.80±0.32) mg/L and (54.87±20.36) ng/L,respectively,and those in CHB group were (296.50±231.44) U/L,(0.68±0.30) mg/L and (51.88± 20.60) ng/L,respectively.There were no difference between CHB with NAFLD group and CHB group (t=1.378,1.810 and 0.685,respectively,all P＞0.05).In CHB with NAFLD patients,the CK-18 M30 level was positively correlated with alanine aminotransferase,triglyceride,fasting blood glucose,histology inflammation score,fibrosis score and steatosis (r=0.507,0.456,0.384,0.551,0.458 and 0.457,respectively,all P＜0.01).Tβ4 level was negatively correlated with inflammation and fibrosis score (r=0.371 and-0.308,respectively,P＜0.05).TNF-α level was positively correlated with inflammation score and steatosis (r=0.570 and 0.441,respectively,P＜0.01).CK-18 M30,Tβ4 and TNF-α were independent predictors of CHB combined with NAFLD,progressive inflammatory fibrosis and severe steatosis.Conclusions Serum CK-18 M30,Tβ4 and TNF-α levels are associated with hepatic steatosis,development of inflammation and fibrosis in CHB with NAFLD patients.

Hepatotoxicity study of extracts and main components in Polygonum multijiorum using high content analysis / 中草药

Objective: A high-content analysis (HCA) method was established for the in vitro evaluation of traditional Chinese medicine and the potential hepatotoxicity components and mechanism of Polygonum multijiorum were discussed. Methods: HepG2 cells were incubated with different concentration and different incubation time of P. multijiorum extracts and components and then cells were stained with three fluorescent probes, then HCA was applied to detect the cell number, nucleus area, mitochondrial mass, and mitochondrial membrane potential. Results: The ethyl acetate extract and dichloromethane extract had served hepatotoxicity. No significant changes were observed at low concentration (0.01 and 1 μmol/L) of P. multijiorum components; However, the cell number of aloe emodin, emodin, rhein, and gallic acid decreased significantly at the concentration of 0.1 μmol/L. Aloe emodin also caused the nuclear swollen. The TC50 values of emodin and rhein obtained from the dose-response curves were similar to the previous reports. Conclusion: The aloe emodin, emodin, rhein, and gallic acid have potential hepatotoxicity that P. multijiorum induces liver injury. The hepatotoxicity of P. multijiorum may be related to mitochondria-mediated apoptosis according to the influence of P.multijiorum components on mitichondrial mass and mitochondrial membrane potential from the results. The HCA is applicable to evaluate the hepatotoxicity of traditional Chinese medicine.

Preformulation study of curcumenol / 国际药学研究杂志

Objective To study the physical- chemical parameters of curcumenol so as to design its optimal preparations. Methods An HPLC method was established to determine the content of curcumenol. Based on the method, the equilibrium solubility, oil/water partition coefficient and stability of curcumenol were investigated. Results The equilibrium solubility in the pure water was 1.93 mg/ml (25 °C) and 2.6 mg/ml (37°), respectively. The oil/water partition coefficient was in the range of 2.2-3.0. Curcumenol solution was unstable at high temperature under light conditions, while pH value showed little effect. Conclusion The analytical method is accurate and reliable. The solubility of curcumenol can be described as strong lipotropic and slightly soluble in water. Nanoformulation and solubilization technologies could improve its bioavailability significantly. Curcumenol should be stored and operated in a dark and cool place.

Thinking and Prospects of Ancient and Modern Researches on Moxibustion Treatment for Inflammatory Bowel Disease (IBD) / 世界科学技术-中医药现代化

By means of searching the ancient and modern literature,this study mainly investigated the etiology and pathogenesis of IBD,summarized the rules of acupoint selection and the experimental mechanism of the moxibustion treatment for IBD,providing the basis and theoretical evidence of acupoint selection and moxibustion therapy for clinical practice and studies.Ancient literatures were retrieved in TCM database system,while modern literatures in Pubmed,CNKI and VIP database from January,2005 to January,2016.Finally,comprehensive analysis was implemented among the selected literatures.It was found that the pathogenesis of IBD was associated with exogenous evils,diet,emotion,and fatigue during chronic diseases in TCM.The treatments of acupuncture and moxibustion for IBD should consider both the primary symptoms and root causes by regulating functional activity of Zang and Fu and also warming and nourishing the spleen and stomach.On this basis,acupoint selection mainly focused on the stomach meridian with the functions of invigorating spleen and stomach,elevating clear qi and descending turbid,recovering the bowel function and strenthening the body and antidiarrhea.Modern research upon the scientific basis of TCM indicated that moxibustion stimulated the thermoreceptors around the acupoints leading to the increased degranulation of mast cells.Immune factors were impacted with the regulation of transcriptional activators in the bowel for repairing mucosal barrier and preventing intestinal fibrosis in IBD.In conclusion,moxibustion was effective on warming and nourishing spleen and stomach and regulating the function activity of bowel for the prevention and treatment of IBD.Achievements were made in the mechanism researches on IBD based on the TCM theory.However,some profound mechanisms need in-depth exploring.

Preformulation study of curcumenol / 国际药学研究杂志

Objective To study the physical-chemical parameters of curcumenol so as to design its optimal preparations. Methods An HPLC method was established to determine the content of curcumenol. Based on the method,the equilibrium solubility, oil/water partition coefficient and stability of curcumenol were investigated. Results The equilibrium solubility in the pure water was 1.93 mg/ml(25℃)and 2.6 mg/ml(37℃),respectively. The oil/water partition coefficient was in the range of 2.2-3.0. Curcumenol solution was unstable at high temperature under light conditions,while pH value showed little effect. Conclusion The analytical method is accurate and reliable. The solubility of curcumenol can be described as strong lipotropic and slightly soluble in water. Nano-formulation and solubilization technologies could improve its bioavailability significantly. Curcumenol should be stored and operated in a dark and cool place.

Effects of the psychological aggression and corporal punishment college students experienced before the age of 10 on anxiety and health related risky behaviors / 中国学校卫生

Objective@#To provide a basis for college students mental health education and guidance, the impact of psychological aggression and corporal punishment college students experienced before the age of 10 on anxiety and health risk behaviors was explored.@*Methods@#In April and May 2019, the convenience sampling method was used to conduct a questionnaire survey among all the freshman and sophomore students of some majors in two universities in Harbin, including a total of 2 150 students. The contents of the questionnaire included general information survey, Parent-Child Conflict Tactics Scale (CTSPC)，Self-Rating Anxiety Scale (SAS) and Adolescent Health related Risky Behavior Inventory (AHRBI).@*Results@#The percentage of college students who suffered psychological aggression and corporal punishment before the age of 10 was 80.05% and 58.37%. The detection rate of corporal punishment in boys and non-only child was significantly higher than that in girls and only child (P<0.05). The students who experienced psychological aggression and corporal punishment before the age of 10 had higher detection rates of anxiety state and six subscales of health risk behaviors than those who didn t experienced those (P<0.05). The Logistic regression analysis showed that, psychological aggression, place of residence, corporal punishment, and only child were positively associated with college students anxiety state (OR=1.85,1.53,1.49,1.34). The Logistic regression analysis showed that, psychological aggression, place of residence were positively associated with college students health-compromising behaviors (OR=1.60,1.26). Psychological aggression, gender factors, corporal punishment, and only child all influenced college students aggression and violence behavior (OR=1.75,1.66,1.63,1.28). Corporal punishment influenced college students suicide and self-injury behavior (OR=3.44). Gender factors and corporal punishment all influenced college students smoking and drinking behavior (OR=2.67,2.25).Corporal punishment and psychological aggression were positively associated with college students rule breaking behavior (OR=1.88,1.55). Corporal punishment, gender factors, and family income all influenced college students unprotected sexual behavior (OR=4.51,1.91,0.62).@*Conclusion@#Parental psychological aggression and corporal punishment college students experienced during childhood have a negative impact on their anxiety and health risk behaviors.

Expression of tumor CD90 in hepatocellular carcinoma and its relationship with epithelial mesenchymal transition / 中华消化杂志

Objective To investigate the expression of stem cell associated protein CD90 in primary hepatocellular carcinoma (HCC) and its relationship with clinical pathological characters ;to analyze the relation between CD90 and epithelial mesenchymal transition (EMT) markers E‐cadherin (E‐cad) and N‐cadherin (N‐cad) .Methods The expressions of CD90 ,E‐cad and N‐cad in 53 tissues of HCC were detected by immunohistochemistry streptavidin‐perosidase (SP ) method , and 10 surrounding of liver benign lesions were studied as control .The expression of E‐cad and N‐cad in CD90 positive cells of two HCC cell lines (M HCC97‐L and HCCLM‐3 ) was measured by flow cytometry .Chi square test and Spearman rank correlation analysis were performed for count data .Independent sample t test was used for measurement data comparison .Results There was no CD90 expression in control liver tissue . The positive rate of CD90 expression in HCC tissue was (34% ,18/53) ,and the difference between two groups was statistically significant (χ2 = 4 .755 , P< 0 .05) .The expression of CD90 in HCC was positively correlated with portal vein cancer embolus (r= 0 .378 , P< 0 .05) and was negatively correlated with histological differentiation degree and capsular infiltration (r= -0 .398 and -0 .519 ,both P<0 .05) .The expression of CD90 was negatively correlated with the expression of E‐cad (r= -0 .341 , P<0 .05) ,however was positively correlated with the expression of N‐cad (r=0 .441 ,P<0 .05) .The positive expression rate of E‐cad in CD90 positive HCCLM‐3 cells was lower than that of MHCC97‐L cells ((2 .56 ± 0 .29)% and (4 .31 ± 0 .18)% ,t= 8 .757 ,P< 0 .05) ,while the positive expression rate of N‐cad was higher than that of MHCC97‐L cells ((8 .10 ± 1 .45)% and (5 .51 ± 0 .44)% ,t= -9 .667 ,P<0 .05) .Conclusions The expression of stem cell associated protein CD90 is correlated with the EMT of HCC .Their interaction promote tumor infiltration metastasis w hich may be a new target of HCC treatment .