OBJECTIVE: To investigate the anti-proliferation effects of andrographolide Derivative(ADA) and its potential anti-tumor mechanisms in MCF-7 cells. METHODS The anti-proliferation effects of ADA were assessed by MTT assay in MCF-7 cells. Apoptotic morphological changes of MCF-7 cells after ADA treatment were observed by high content screening (HCS) after AO/EB double staining and DAPI, Apoptotic number was investigated by using flow cytometry. The expression of Bcl-2, Bax, caspase 3, NF-κB p65, p-IκBɑ and p-IKKβ were measured using Western blot. RESULTS: MTT assay demonstrated that ADA exerted potent anti-tumor effect in a significant concentration-dependent manner;ADA could promote the apoptosis of MCF-7 cells. Western blot results showed that Bcl-2 families and NF-κB p65 and caspase 3 involved in the process of apoptosis, Bcl-2 expression lowered(P<0.05), p-IκBɑ and p-IKKβ expression decreased(P<0.01), Bax, NF-κB p65 and caspase 3 expression increased(P<0.01). CONCLUSION: ADA could inhibit the proliferation of MCF-7 cells and mechanism may be that it might induce apoptosis by regulated the expression of Bax, Bcl-2 and caspase 3 protein and NF-κB passway.

Objective To investigate the functional changes of dendritic cells (DC) in elderly patients with sepsis. Method Elderly patients (n = 20), ages 75 to 86 years, treated in the department of internal medicine for cadres and the medical intensive care unit (MICU), were selected to participate in the study. Patients with ma-ligoant tumors, hematological diseases, immune diseases, or a history of receiving drugs known to interfere with immune functions were excluded. Using the American College of Chest Physicians/Society of Critical Care Medicine (ACCP/SCCM) definition of sepsis, the patients were categorized into four groups: non-sepsis (group A) (n = 5) ; sepsis (group B) (n = 5) ; severe sepsis (group C) (n = 5) ; and septic shock (group D) (n = 5). The peripheral blood mononuclear cells (PBMCs) of each patient were isolated and cultured with human re-combinant granuloceyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) in vitro for 10 days. The cells were examined under an inverted microscope, scanning electron microscopy, and flow cytometry. The MTT colorimetric assay was used to observe the abilities of the dendritic cells to stimulale an allogeneic T lym-phocyte response in vitro. Paired t -test was used to compare changes in the surface markers among the different groups, Results The PBMCs in the four groups of patients differentiated into cells with typical dendritic configura-tions after in vitro cuhure with combined cytokines. The CD40, CD80, CD86, and HLA-DR expressions on the cell surfaces increased after culture,with (43.2±12.5)%/(27.3±9.3)%, (31.4 ± 10.1)%/(22.5 ± 8.7)%, (39.3±15.7)%/(21.9±7.7)%, and (75.4±25.6)%/(58.7±16.7)%, respectively. The stimulation index (the abilities of the dendritic cells to stimulale the allogeneic T lymphocyte response in vitro) in the four groups of patients after culture were (23.3±7.9) in group A, (18.9±8.3) in group B,(11.4±5.1) in Group C,and (5.5 ± 3.7) in Group D. Conclusions The immune functions of the dendritic cells of elderly patients with sepsis decrease in a linear manner with the severity of their septic state.

Objective To discuss the prevalence of non-EV71,non-CA16 virus strains of hand,foot and mouth disease(HFMD) in Guangdong province between 2008 and 2009,and analyze the genetic evolution of these non-EV71,non-CA16 virus strains.Methods Isolated viruses from stool samples collected from outpatient and in-patient cases of HFMD between 2008 and 2009 by human rhabdomyosarcoma(RD) cell and HEp-2 cell,cultures that exhibited a characteristic enterovirus cytopathic effect were evaluated by RT-PCR.Those strains which identified non-EV71,non-CA16 were analyzed by VP1 sequencing and then were identified by BLAST program.A phylogenetic tree was constructed using the Neighor-Joinning method in the MEGA 4.0 software.Results Twenty-two virus strains of non-EV71,non-CA16 were obtained,and nine of the twenty-two virus strains in 2008 were classified into CA2,CA4,and CB3 by BLAST; thirteen of the twenty-two virus strains in 2009 were classified into EV80,Echo13,Echo30,CBS,Echo24,CA10,CA6,and poliovirus 1 by BLAST.The honology of all strains was low,and all the strains belonged to CA,CB,Echoviruses,Enterovirus and poliovirus subgroup.Conclusion Except for EV71 and CA16 was a major causative agent in prevail of HFMD in Guangdong province between 2008 and 2009,there also existed other subgroup Enterovirus.The other twenty-two strains respectively belonged to CA,CB,Echoviruses,Enterovirus and poliovirus subgroup,and none of those strains was predominant.Muti-species Enterovirus occurred concomitantly.

Objective:To develop a method for the quality control of Cuochuang powder .Methods:Rhei Radix et Rhizoma, An-gelicae Dahuricae Dadix and Saposhnikoviae Radix were identified by TLC .GC was used for the determination of patchouli alcohol , menthol and borneol .Results:The TLC spots were clear without any interference from the negative control .The linear range of pat-chouli alcohol was 0.020 1-0.805 6 mg· ml-1 , that of borneol was 0.010 0-0.401 6 mg· ml-1 , and that of menthol was 0.005 1-0.202 8 mg· ml-1.The average recovery (n=6) was 102.03% (RSD=0.91%), 100.10% (RSD =1.94%) and 103.15%(RSD=1.68%),respectively.Conclusion:The method is simple, accurate and repeatable, which can be used for the quality control of Cuochuang powder .

【Objective】To investigate the therapeutic effect of Qiangxin Prescription(QP)on rabbits with heart failure induced by volume overload.【Methods】Twenty New Zealand male rabbits were randomized into four groups:the control group,QP group(QP 2.6?g/kg),western medicine group(Digoxin 20??g/kg and furosemide 1mg/kg)and QP+western medicine group(QP 2.6?g/kg,Digoxin 20??g/kg and furosemide 1?mg/kg).Rabbit models with heart failure induced by volume overload were established.Six hours after the modeling,the treatment groups were given the corresponding drugs according to the experimental design,the treatment lasting 7 days.Four-channel physiologic instrument was used to examine the heart rate(HR),aortic systolic blood pressure(SBP),aortic diastolic blood pressure(DBP),left ventricular systolic pressure(LVSP),left ventricular diastolic pressure(LVDP),the maximum rate of left ventricular pressure rise(+dp/dt_(max)),the maximum rate of left ventricular pressure decrease(-dp/dt_(max))and the maximum rate of left ventricular pressure change(?dp/dt_(max)).Meanwhile,the serum contents of superoxide dismustase(SOD)and malondialdehyde(MDA)were detected.【Results】The difference was insignificant in the body weight in different groups after the modeling but the body weight in western medicine group was decreased one week after medication(P

Objective: To learn the concentration of target genes of severe acute respiratory syndrome coronavirus 2 ( SARS-CoV-2 ) and the influencing factors in the confirmed cases with coronavirus disease 2019 ( COVID-19 ) in Ningbo. Methods: Demographic information and clinical diagnosis of COVID-19 cases reported in Ningbo from January 21 to February 20, 2020 were collected through China Disease Control and Prevention Information System. The Ct values of ORF1ab and N of the cases were collected through Ningbo Center for Disease Control and Prevention and designated hospitals, and analyzed in terms of gender, age, severity, clinical classification, source of case, sampling time and specimen type. Results: There were 157 confirmed COVID-19 cases, including 56 males ( 35.67% ) and 101 females ( 64.33% ). Sixty-seven cases ( 42.68% ) aged between 40 and 60 years old. The Ct values of ORF1ab and N in females were 29.96±5.28 and 30.38±4.90, which were higher than 27.56±4.94 and 28.03±4.88 in males ( P<0.05 ). The Ct values of ORF1ab and N were the lowest when sampled at 1-7 days after onset ( P<0.05 ), which were 27.84±4.80 and 28.35±4.65. The Ct values of ORF1ab ( rs=0.288, P=0.001 ) and N ( rs=0.296, P=0.001 ) were positively correlated with sampling time. The Ct values of ORF1ab in pharyngeal swab and sputum were 29.19±4.85 and 28.74±6.40, the Ct values of N in pharyngeal swab and sputum were 29.61±4.60 and 29.22±6.10, both without significant differences between different samples ( P>0.05 ). Conclusions Sampling time has a great influence on the Ct values of ORF1ab and N of SARS-CoV-2.

Objective:To investigate the clinical characteristics, treatment effect and prognosis of children with nearly diploid neuroblastoma (NB).Methods:A retrospective analysis of the general clinical characteristics (including age, Gender, risk grouping, location of primary tumor, etc.), laboratory test results, treatment and recent prognosis of NB children with nearly diploidy in bone marrow chromosomes by G-banding technology who admitted to Beijing Children′s Hospital, Capital Medical University from January 2015 to December 2018. Kaplan- Meier method was adopted to calculate survival rate.Univariate analysis was performed using Log- Rank test, and multivariate analysis was conducted with Cox regression model. Results:A total of 43 patients, including 27 males and 16 females, with diagnosis were included, with 14 cases in the hypodiploid group and 29 cases in the hyperdiploid group, and the median age was 35.5 months.The 43 children were all in the high-risk group of International Neuroblastoma Staging System(INSS)-Ⅳ.The primary tumors were mainly located in the retroperitoneal adrenal region (83.7%, 36/43 cases). The largest diameter of the tumors was more than 10 cm (53.5%, 23/43 cases), and often accompanied by 2 or more metastases at the time of consultation.In terms of chromosome karyotype and chromosome karyotype of 14 children in the hypodiploid group was 41-45, the most common karyotype was 45 chromosomes[9 cases(64.3%)]. Among 29 children in the hyperdiploid group of the 47 chromosome karyotypes, 11 cases were common (37.9%). Tumor markers were as follows: neuron enolase (NSE) increased in 41 cases children (95.3%) at first diagnosis, and 25 cases (58.1%)> 370 μg/L; 42 cases (97.7%)had lactate dehydrogenase (LDH). The LDH of children in the hypodiploid group was all> 500 U/L, with 1 case was> 10 000 U/L.Nine cases (20.9%) of MYCN gene were detected by fluorescence in situ hybridization (FISH). Treatment and prognosis: the total course of chemotherapy for 43 patients was 1-12, 19(44.2%) patients received autologous stem cell transplantation, 21 patients (46.5%) received postoperative or autologous radiotherapy or metaiodobenzylguanidine treatment, 28 children developed or relapsed with a median duration of 13.8 months, and 15 cases (34.9%) died.The median follow-up time of the 14 children in the hypodiploid group was 14.9 months (2-38 months), 12 cases progressed or relapsed, and 7 died.The median follow-up of 29 children in the hyperdiploid group was 20.0 months (8.1-51.6 months), with 16 patients progressed or relapsed and 8 cases died. Kaplan- Meier survival analysis illustrated that the 3-year projected event free survival (EFS) rate of 43 children was 18.4%, of which 17.1% were in the hypodiploid group and 29.8% in the hyperdiploid group. Conclusions:Preliminary analysis reveals that children with nearly diploid NB are mostly in the stage Ⅳ high-risk group over the age of 18 months, and 2 or more metastases at the time of consultation.The 3-year estimated EFS of 43 children was 18.4%, and the prognosis was worse in the hypodiploid group.

ObjectiveTo investigate the clinical features and risk factors of patient with Wegener's granulomatosis complicated with pulmonary infection.Methods Patients with Wegener's granulomatosis admitted to our hospital in the past 11 years were retrospectively analyzed.Comparisons between groups were performed by t tests or Fisher test.ResultsPulmonary infection occurred in 27 cases with an incidence rate of 29％.Twenty-six percent of pulmonary infections occurred at the initial diagnosis,and 44％ occurred within 6 months,while 30％ occurred later than 6 months.The clinical manifestations of pulmonary infection were productive cough (89％),hemoptysis (63％),fever and fatigue (56％),chest pain and pactoralgia (33％).The most common causative pathogen were bacteria(59％ ),fungi(37％ ),and tubercle bacillus(37％ ).Sinus infection(P=0.01),hypoproteinemia(P=0.03),hypoimmunoglobulinemia (P=0.007),and methylprednisolone pulse therapy(P=0.002) were the risk factors for pulmonary infection.ConclusionThe occurrence of Wegener's granulomatosis complicated with pulmonary infection is high within 6 months.The most common clinical manifestation is productive cough.The most common causative pathogens are bacteria,tubercle bacillus and fungi.Sinus infection,hypoproteinemia,hypoimmunoglobulinemia,and methylprednisolone pulse therapy are risk factors of pulmonary infection.

Glaucoma is a group of diseases which can threaten and damage the optic nerve and its visual pathway, leading to visual impairment as a result. Glaucomatous optic neuropathy is a chronic disease accompanied by apoptosis of retinal ganglion cells ( RGCs ) , visual field defect and cupping of optic nerve head. The gold standard in functional glaucoma evaluation is standard automated perimetry ( SAP) , but it is often limited to the subjective feelings of the patients. Still, visual electrophysiological techniques cannot replace the conventional inspection, but with its rapid development, it has provided a new strategy for the early diagnosis of glaucoma as a supplement because it can show changes in amplitude and latency before visual field defect. Here we review three special electroretinograms and multifocal focal visually evoked potentials in the early diagnosis of glaucoma.

Objective To study clinical significance of anti-momoner C-reactive protein (anti- mCRP) antibody in systemic lupus erythematosus (SLE) and assess the relationship between serum CRP and anti-mCRP antibody.Methods Enzyme-linked immunosorbent assay (ELISA) was applied to determine serum level of anti-mCRP antibody in 113 pateints with SLE,65 patients with other rheumatic diseases,including primary Sjgren syndrome,rheumatoid arthritis,osteoarthritis,ankylosing spondylitis and systemic sclerosis,and 32 healthy controls.Serum level of CRP was evaluated by turbidimetry.Clinical manifestations and laboratory indicators of the patients were all recorded.Results Serum level of anti- mCRP antibody in SLE patients was significantly higher than that in patients with other rheumatic diseases and healthy controls,respectively (t=2.502 and 5.352,respectively,P 0.05).Titer of anti-mCRP antibody closely correlated with systemic lupus erythematosus disease activity index score (r=0.248,P0.05). Conclusions Level of Anti-mCRP antibody increased significantly in patients with SLE,which associated with disease activity of SLE and can be used as a valuable marker in evaluating activity of SLE.