Objective To investigate the effects of Saussurea involucrata extract pretreatment on the expression of the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) in focal cerebral ischemia/reperfusion in mice and its possible neuroprotective mechanism.Methods Seventy-two Kunming mice were randomly divided into four groups:sham operation,saline,Saussurea involucrata extract,and edaravone groups (n =18 in each group).Saussurea involucrata extract 0.8 g/kg was given intraperitoneally in the Saussurea involucrata extract group; edaravone 3 mg/kg was given in the edaravone group; and the same volume of saline was given in the saline group.A model of middle cerebral artery occlusion (MCAO) was induced after 7 days of continuous injection.Cerebral infarct volume was determined by 2,3,5-triphenyltetrazolium staining.Immunohistochemical staining was used to detect TLR4-positive cells in ischemic brain tissue.Reverse transcriptase polymerase chain reaction was used to detect the expression of TLR4/NF-κB mRNA.Results The cerebral infarct volume in mice in the saline,Saussurea involucrata extract and edaravone groups was 131.55± 28.25 mm3,84.10 ±13.92 mm3 and 65.10 ± 6.78 mm3,respectively.There were significant difference (F =10.158,P =0.012).The infarct volume in the Saussurea involucrata extract group (P =0.020) and edaravone group (P0.005) was significantly less than that in the saline group,and there was no significantly difference between the 2 groups.The numbers of cortex and TLR4 positive cells in hippocampus area at the ischemic sides in the saline group were significantly more than those in the sham operation group (all P ＜0.001).The numbers of positive cells of cortex and TLR4 in the Saussurea involucrata extract group and the edaravone group were significantly decreased compared to the saline group (all P ＜ 0.05),and there was no significant differences between the Saussurea involucrata extract group and the edaravone group.The expressions of TLR4,p50,and p65 mRNA in the saline group were significantly up-regulated compared to the sham operation group (all P =0.000).Saussurea involucrata extract could significantly down-regulate the expressions of TLR4,p50,and p65 mRNA at 24 hours after ischemia/reperfusion (all P =0.000).Edaravone could significantly down-regulate the expressions of TLR4 and p65 mRNA (all P =0.000) and it had a down-regulated trend for the expression of p50 mRNA (P =0.053); while there was no significant difference in the expressions of TLR4 and p65 mRNA between the Saussurea involucrata extract group and the edaravone group.Conclusions Saussurea involucrata extract pretreatment may significantly reduce the cerebral infarct volume,down-regulate the expressions of TLR4 and NF-κB subunit,and play a neuroprotective effect by inhibiting inflammatory response after ischemia.

Objective To evaluate the effects of the strategy of transmission interruption of schistosomiasis in Runzhou Dis-trict,Zhenjiang City,Jiangsu Province.Methods The comprehensive prevention and control strategy was carried out in Run-zhouDistrict,Zhenjiang City,Jiangsu Province.The strategy was relied mainly on the Oncomelania hupensis snail control,ex-tended chemotherapy of schistosomiasis in residents and the health education.The infection rate of schistosomiasis in residents,area with snails,area with snails controlled,and the rates of awareness and correct behavior of schistosomiasis were as evalua-tion indexes.Results The area with snails controlled was 7 091.50 hm2in Runzhou District,Zhenjiang City from 2001 to 2016.The area with snails reduced year by year from 2001 to 2016.There was a negative correlation between the coverage intensity of snail control and the area with snails(r=-0.874,P=0).There were 1 703 serum positive and 199 fecal positive people of schis-tosomiasis in the permanent residents from 2001 to 2016.These serum and fecal positive people of schistosomiasis were all treat-ed with praziquantel.The serum positive rate of schistosomiasis in the permanent residents dropped to below 1.0％after 2005.The fecal positive patients were not found in 2004 and later.Totally 189 639 people received the questionnaire survey for the knowledge of schistosomiasis control from 2001 to 2016.The rates of awareness and correct behavior of schistosomiasis were raised in the residents year by year.The goal of the transmission interruption of schistosomiasis came to true in Runzhou Dis-trict,Zhenjiang City in 2016.Conclusion The comprehensive prevention and control strategy including mainly the snail con-trol,extended chemotherapy of schistosomiasis and health education could achieve the goal of transmission interruption of schis-tosomiasis in the areas of marshland along the Yangtze River.

Caspases play important roles in cell apoptosis. Measurement of the dynamics of caspase activation in tumor cells not only facilitates understanding of the molecular mechanisms of apoptosis but also contributes to the development, screening, and evaluation of anticancer drugs that target apoptotic pathways. The fluorescence resonance energy transfer (FRET) technique provides a valuable approach for defining the dynamics of apoptosis with high spatio-temporal resolution. However, FRET generally functions in the single-cell level and becomes ineffective when applied in the high throughput detection of caspase activation. In the current study, a FRET sensor was combined with capillary electrophoresis (CE) to achieve a high throughput method for cellular caspase detection. The FRET-based CE system is composed of a homemade CE system and a laser source for detecting the dynamics of caspase-3 in various cells expressing sensors of caspase-3 that have been treated with anticancer drugs, such as cell cycle-independent drug cisplatin and specific cell cycle drugs camptothecin and etoposide, as well as their combination with tumor necrosis factor (TNF). A positive correlation between the caspase-3 activation velocity and drug concentration was observed when the cells were treated with cisplatin, but cells induced by camptothecin and etoposide did not show any apparent correlation with their concentrations. Moreover, different types of cells presented distinct sensitivities under the same drug treatment, and the combination treatment of TNF and anticancer drugs significantly accelerated the caspase-3 activation process. Its high throughput capability and detection sensitivity make the FRET-based CE system a useful tool for investigating the mechanisms of anticancer drugs and anticancer drug screening.
Antineoplastic Agents ; pharmacology ; Camptothecin ; pharmacology ; Caspase 3 ; metabolism ; Cisplatin ; pharmacology ; Drug Screening Assays, Antitumor ; Electrophoresis, Capillary ; methods ; Enzyme Activation ; drug effects ; Etoposide ; pharmacology ; HeLa Cells ; Hep G2 Cells ; Humans ; Neoplasms ; enzymology ; pathology ; Time Factors ; Tumor Necrosis Factor-alpha ; pharmacology

Objective To explore the regulatory mechanism of Macelignan on oxidative stress-mediated neuronal injury in autophagy and apoptosis.Methods Murine hippocampal neuronal HT22 cells were treated with 2.5 mmol·L-1 glutamic acid(Glu)to establish an oxidative stress cell model.The cells were divided into normal group(normal cultured cells),model group(2.5 mmol·L-1 Glu)and experimental-L,-M,-H groups(2.5,5,10 μmol·L-1Macelignan treatment),inhibitor group(2.5 mmol·L-1 Glu+10 μmol·L-1 Macelignan+10 μmol·L-1 LY294002).Aoptosis rate was detected by flow cytometry;the protein expression level of autophagy-related protein LC3B(LC3B),anti-SQSTM1/p62(p62),p21,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)was detected by Western blot.Results The apoptosis rates in the normal group,model group and experimental-L,-M,-H groups were(4.58±1.25)％,(8.75±0.55)％,(6.30±1.71)％,(5.97±2.27)％and(5.49±1.71)％.The difference between model group and normal group was statistically significant(P＜0.01).The difference between experimental-L,-M,-H groups and model group was statistically significant(all P＜0.01).The levels of LC3B in normal group,model group,experimental-L,experimental-M,experimental-H groups and inhibitor group were 0.28±0.02,0.74±0.02,1.02±0.04,0.70±0.03,0.26±0.02 and 0.21±0.01;p62 levels were 0.49±0.08,0.33±0.03,0.50±0.07,0.59±0.01,0.64±0.13 and 0.65±0.06;p21 levels were 0.87±0.02,1.18±0.03,0.98±0.03,0.88±0.03,0.72±0.06 and 0.81±0.02;Bcl-2/Bax levels were 1.74±0.23,1.11±0.10,1.38±0.05,1.66±0.26,1.58±0.29 and 1.53±0.09,respectively.The differences between model group and normal group,between model group and experimental-H group,between model group and inhibitor group,were also statistically significant(all P＜0.01).Conclusion Macelignan can reduce the damage of hippocampal neurons induced by glutamate acid by regulating the process of autophagy and apoptosis,and has obvious neuroprotective effect.

OBJECTIVETo diagnose a Chinese benign familial neonatal convulsions (BFNC) family at the level of gene and investigate its molecular pathogenesis.

METHODSAll family members were studied by clinical examinations and linkage analysis. Mutation analysis of KCNQ2 gene was made by means of polymerase chain reaction (PCR)-direct sequencing and PCR-single strand conformation polymorphism (SSCP) in the proband, 16 family members and 72 unrelated normal individuals.

RESULTSLinkage analysis hinted the linkage of BFNC to KCNQ2, while the linkage to KCNQ3 was excluded. Mutation 1931delG of KCNQ2 gene was found in the proband by DNA-direct sequencing. The same SSCP variant as the proband's was showed in the rest affected members of this family but not in the unaffected members of this family and all of the 72 unrelated normal individuals.

CONCLUSION1931delG of KCNQ2 gene can cause BFNC in China and is novel mutation. The combination of linkage analysis and gene analysis is useful for gene diagnosis.

Epilepsy, Benign Neonatal ; genetics ; Female ; Genetic Linkage ; Humans ; Infant, Newborn ; KCNQ2 Potassium Channel ; KCNQ3 Potassium Channel ; Mutation ; Potassium Channels ; chemistry ; genetics ; Potassium Channels, Voltage-Gated

BACKGROUNDChromosome 13q14 deletion (del13q14), chromosome 1q21 gain (amp1q21) and chromosome 17p13 deletion (del17p13) are the most frequent chromosomal aberrations in multiple myeloma (MM). They play an important role in prognosis. The aim of this study was to investigate the clinical significance of the chromosomal changes in Chinese MM patients.

METHODSInterphase fluorescence in situ hybridization (FISH) on bone marrow (BM) cells was performed in 72 enrolled MM patients. Relationships between chromosomal abnormalities and clinical features, response to therapies and prognosis were analyzed.

RESULTSAs a result of interphase FISH, 77.8% (56/72) patients had chromosome changes. The incidences of each probe were RB1 51.4% (37/72), D13S319 47.2% (34/72), 1q21 45.8% (33/72) and p53 22.2% (12/72). Osteolytic lesion, BM plasma cells index, serum calcium and serum M component were significantly correlated to del13q14. BM plasma cells and hemoglobin were correlated to amp1q21. Serum lactate dehydrogenase (LDH) was correlated with del17p13. Patients with del13q14 treated with bortezomib had a notably higher overall response rate than the patients treated with traditional chemotherapies (93% vs. 65%, P = 0.048). Patients carrying amp1q21 or/and del17p13 did not achieve satisfactory response to bortezomib. The median progression-free survival (PFS) for patients with amp1q21 was 5 months and patients without amp1q21 got 9-month PFS (P = 0.001). The median PFS for patients with del13q14 was 5 months (vs. 8 months, P = 0.026). The median PFS for patients with del17p13 was 3 months (vs. 8 months, P = 0.002). Patients with β(2)-microglobulin > 5.5 mg/L also had a worse outcome, whose median PFS was 5 months (vs. 8 months, P = 0.016).

CONCLUSIONSThe prevalence of chromosomal abnormalities of MM patients was similar in Chinese and Caucasian people. Genetic changes were associated with patients' responses to therapies and prognosis.

Adult ; Aged ; Asian Continental Ancestry Group ; Chromosome Aberrations ; Chromosome Deletion ; Humans ; In Situ Hybridization, Fluorescence ; Interphase ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; genetics ; Prognosis

Objective: To detect the expression of CRLF2 in adult Ph negative acute B lymphocytic leukemia (B-ALL) in newly diagnosed cases, and to investigate the relationship between CRLF2 and the general clinical characteristics, efficacy and prognosis. Methods: 103 cases of newly diagnosed adult B-ALL patients were investigated from Apr 2016 to Dec 2017 in the Department of Hematology, Henan Cancer Hospital. Bone marrow samples was used to detect the expression of CRLF2 in leukemic cells. The expression of CRLF2 ≥20% was defined as CRLF2-high group and <20% was defined as CRLF2-low group. The clinical characteristics and prognosis of the two groups were compared. Results: The Median overall survival (OS) and disease free survial (DFS) in CRLF2-high group were 9.0 months and 4.25 months, respectively. CRLF2-low group were 15.5 months and 10.25 months, respectively. There was a statistically significant difference in median OS and DFS between the two groups (P=0.007, P=0.000) . The 18-month OS and DFS in CRLF2-high group were 38.6% and 25.1%, respectively. CRLF2-low group were 57.8% and 42.3%, respectively. Multivariate analysis showed high expression of CRLF2 was an independent risk factor for OS (HR=2.991, 95% CI 1.429-6.261, P=0.004) and DFS (HR=2.374, 95%CI 1.146-4.960, P=0.041) in patients. Conclusion Patients with high expression of CRLF2 had poor prognosis.

Background Chromosome 13q14 deletion (del13q14), chromosome 1q21 gain (amp1q21) and chromosome 17p13 deletion (del17p13) are the most frequent chromosomal aberrations in multiple myeloma (MM). They play an important role in prognosis. The aim of this study was to investigate the clinical significance of the chromosomal changes in Chinese MM patients.Methods Interphase fluorescence in situ hybridization (FISH) on bone marrow (BM) cells was performed in 72 enrolled MM patients. Relationships between chromosomal abnormalities and clinical features, response to therapies and prognosis were analyzed.Results As a result of interphase FISH, 77.8％ (56/72) patients had chromosome changes. The incidences of each probe were RB1 51.4％ (37/72), D13S319 47.2％ (34/72), 1q21 45.8％ (33/72) and p53 22.2％ (12/72). Osteolytic lesion,BM plasma cells index, serum calcium and serum M component were significantly correlated to del13q14. BM plasma cells and hemoglobin were correlated to amp1q21. Serum lactate dehydrogenase (LDH) was correlated with del17p13.Patients with del13q14 treated with bortezomib had a notably higher overall response rate than the patients treated with traditional chemotherapies (93％ vs. 65％, P=0.048). Patients carrying amp1q21 or/and del17p13 did not achieve satisfactory response to bortezomib. The median progression-free survival (PFS) for patients with amp1q21 was 5 months and patients without amp1q21 got 9-month PFS (P=0.001). The median PFS for patients with del13q14 was 5 months (vs. 8 months, P=0.026). The median PFS for patients with del17p13 was 3 months (vs. 8 months, P=0.002).Patients with β2-microglobulin ＞5.5 mg/L also had a worse outcome, whose median PFS was 5 months (vs. 8 months,P=0.016).Conclusions The prevalence of chromosomal abnormalities of MM patients was similar in Chinese and Caucasian people. Genetic changes were associated with patients' responses to therapies and prognosis.

Objective To analyze the epidemic situation and epidemiological characteristics of malaria in Zhenjiang City from 2004 to 2017, so as to provide the evidence for formulating the reasonable strategies and measures for malaria prevention and control. Methods The epidemiological data of malaria in Zhenjiang City from 2004 to 2017 were collected and analyzed for the epidemic situation, epidemiological characteristics, species of Plasmodium, blood test status for febrile patients. Results A total of 182 malaria cases were reported in Zhenjiang City between 2004 and 2017, 123 cases (67.58%) were infected with P. vivax, and 55 cases (30.22%) were infected with P. falciparum. Among all the reported cases, 41 (22.53%) were local cases, 66 (36.26%) were imported cases from other provinces in China, 75 (41.21%) were imported cases from abroad. Most of the cases were male (133 cases), and 59.89% of them concentrated in the age groups of 20-50 years. These cases were mainly farmers (24.73%, 45/182) and workers (24.18%, 44/182), and most of the cases occurred from July to September. From 2004 to 2017, the blood tests were conducted for 258 879 febrile patients, with a positive detection rate of 0.007% in the local population and 0.724% in floating population, and there was a statistically significant difference between them (P<0.01). Conclusion Since 2011, no indigenous cases have been reported in Zhenjiang City, and therefore, the control of imported malaria from abroad has become the key of malaria prevention and control in this city.

ObjectiveTo analyze the endemic situation of schistosomiasis in national surveillance sites of Zhenjiang City from 2015 to 2019, so as to provide the scientific basis for the consolidation of schistosomiasis control achievements and timely treatment of risk factors. MethodsA total of 7 national schistosomiasis surveillance sites were assigned in Zhenjiang City from 2015 to 2019. According to the National Scheme for Schistosomiasis Surveillance (2014 version), Schistosoma japonicum infections were monitored in local residents, floating populations and livestock, and snail habitats were surveyed in Zhenjiang City from 2015 to 2019. The results of schistosomaisis surveillance in Zhenjiang City from 2015 to 2019 were analyzed. Results There were 15 848 and 7 779 person-times of local residents and floating populations receiving serological screening of schistosomiasis in 7 national surveillance sites of Zhenjiang City from 2015 to 2019, and the seroprevalence rates were 0.95% to 1.75% and 0.14% to 0.91% among local residents and floating populations, respectively; however, no egg-positives were identified. A total of 1 885 herd-times of livestock were detected, and no egg-positives were found. During the 5-year period, snail survey was conducted in snail habitats and suspected snail habitats covering 3 333.79 hm², and there were 108.71 hm² snail habitats identified, including 50.97 hm² re-emerging snail habitats. The density of living snails was 0.007 to 0.125 snails/0.1 m² from 2015 to 2019, which showed a trend for a rise year by year (χ² = 2 732.626, P < 0.01); however, no S. japonicum infections were detected in snails. Conclusions The endemic situation of schistosomiasis has been maintained at a low level in Zhenjiang City, and schistosomiasis control achievements are consolidated. However, snails are widely distributed in the city, and there are still risk factors associated with schistosomiasis transmission. Further integrated interventions require to be reinforced to timely manage the risk of transmission of schistosomiasis and promote the progress towards elimination of schistosomiasis.