Objective To study the inducing apoptosis of Hedyotis diffusa extract (HDE) on human hepatocarcinoma cell line Bel7402 and its molecular mechanism. Methods Human hepatocarcinoma cell line Bel7402 was used in vitro cell culture, there were 3 groups:control group, HDE group and 5-Fu group, the Bel7402 cells were dealed with them respectively. The apoptosis morphologic changes of human hepatocarcinoma cells were observed by invert microscope. The apoptosis rate was detected with hematoxylin stain by light microscope. Inhibition of Bel7402 cell proliferation was measured by MTT assay. The expression of oncogene Bcl-xl and anti-oncogene p53 of Bel7402 ceils were observed with RT-PCR assay. Results Compared with the control group, the condensation of nuclear, vacuolar degeneration of mitochondria could be found through light microscope. The apoptosis rate of HDE group was remarkably increased compared with that in control group (P

AIM: To explore the activative effect of water extract from cultured mycelium of Paecilomyces cicadae (P. cicadae) on peritoneal macrophages (PM?) and alveolar macrophages (AM?) of rats.METHODS: The rats were randomly divided into four groups: normal control group(Ⅰ), cyclophosphamide (Cy) group (Ⅱ), P. cicadae group(Ⅲ), Cy+ P.cicadae group(Ⅳ).The rats were bred in the same circumstance and were administered with corresponding drugs by subcutaneous injection for 26 days. PM? and AM? of rats were irrigated by normal saline and collected by centrifuge and incubated in a humidified incubator for 2 h at 37 ℃. The activities of acid phosphatase (ACP) and lactate dehydrogenase (LDH) in the PM? and AM? were determined by biochemical methods, and the capability of PM? and AM? in phagocytosis of neutral red were measured by colorimetric method too. RESULTS: After administering P. cicadae to rats, the activities of ACP and LDH in PM? and AM? of normal rats were elevated significantly,and the reduction of the activities of ACP and LDH in PM? and AM? of rats due to Cy was notably antagonized, and the capability of PM? and AM? phagocytizing the neutral red were strengthened significantly.CONCLUSION: P. cicadae can activate the PM? and AM? of rats.

Objective:To explore the suitable mouse strains,and establish stable human-miceX-GVHD model.Methods:This study selected the Nude Mice and NOD/SCID, and gave them sublethal dose of γ-ray irradiation whole body , and then intraperitoneal transplanted human peripheral blood mononuclear cells ( PBMC) to establish xenogeneic acute graft-versus-host disease model.By detection of human T cells in the mice′s tail venous blood,tissues,organs of the infiltration and other indicators (By flow cy-tometry and immunohistochemistry ) ,we compared the human immune cell infiltration rates in the two mouse model and recorded the survival time.Finally,we determined the appropriate strains of mice to establish X-GVHD.Optimization means were transfer ways and the appropriate amount of human PBMC ,and the best time to observe the changes of T cell phenotype and function.Results:The NOD/SCID mouse was more suitable for inducing human-mouseX-GVHD model,and there were no significant differences between intrap-eritoneal injection and intravenous injection.Transfer human PBMC more than 5 ×107 can establish human-mouseX-GVHD model.Using the optimized experimental conditions to establish the human-mouseX-GVHD model,we found the 7-11 days was the best time to observe the changes of T cell phenotype and function ,and the average survival time was(14.16±1.77)days.Conclusion:Human-miceX-GVHD model can be successfully established by intraperitoneal injection of 5×107 human PBMC into NOD/SCID, and the best time to observe the changes of T cell phenotype and function is between 7-11 days.

Objective To investigate whether ultrasound (US)-targeted microbubble (MB) destruction (UTMD) can influence the biofilm and bacteria in morphology.Methods Twenty-hour biofilms of Staphylococcus epidermidis RP62A were treated with US or UTMD.The acoustic intensity was 0.5-1.5W/cm2,the duty cycle was 50％ and the duration was 10 minutes.After treatment,the absorbance values (A570) of biofilms stained with the crystal violet were measured to assess the biofilm density.The biofilms were observed with macroscopy and light microscopy.The biofilms were examined by confocal laserscanning microscopy (CLSM) and scanning electron microscopy (SEM).Results A thick and compact biofilm was observed in the untreated control group,and there were no obvious micropores in biofilms under macrology and light micrology.Although there were no significant changes under macroscopy in both biofilms treated with US only and UTMD with 0.5 W/cm2 acoustic intensity,interestingly,many micropores could be found under microscopy.The diameters of micropores increased with increasing acoustic intensity,and the micropores in biofilms treated with UTMD were bigger than those treated with US-only in the same condition of acoustic intensity (P ＜0.05).The largest diameters of micropores were up to 1 mm in biofilms treated with UTMD using 1.5 W/cm2 (P ＜0.05).The biofilm density (A570 value) decreased with increasing of the acoustic intensity,and the values in UTMD group of 1.5 W/cm2 were the lowest (P ＜ 0.05).Micropores also could be observed under CLSM.There were no obvious dead bacteria in biofilms treated with US and UTMD compared with untreated control group (P ＞0.05).Under SEM,the shape of bacteria in biofilms treated with US and UTMD became irregular,and many rounded projection could be observed in the surface of the bacteria treated with UTMD.Conclusions US and UTMD can produce micropores in biofilms,which might help to promote antibiotic activity against biofilms

Objective To evaluate the levels of oxidative stress in patients undergoing long-term and repetitive exposure to hyperbaric oxygen (HBO) treatment. Methods 16 healthy volunteers and 58 patients with sub-acute sudden hearing loss (SHL) exposed to HBO were included in the study. Oxidative stress indices (malondialdehyde, MDA, advanced oxidation protein products, AOPP; superoxide dismutase, SOD) were measured in peripheral blood samples collected at the 5th,10th, 20th and 30th HBO treatments sessions (PO2 0.18 MPa, 1 session per day and 5 sessions per week) and under normal ambient pressure respectively. Results After 5th,10th, 20th and 30th sessions of HBOT, no relevant differences in these three indices were detected compared to pre-HBO exposure, between healthy volunteers (P > 0.05). Conclusions The long-term repetitive HBO treatment for 0.18 MPa of PO2 and 30 sessions could not affect in particular the response of the oxidative stress in healthy persons and patients with sub-acute SHL. The influence on three indices of patients with abnormal situation of oxidative stress undergoing lower pressure of HBO (0.18 MPa) is under investigation.

OBJECTIVETo evaluate the role of thrombopoietin (TPO) in the pathology of chronic thrombocytopenic disease.

METHODSWe measured the endogenous plasma concentration of TPO in 40 patients with acquired aplastic anaemia (AA) and in 32 patients with idiopathic thrombocytopenic purpura (ITP) by a sensitive Sandwich enzyme-linked immunosorbent assay (ELISA) and compared the results.

RESULTSPlasma TPO concentrations were significantly higher in AA patients (774 +/- 393 pg/ ml) in comparison with healthy control subjects (55 +/- 34 pg/ml, P < 0.001), and there was a significant negative correlation between their plasma TPO concentrations and platelet counts. In patients with ITP, however, the TPO levels were normal or only slightly elevated (73 +/- 36 pg/ml), and there was no significant difference compared with healthy controls (P > 0.05). There was also no relationship between their plasma TPO levels and platelet counts.

CONCLUSIONSTPO levels may be regulated not only by platelets but also by megakaryocytes in AA and ITP, and measurement of TPO levels is useful for diagnosing thrombocytopenia and understanding the pathophysiology of thrombocytopenia.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; Female ; Humans ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood ; RNA, Messenger ; analysis ; Thrombopoietin ; blood ; genetics ; therapeutic use

Objective:To analyze the temporal and spatial trends of maternal mortality in China, and to predict the future situation of maternal mortality.Methods:Taking the national maternal mortality rate in 1991-2018 and the maternal mortality rate in 2009-2018 in various provinces and cities of China as the research objects, using the statistical description analysis method to analyze the changes of time and space of maternal mortality, and using ARIMA time series model to predict the future situation of maternal mortality in China, rural areas and cities.Results:Regarding to the spatial and temporal distribution of maternal mortality, the maternal mortality rate in China generally showed a certain decline trend. In 1991, the maternal mortality rate was 80.0/100 000, and in 2018, China′s maternal mortality rate was 18.3/100 000, 77.1% lower than that in 1991, with an average annual growth rate of-5.3%; In 2009, Tibet′s maternal mortality rate was the highest, 232.2/100 000, and Jiangsu′s maternal mortality rate was the lowest, 5.2/100 000, with a difference of 44.7times After nine years of development, Tibet is still the province with the highest maternal mortality rate in China, which is 56.5/100 000, while Shanghai has the lowest maternal mortality rate, which is 1.4/100 000, with a difference of 40.4times. In 1991, the rural and urban maternal mortality rates were 46.3/100, 000 and 100.0/100, 000, respectively, and the urban-rural mortality rate was 1∶2.16. By 2022, the urban-rural mortality rate in China was 1∶0.95. Regarding the prediction of maternal mortality for the future, the national maternal mortality rate in 2022 is 10.1/100 000, the urban maternal mortality rate is 16.0/100 000, and the rural maternal mortality rate is 15.0/100 000.Conclusion:The maternal mortality rate in China has been greatly reduced, and the gap between urban and rural areas has decreased from 53.7/100 000 in 1991 to 0.7/100 000 in 2022, showing a downward trend. However, from the model prediction results, there is a slight rebound in the urban maternal mortality rate, while the rural maternal mortality rate remains stable, which suggests that the government and the health administration should pay more attention to the growing trend of urban maternal mortality while taking reasonable measures to reduce the rural maternal mortality rate, so as to avoid the rebound of urban maternal mortality rate.

BACKGROUND:The Yi people have used Sambucus adnata Wall to treat fractures,bruises,arthritis,etc.The author's group found that the active site aqueous extract(SAW-A)and dichloromethane extract(SAW-B)can promote fracture healing by promoting the expression of genes related to osteoblast proliferation,differentiation and maturation,differentiation and maturation.However,the therapeutic effect of methanol extract(SAW-C)at its active site on osteoarthritis is unclear. OBJECTIVE:To investigate the therapeutic effect of Sambucus adnata Wall extract on osteoarthritis,and to identify the effective targets of Sambucus adnata Wall extract in the treatment of osteoarthritis by network pharmacology technology. METHODS:A rat osteoarthritis model was replicated by anterior cruciate ligamentectomy and model rats were then treated with methanol extract of Sambucus adnata Wall by gavage for 21 days.On the 21st day after modeling,the knee joint of rats was detected by X-ray,the width of the knee joint was measured,oxidative stress indexes and inflammatory factors levels in serum and joint lavage fluid were detected,the morphology of the knee joint was observed by hematoxylin-eosin staining,full-thickness cartilage and hyaline cartilage thickness were measured,and the content of articular cartilage proteoglycan was observed by safranin O-fast green staining.The"drug-component-disease-target"network was constructed.Gene ontology enrichment analysis and Kyoto encyclopedia of genes and genomes enrichment analysis of effective targets were performed,and protein-protein interaction network maps were constructed using cytoscape software. RESULTS AND CONCLUSION:Sambucus adnata Wall extract could reduce tumor necrosis factor-α level in joint lavage fluid and serum levels of prostaglandin E2 and malondialdehyde,while increase the level of superoxide dismutase;relieve joint swelling caused by osteoarthritis;improve the histopathological state of articular cartilage,maintain the thickness of full-thickness articular cartilage,hyaline cartilage and the area of bone trabeculae in subchondral bone;and effectively prevent the loss of proteoglycans in articular cartilage and have a chondroprotective effect.Network pharmacological results showed that the methanol extract of Sambucus adnata Wall may have some targets related to inhibition of tumor necrosis factor,AKT1,interleukin-6,MAPK3,and SRC as well as inhibition of over-activation of EGFR signaling pathway and AGE-RAGE signaling pathway.

BACKGROUND:Previous studies showed that extracts of Sambucus adnata Wall.have the ability to promote the proliferation and differentiation of osteoblasts,fracture healing,anti-inflammatory and antioxidant effects,which can effectively alleviate the development of osteoarthritis.Vascular endothelial growth factor,on the other hand,is a biomarker for the evaluation of osteoarthritis severity. OBJECTIVE:To investigate the effect and mechanism of two extracts of Sambucus adnata Wall.(methanol extract SAW-ME and dichloromethane extract SAW-DCE)on angiogenesis in osteoarthritis. METHODS:(1)Rat models of osteoarthritis were established using anterior cruciate ligament transection and given SAW-ME and SAW-DCE.A sham group was set as a control.Immunohistochemistry and immunofluorescence were used to detect the changes of articular vascular endothelial growth factor A in joint tissue and vascular endothelial growth factor and"H"type blood vessels in serum of osteoarthritis rats.(2)Vascular endothelial cells EA.hy926 were used as the research object and intervened with SAW-ME and SAW-DCE.Cell proliferation was then detected by MTT assay.Vascular endothelial growth factor was used to induce EA.hy926 cells,and the model of angiogenesis was replicated.Cell scratch assay and tube formation assay were performed to study the role and mechanism.(3)EA.hy926 cells were used for transcriptome sequencing to analyze the characteristic changes of cell differential genes and related functions after SAW-DCE intervention. RESULTS AND CONCLUSION:(1)SAW-ME and SAW-DCE downregulated the expression of vascular endothelial growth factor A in the rat knee cartilage and reduced the formation of"H"type vessels in osteoarthritis rats.SAW-ME could significantly decrease the level of vascular endothelial growth factor in serum of osteoarthritis rats(P<0.05).SAW-DCE could also decrease the level of vascular endothelial growth factor in serum of osteoarthritis rats,but there was no significant change.(2)Both SAW-ME and SAW-DCE significantly inhibited vascular endothelial cell migration and tube formation,and downregulated the expression of Ang1 and Tie2 proteins.(3)Transcriptome sequencing analysis found that abnormal angiogenesis in osteoarthritis was related to the PI3K/AKT signaling pathway.(4)To conclude,SAW-ME and SAW-DCE can inhibit angiogenesis in the rat model of osteoarthritis,and the mechanism may be related to the Ang1/Tie2 and PI3K/AKT signaling pathways.

Objective To assess the clinical diagnostic performance of liver stiffness measurement (LSM) and aspartate transaminase (AST)-to-platelet (PLT) ratio index (APRI) for liver fibrosis in chronic hepatitis B (CHB) patients with alanine aminotransferase (ALT) less than or equal to five times of the upper limit of normal (≤5×upper limit of normal [ULN]).Methods FibroScan,blood routine and liver function test were conducted at the day or one day before liver biopsy in 383 CHB patients with ALT≤5 × ULN.The Scheuer scoring system was used for liver histologic assessment.APRI was calculated.Based on the results of liver pathology,the areas under receiver operating characteristic curve (AUC) of LSM and APRI for diagnosis of liver fibrosis stage were compared.Results The median LSM were 5.10 kPa for S0 fibrosis stage,5.20 kPa for S1,6.60 kPa for S2,10.10 kPa for S3,and 18.80 kPa for S4.The median APRI values were 0.36,0.38,0.63,0.61 and 1.27,respectively.The AUC of LSM were 0.817 for ≥S2,0.891 for ≥S3 and 0.913 for ≥S4.And the AUC of APRI were 0.717 for ≥S2,0.711 for ≥S3 and 0.746 for ≥S4.The cut-offs of LSM values were 6.8 kPa for ≥S2,8.7 kPa for ≥S3,and 10.9 kPa for ≥S4.Conclusion LSM can accurately assess the degree of liver fibrosis in CHB patients with ALT ≤5 × ULN,which is superior to APRI in clinical utility.