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.

OBJECTIVETo investigate the effect and mechanism of signal transducers and activators of transcription 3 (STAT-3) modulates human tongue squamous cell carcinoma invasion ability via targeting mircoRNA-21.

METHODSTscca and Tca8113P160 human tongue squamous cell carcinoma cell lines were used.WP1066 (STAT-3 inhibitor) , the small molecule inhibitor of STAT-3 was used to suppress the STAT-3 expression. The half maximal inhibitory concentration (IC50 value) of WP1066 in the two cell lines was determined by methyl thiazolyl tetrazolium (MTT) assay. The expression level of STAT-3 and phosphorylation of STAT-3 (pSTAT-3) was examined by Western blotting. Real-time PCR was used to detect the mircoRNA-21 expression after treated with WP1066. Matrigel matrix and transwell assay were used to determine cancer cell colony formation and invasion ability after treated with WP1066. Tumor invasion related proteins in Tscca and Tca8113P160 cell lines were measured by Western blotting. Luciferase reporter gene assay was conducted to detect the relationship between STAT-3 and mircoRNA-21.

RESULTSThe IC50 to WP1066 in Tscca cell was 3.1 and 3.5 µmol/L for Tca8113P160 cell respectively. STAT-3/pSTAT-3 protein level was suppressed significantly (Tscca: STAT-3: F = 887.154, P = 0.000; pSTAT-3: F = 332.212, P = 0.000; Tca8113P160: STAT-3: F = 322.895, P = 0.000; pSTAT-3:F = 788.357, P = 0.000). mircoRNA-21 expression was down-regulated (Tscca:F = 32.157, P = 0.000; Tca8113P160: F = 11.349, P = 0.007). The diameters of culture clone in cell treated with WP1066 were less than control groups (Tscca:F = 15.751, P = 0.004; Tca8113P160: F = 12.964, P = 0.007). The number of tongue cancer cell migrating through the transwell membrane in WP1066 treated group was less than in control groups (Tscca: F = 1688.926, P = 0.000; Tca8113P160: F = 327.528, P = 0.000). In addition, MMP-2/9 protein expression was decreased in both of the cell lines treated with WP1066, while TIMP-3 was up regulated dramatically. STAT-3 could modulate mircoRNA-21 directly.

CONCLUSIONSReduction of STAT-3 can inhibit tongue cancer cell invasion ability via targeting mircoRNA-21.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; MicroRNAs ; genetics ; metabolism ; Phosphorylation ; Pyridines ; pharmacology ; STAT3 Transcription Factor ; antagonists & inhibitors ; metabolism ; Signal Transduction ; Tissue Inhibitor of Metalloproteinase-3 ; metabolism ; Tongue Neoplasms ; genetics ; metabolism ; pathology ; Tyrphostins ; pharmacology

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 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.

Objective To understand the epidemic trend of soil-transmitted nematodiasis in Zhenjiang City from 2006 to 2018, so as to provide evidence for formulating targeted control measures. Methods The permanent residents at ages of 3 years and greater and the immigrants who had lived in Zhenjiang City for more than one year were selected as the study subjects. Fecal samples were collected, and the soil-transmitted nematode eggs were detected in fecal samples with the modified Kato-Katz technique (three smears from one stool sample), while the children at ages of 12 years and lower were tested for Enterobius vermicularis eggs using the adhesive cellophane-tape perianal swab method. The prevalence and intensity of parasitic infections were estimated in Zhenjiang City each year from 2006 to 2018, and the data pertaining to the deworming for soil-transmitted nematode infections were collected during the study period. Results A total of 202 880 person-times were subjected to stool examinations in Zhenjiang City from 2006 to 2018, and 946 persons were detected with soil-transmitted nematode infections, with a mean prevalence of 0.47%. The overall prevalence of human soil-transmitted nematode infections appeared a decline tendency (P < 0.01), with a reduction from 1.10% in 2006 to 0.11% in 2018, and there was a region-specific overall prevalence (P < 0.01), with the highest prevalence seen in Runzhou District (1.65%) and the lowest detected in New District (0.04%). The mean prevalence of human Ascaris lumbricoides, hookworm and Trichuris trichiura infections was 0.33%, 0.09% and 0.04% in Zhenjiang City, respectively, and showed a decline tendency year by year from 2006 to 2018 (P < 0.01). Among all infections, mild infections were predominant. During the period from 2006 through 2018, a total of 45 427 children were examined in Zhenjiang City from 2006 to 2018, and 145 children were positive for E. vermicularis infections, with a mean prevalence of 0.32%. The prevalence of E. vermicularis infections decreased from 1.33% in 2006 to 0.22% in 2018 in children, and the overall prevalence appeared a decline tendency (P < 0.01). A single parasite infection was predominant in soil-transmitted nematode infections (97.46%), and mixed infection of two parasites were mainly detected before 2007. A total of 535 089 person-times received deworming in Zhenjiang City from 2006 to 2018, and the rate of adverse events was 0.002%. Conclusions Currently, the soil-transmitted nematode infection is at a low level in Zhenjiang City, and the overall prevalence of soil-transmitted nematode infections shows a decline tendency year by year. The surveillance and health education of soil-transmitted nematodiasis should be further intensified to consolidate the control achievements.

Objective To investigate the prevalence of parasitic infections in freshwater fish and marine fish from markets in Zhenjiang City, so as to provide the evidene for formulating targeted control measures. Methods Freshwater fish and marine fish were collected from farmers’ markets and supermarkets in Zhenjiang City using a random sampling method. The metacercaria and larvae of parasites were detected using microscopy and the direct dissection method. The prevalence of parasitic infections was analyzed in freshwater fish and marine fish. Results The overall prevalence of parasite infections was 11.21% (208/1 856) in the 1 856 fish (34 species) detected from 2017 to 2020, and the prevalence was 9.81% (117/1 193) in freshwater fish and 13.73% (91/663) in marine fish. Among the 1 193 freshwater fish (23 species), high prevalence of parasitic infections was detected in Gambusia affinis (43.33%), coarse fish (32.93%) and Hemiculter leucisclus (20.20%), and there was no significant difference in the prevalence of parasitic infections among different species of freshwater fish (χ² = 105.66, P < 0.05). Clonorchis sinensis, Metorchis orientalis and other parasite species were detected in freshwater fish, with detection rates of 0.59% (7/1 193), 1.93% (23/1 193) and 7.29% (87/1 193), respectively, and there was a significant difference in the detection of C. sinensis and M. orientalis in freshwater fish (χ² = 8.64, P < 0.05). The prevalence and intensity of larval Anisakis infections were 13.73% (91/663) in marine fish and 3.43 parasites per fish. High prevalence of Anisakis infections was seen in little yellow croaker fish (30.28%) and hairtail (27.50%), and there was a significant difference in the prevalence of parasitic infections in different species of marine fish (χ² = 12.93, P < 0.05). Conclusions Parasitic infections are prevalent in freshwater fish and marine fish from markets of Zhenjiang City, and there is a risk of parasitic infections following consumption of raw or under-cooked freshwater fish and marine fish.

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

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