Objective To study the polysaccharides from dried rhizome of Ligusticum chuanxiong.Methods DEAE-Cellulose and gel-filtration chromatography were used to isolate and purify the polysaccharides,whose structures were characterized by chemical and spectral methods.Results Four homogeneous polysaccharides,named LCP-1,LCP-2,LCP-3,and LCP-4 were obtained from the water extract of L.chuanxiong.Four structures were investigated to be complicated polysaccharied fractions,with the molecular weights of 3.1?10~4,5.2?10~4,9.0?10~4,and 3.6?10~4,respectively,then estimated by GPC.Conclusion The four polysaccharides are obtained from this plant for the first time.

Objective To investigate the effects of Qi-Boosting Toxin-Resolving Formula (QBTRF) on CD4+CD25+ regulatory T cells and Th17 cells of patients with middle to late staged nasopharyngeal carcinoma (NPC). Methods Flow cytometry was performed to detect the ratio of CD4+CD25+ regulatory T cells and Th17 cells in the peripheral blood mononuclear cells (PMBC) among 18 patients with middle to late stage of NPC treated by QBTRF added to conventional therapy (treatment group), Foxp3 mRNA and ROR-γt mRNA in PMBC was determined by RT-PCR technique. Furthermore, serum levels of IL-6 and TGF-β were assayed by ELISA. Meanwhile, 15 patients with NPC treated by conventional therapy were taken as the control group. Results The ratio of CD4+CD25+ regulatory T cells to the total CD4+ T cells and the transcriptional level of Foxp3 mRNA in PMBC were significantly lower in treatment group than that of control group (P<0.05), the ratio of Th17 cells to the total CD4+T cells and the transcriptional level of ROR-γt mRNA in PMBC were significantly higher in treatment group than that of control group (P<0.05). However, the serum level of IL-6 was obviously higher in treatment group than that of control group (P<0.05), and the serum leve of TGF-βwas obviously lower in treatment group than that of control group (P<0.05). Conclusion QBTRF can significantly affect the number ratio and functional activity of CD4+CD25+ regulatory T cells and enforce the differentiation of Th17 cells among patients with middle to late staged NPC, which it may be reversed the immune tolerance of NPC through regulating the level of IL-6 and TGF-β.

Objective To study the inhibitory effect of methylmercury chloride(MMC) on rat C6 glioma cells in vitro.Methods The rat C6 glioma cells were cultivated in vitro and divided into control group and MMC-treated group(0.08-10.00 ?mol?L-1 MMC were divided into 8 groups with concentration gradient).MTT assay was performed to evaluate the proliferation inhibitory effect and cytotoxicity effect of MMC with different concentrations on cultured rat C6 glioma cells,and flow cytometry was used to assess the effects of MMC treatment on cell apoptosis and cell cycle in rat C6 glioma cells.Results 1.25,2.50,5.00 and 10.00 ?mol?L-1 MMC could inhibit the proliferation of cultured rat C6 glioma cells in vitro,the viabilities of MMC treated C6 glioma cells were significantly lower than those in control group(P

Objective To investigate the activity of recombinant Vibrio vulnificus hemolysin (rVvhA) on the apoptosis of J774A.1 cells and the related mechanism. Methods The cytotoxic effect of rVvhA on the growth of J774A.1 cells was identified by MTT, celluar and mitochondrial morphology were observed by transmission electron microscopy, apoptosis or necrosis and mitochondrial membrane potential in J774A.1 cells were measured by flow cytometry, activities of caspase-3 ,-8,-9 were detected by spectrophotometry. Results The viability of J774A.1 cells exposed to rVvhA was inhibited, and it is dependent on dose. Celluar and mitochondrial uhrastructure both occurred to change obviously observed by transmission electron microscopy in J774A.1 treated by 2.0 HU/ml and 3.0 HU/ml rVvhA after 8 hours; and 3.0 HU/ml rVvhA group had a better cytotoxic effect on J774A.1 than that of 3.0 HU/ml rVvhA group. The percentage of apoptosis is (7.80±0.62)%, (12.33±0.12)%, respectively. Besides, the mitochondriai membrane potential also reduced, because the rate of fluorescence which is green increase 1.0% (normal) to 9.8% (2.0 HU/ml rVvhA) and 39.2% (3.0 HU/ml rVvhA). At the same time, the caspase-3, -9 activity increased gradually, but caspase-8 remained unchanging. In J774A.1 cells treated by 3.0 HU/ml rV-vhA + caspase-3 inhibitor(Ac-DEVD-FMK) or caspase-9 inhibitor(Ac-LEHD-FMK), The apoptosis of was reduced to(6.23±3.95)% ,(9.60±3.14)%, and the activity of caspase-3, -9 reduced, too. Conclusion The rVvhA has cytotoxic effect on J774A.1. Mitochondria-mediated apoptosis pathway which is dependent on caspase may be related to apoptosis induced by rVvhA in J774A.1.

Objective To construct a recombinant human adenovirus type 3 ( HAd3 ) vector ex-pressing one major epitope of dengue virus type 1.Methods The gene encoding the envelope protein (304-314 aa) of dengue virus type 1 was inserted into the hypervariable region 1 ( HVR1 ) of HAd3 hexon by using overlap PCR.The recombinant gene was cloned into the shuttle plasmid, then linearized with AsisⅠrestriction enzyme and co-transformed into Escherichia coli BJ5183 strains with the digested backbone plas-mid for homologous recombination.The recombinant plasmid pBRAdΔE3GFP-DENV1 was transfected into AD293 cells to rescue recombinant adenovirus strains (rAdΔE3GFP-DENV1).ELISA and Western blot as-say were performed to evaluate the humoral responses induced in BALB/c mice after the immunization with rAdΔE3GFP-DENV1 strains.Results The recombinant adenovirus strains were successfully rescued. ELISA and Western blot assay showed that the antibodies in serum sample could recognize dengue virus type 1 strains.Conclusion The recombinant adenovirus strains expressing the epitope of dengue virus type 1 were successfully constructed.This study provided evidence for the development of multivalent vaccines against dengue virus.

Objective: To detect varicella-zoster virus ( VZV ) antibody levels among children aged 1 to 12 years in Lu'an City, Anhui Province, so as to provide insights into perfection of the varicella immunization strategy. Methods: Children aged 1 to 12 years were recruited from Lu'an City using the stratified random sampling method from July 2018 to February 2019, and subjects' demographics were collected using questionnaires. The inoculation of varicella vaccines was retrieved through the Anhui Immunization Information Management System or review of preventive immunization certificates, and the serum VZV IgG antibody was detected using enzyme-linked immunosorbent assay ( ELISA ). The seroprevalence and geometric mean concentration of the VZV-IgG antibody were estimated, and the changes of serum the VZV-IgG antibody levels were analyzed at different time intervals following varicella vaccination. Results: Totally 734 children were surveyed, with a mean age of ( 6.94±2.95 ) years, and the subjects included 412 boys ( 56.13% ) and 322 girls ( 43.87% ). There were 514 children ( 70.03% ) with a history of varicella vaccination, including 501 children ( 68.26% ) with one dose of varicella vaccine and 13 children ( 1.77% ) with two doses. There were 297 children ( 40.46% ) positive for VZV-IgG antibody, with seroprevalence of 40.46%, and the GMC of VZV-IgG antibody was 74.97 ( 95%CI: 65.55-85.75 ) mIU/mL. The seroprevalence of the VZV-IgG antibody were 34.55%, 42.91%, and 46.15% among the unvaccinated children and children receiving one dose and two doses of varicella vaccine, with the GMCs of 53.04, 86.31 and 114.46 mIU/mL, respectively. The mean time interval between inoculation of the last dose of varicella vaccine and blood sample collection was ( 5.21±2.79 ) years, and the lowest seroprevalene (31.48%) and GMC of the VZV-IgG antibody (49.96 mIU/mL) were found 4 years after inoculation of varicella vaccine. Conclusions The serum VZV-IgG antibody level is low among children aged 1 to 12 years in Lu'an City, and the seroprevalence of the VZV-IgG antibody is affected by age and doses of varicella vaccine. A 2-dose schedule of varicella vaccine is recommended for children.

Objective: To investigate whether Bruton's tyrosine kinase knockout (Btk^-/-) in macrophages attenuates diabetic kidney disease in the streptozotocin (STZ)-induced mice. Methods: Macrophages-specific Btk^-/- mice and control mice (C57BL/6N) were randomly divided into WT group, diabetic group, Btk^-/- group and Btk^-/- diabetic group. The diabetic models were induced by STZ (50 mg/kg). After 12 weeks, relevant biochemical parameters and the histological changes of kidneys were detected. The expression of macrophages marker CD68 were detected by immunofluorescence, and the immunohistochemistry was employed to detect the expression of WT1 and Nephrin on renal podocytes. In addition, the expression of fibronectin (FN), collagen type IV (IV-Col), transforming growth factor-β1 (TGF-β1), iNOS, phospho (p)-Btk, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), MAPK and NF-κB signaling pathway were detected by Western blotting. RT-PCR was used to detect the mRNA of IL-1β, TNF-α and monocyte chemotactic protein-1 (MCP-1). Results: Compared with diabetic group, the mice in Btk^-/- diabetic group had reduced albuminuria and attenuated kidney histopathology significantly, significantly increased WT1 and Nephrin, significantly decreased expression of CD68, FN, IV-Col and TGF-β1, and these changes were correlated with decreased of renal inflammatory cytokines such as IL-1β, TNF-α, MCP-1 and down-regulating MAPK and NF-κB signaling pathway (all P<0.05). Conclusion Macrophages-specific Btk^-/- may protect the kidney of diabetic mice by reducing the expression of renal inflammatory cytokines in MAPK and NF-κB signaling pathway.

Objective To explore relationship between the change of serum N terminal brain natriuretic peptide (NT-proBNP) , central venous oxygen saturation (SCVO2 ) and venous-arterial partial pressure of carbon dioxide [P(v-a)CO2 ] levels and prognosis in elderly patients with septic shock .Methods 94 cases of elderly patients with septic shock ,treated from March 2015 to July 2016 in Intensive Care Unit (ICU) of our hospital ,were chosen as the observation group A ,another 80 cases of nonsevere sepsis patients were chosen as the observation group B ,and a total of 56 patients healthy volunteers with the physical examination at the same peri-od of time in our hospital as normal control group .The serum NT-proBNP level was detected by electrochemiluminescence immuno-assay ,and PvCO2 ,PaCO2 and SCVO2 levels were detected by blood gas analyzer .records of patients with central venous blood car-bon dioxide .The NT-proBNP and P (v-a) CO2 levels in the serum of the three groups were compared between the groups of pa-tients in the observation group (SCVO2 ) .Logistic regression analysis was used to analyze the prognostic factors in elderly patients with septic shock ,and the receiver operating characteristic curve (ROC) was used to analyze the predictive value of NT-proBNP ,P (v-a)SCVO2 and SCVO2 levels .Results The serum NT-proBNP level of the observation group A was higher than that in the the observation group B and the control group (P<0 .05) .The SCVO2 level of the A group was lower than that in the B group ,and the P(v-a)CO2 level of the A group was higher than that in the B group ,the difference was statistically significant (P<0 .05) .The age , NT-proBNP ,P(v-a)CO2 ,lactic acid level and APACHE II score in the death group were higher than those in the survival group , and the difference was statistically significant (P< 0 .05) .SCVO2 level in the death group was lower than that in the survival group ,and the difference was statistically significant (P<0 .05) .Logistic multivariate regression analysis showed that NT-proBNP , P(v-a)CO2 and SCVO2 levels were the risk factors for the prognosis of elderly patients with septic shock .ROC curve showed AUC and the sensitivity of P(v-a)CO2 were higher than those of NT-proBNP ,SCVO2 .Conclusion The levels of serum NT-proBNP ,SC-VO2 and P(v-a)CO2 are related to the prognosis of elderly patients with septic shock ,and P(v-a)CO2 has a high value in the prog-nosis of the patients with septic shock ,and has a good clinical value .

Objective:To compare the location and efficacy of femoral tunnel near-isometric reconstruction of anterior cruciate ligament (ACL) between the transtibial and assisted medial approaches.Methods:The clinical data of 47 patients were retrospectively analyzed who had been admitted by Department of Orthopaedics, The 904 Hospital of PLA for ACL rupture from January 2018 to December 2019. They were divided into 2 groups according to different surgical approaches. In groups A of 21 cases, there were 15 males and 6 females with an age of (29.5 ± 4.8) years and their ACL was reconstructed through the transtibial approach with adjustable Endobutton plate; in group B of 26 cases, there were 18 males and 8 females with an age of (31.2 ± 9.6) years and their ACL was reconstructed through the assisted medial approach with adjustable Endobutton plate. The 2 groups were compared in terms of location of femoral tunnel, Lysholm score and International Knee Documentation Committee (IKDC) score at the last follow-up, and anterior-posterior and rotational stability of the knee joint.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). The 47 patients were followed up for 18 to 27 months (average, 22.3 months). As for the center of the inner opening of the femoral tunnel located by the four grid table method, the X-axis loci was 25.6% ± 2.5% and 26.7% ± 1.8% respectively in groups A and B, showing no statistically significant difference ( P>0.05) while the Y-axis loci 19.8% ± 2.0% and 30.6% ± 1.5% respectively in groups A and B, showing a statistically significant difference ( P<0.05). At the last follow-up, the lyholm scores were 90.9 ± 3.4 and 92.4 ± 3.9 and the IKDC scores 89.9 ± 3.5 and 90.2 ± 3.8 respectively in groups A and B, showing no significant difference between the 2 groups ( P>0.05). There was no significant difference either in the results of front drawer test, Lachman test or axial displacement test between the 2 groups ( P>0.05). Conclusion:In femoral tunnel near-isometric reconstruction of ACL, the transtibial approach can result in a tunnel location which is closer to the top of the condyle than the assisted medial approach, but both approaches can lead to satisfactory curative efficacy in the short postoperative period.

Objective: To investigate the efficacy and safety of anlotinib hydrochloride capsules for the treatment of advanced soft tissue sarcoma based on the data from Department of Bone and Soft Tissue Tumor, Peking University Cancer Hospital&Institute. Methods: Patients were randomized allocated at 2:1 ratio for the anlotinib treatment and placebo group. The treatment group received 12 mg/day of anlotinib for 14 consecutive days in a 21-day cycle. The primary end-point was progression-free survival (PFS), and the secondary end-points were disease control rate (DCR), overall survival (OS), and adverse event rate. Results: A total of 46 patients were enrolled in this study; 7 of them were excluded from per protocol set (PPS). Among the remaining 39 patients, 28 were included in the anlotinib group and 11 in the placebo group. In the anlotinib group, 4 patients had partial remission and 13 had stable disease (SD), whereas in the placebo group, 3 patients had SD. The difference in DCR between the 2 groups was statistically significant (60.7% vs . 27.3%, P=0.082). The DCR of the advanced soft tissue sarcoma in the anlotinib group was 78.6% (11/14). The median PFS in the anlotinib group was 12.4 (95% confidence interval [CI]: 7.6 to 17.2) months, which was significantly longer than 4 months in the placebo group (95% CI: 1.7 to 6.3 months, P=0.043); however, the difference in OS between the 2 groups was not significant (19.4 vs . 17.6 months, P=0.961). Regarding the safety, 2 patients had severe adverse events (7.14%) possibly related with treatment in the anlotinib group; one of them had pneumothorax. The other adverse events were grade 1 to 2. Conclusions: Soft tissue sarcoma is highly responsive to anlotinib, with prolonged PFS. Anlotinib is well tolerated and can be used as a treatment option for advanced soft tissue sarcoma.