Objective:To evaluate the value of diagnosis and therapy of cold knife conization with electro-cautery hemostasis by hysteroscope in the treatment of ceNical intraepithelial neoplasia (CIN).Methods :A retrospective analysis of the clinical data was carried out in 193 cases with CIN underwent cold knife coniza-tion with electrocautery hemostasis by hysteroscope from January 2005 to November 2008, and all patients had pathological diagnosis under colposcopic biopsy.Results:The operative time was from 15 to 40 mi-nutes, and the blood loss dunng operation was from 5 to 25 milliliters.The coincidence rate of histopathology before and after conization was 67.88% in 131 cases.9 CIN Ⅲ patients had positive margins after opera-tion, owing to scab break off bleeding of cervical wound was encountered in 18 cases.No infection and cervi-cal adhesion or stenosis occurred.Conclusions :Cold knife conization with electrocautery hemostasis by hyst-eroscope is an effective diagnosis and treatment for CIN.

Background and purpose:miR-101 has been reported to be down-regulated in gastric cancer, colorectal cancer, breast cancer as well as prostate cancer acting as a tumor suppressor gene. However, its function in ovarian cancer is still unknown. The aim of this study was to investigate whether miR-101 can suppress cell growth and invasion of ovarian cancer cells by targeting DNMT3A, so as to reveal molecular mechanism to inhibit ovarian cancer. Methods:Quantitative real-time palymerase chain reaction (qRT-PCR) method was employed to detect the expression of miR-101 in ovarian cancer and cancer adjacent normal ovarian tissues. SKOV3 cells were transfected with miR-101 mimics, and DNMT3A siRNA was transfected as a positive control. Then Western blot was used to detect the expres-sion of DNMT3A protein regulated by miR-101 in SKOV3 cells. The growth and invasion ability of SKOV3 cells were evaluated by MTT and Transwell invasion assays.Results:qRT-PCR showed that miR-101 was down-regulated in ovarian cancer tissues. Western blot showed that the level of DNMT3A protein was inhibited by restored miR-101 or knock-down of DNMT3A in SKOV3 cells. Following transfection of miR-101 mimics or knock-down of DNMT3A for 48, 72 and 96 h respectively, MTT assay showed that theD values were signiifcantly lower than the control group, (P<0.05). After transfection of miR-101 mimics or knock-down of DNMT3A for 36 h, Transwell invasion assay showed that the numbers of cells through the basement membrane was (105±7) and (107±13), respectively, which are signiifcantly different from the control group (213±11), indicating invasion of SKOV3 cells signiifcantly slowed down (P<0.05).Conclusion:miR-101 suppresses cell growth and invasion by targeting DNMT3A in ovarian cancer.

To establish a time-resolved fluorescence immunochromatographic assay for quantitative determination of carbohydrate antigen 19-9 (CA19-9) in serum, we prepared CA19-9 test strips by integrating double-antibody sandwich method and fluorescence immunochromatography technique. Carboxy fluorescent microspheres and nitrocellulose membrane were used as carriers for labeling and coating CA19-9 pairing antibodies. We optimized the process by adjusting the amount of labeling and coating antibody. According to the linear range, lowest detection limit and precision, We evaluated the time-resolved fluorescence immunochromatographic assay of CA19-9. When the amount of labeled antibody was 80 μg for 20 μL fluorescent microspheres, and the concentration of coated antibody on the test line was 1.5 mg/mL, the optimal reaction time was 15 minutes. Assay linear range was 12.5 to 800 U/mL and the minimum detection limit was 6.32 U/mL. The Within-run and between-run coefficient of variation were less than 15%. Average recovery rate was 101%. By detecting 50 clinical samples in parallel with Roche electrochemical luminescence detection kit, correlation coefficient was 0.980 6. The experiment, initially established a fluorescence immunochromatographic detection method to quantitative detection of serum CA19-9, which has a good clinical application prospect.

OBJECTIVE: To investigate the sequences of internal transcribed spacer 2 (ITS2) and cyclooxygenase 1 (COX1) genes of Paragonimus metacercariae in freshwater crabs in Henan Province, identify the species of Paragonimus and evaluate its genetic relationships with Paragonimus isolates from other provinces in China. METHODS: Freshwater crabs were collected from 8 survey sites in Zhengzhou, Luoyang, Pingdingshan, Nanyang and Jiyuan cities of Henan Province from 2016 to 2021, and Paragonimus metacercariae were detected in freshwater crabs. Genomic DNA was extracted from Paragonimus metacercariae, and the ITS2 and COX1 genes were amplified using PCR assay, followed by sequencing of PCR amplification products. The gene sequences were spliced and aligned using the software DNASTAR, and aligned with the sequences of Paragonimus genes in the GenBank. Phylogenetic trees were created using the MEGA6 software with the Neighbor-Joining method based on ITS2 and COX1 gene sequences, with Fasciola hepatica as the outgroup. RESULTS: The detection rates of Paragonimus metacercariae were 6.83% (11/161), 50.82% (31/61), 18.52% (5/26), 8.76% (12/137), 14.29% (9/63), 17.76% (19/105), 18.50% (32/173) and 42.71% (41/96) in freshwater crabs from 8 survey sites in Zhengzhou, Luoyang, Pingdingshan, Nanyang and Jiyuan cities of Henan Province, with a mean detection rate of 19.46% (160/822), and a mean infection intensity of 0.57 metacercariae/g. The amplified ITS2 and COX1 gene fragments of Paragonimus were approximately 500 bp and 450 bp in lengths, respectively. The ITS2 gene sequences of Paragonimus metacercariae from 8 survey sites of Henan Province showed the highest homology (99.8% to 100.0%) with the gene sequence of P. skrjabini (GenBank accession number: MW960209.1), and phylogenetic analysis showed that the Paragonimus in this study was clustered into the same clade with P. skrjabini from Sichuan Province (GenBank accession number: AY618747.1), Guangxi Zhuang Autonomous Region (GenBank accession number: AY618729.1) and Hubei Province (GenBank accession number: AY618751.1), and P. miyazaki from Fujian Province (GenBank accession number: AY618741.1) and Japan (GenBank accession number: AB713405.1). The COX1 gene sequences of Paragonimus metacercariae from 8 survey sites of Henan Province showed the highest homology (90.0% to 100.0%) with the gene sequence of P. skrjabini (GenBank accession number: AY618798.1), and phylogenetic analysis showed that the Paragonimus in this study was clustered into the same clade with all P. skrjabini and clustered into the same sub-clade with P. skrjabini from Hubei Province (GenBank accession numbers: AY618782.1 and AY618764.1). CONCLUSIONS Paragonimus species from freshwater crabs in Henan Province were all characterized as P. skrjabini, and the ITS2 and COX1 gene sequences had the highest homology to those of P. skrjabini from Hubei Province. The results provide insights into study of Paragonimus in Henan Province and China.
Animals ; Paragonimus/genetics* ; Brachyura/genetics* ; Cyclooxygenase 1/genetics* ; Phylogeny ; China/epidemiology* ; Sequence Analysis, DNA ; Paragonimiasis

Objective To analyze the epidemiological characteristics of echinococcosis cases reported in the National Notifiable Disease Report System in Henan Province from 2010 to 2011, so as to provide insights into for echinococcosis control and surveillance. Methods The data pertaining to reported echinococcosis cases in Henan Province from 2010 to 2021 were retrieved from the National Notifiable Disease Report System, and a descriptive epidemiological analysis was performed using the software SPSS 22.0. Results A total of 150 echinococcosis cases were reported in Henan Province from 2010 to 2021, including 88 confirmed cases (58.67%) and 62 clinically diagnosed cases (41.33%), 77 cases reported by Henan Province (51.33%) and 73 cases reported by other provinces (48.67%). Echinococcosis cases were reported in each month, with 8 to 21 cases reported in each month, and the number of reported echinococcosis cases appeared no remarkable temporal changes. The echinococcosis cases were reported across 18 cities of Henan Province, with the highest number of cases reported in Zhoukou (17.33%) and Nanyang cities (17.33%) and the lowest number reported in Sanmenxia City (0.67%). The reported echinococcosis cases had a male to female ratio of 1.17:1, and the cases were found at each age group, with the the highest number of cases seen at ages of 20 to 59 years (73.33%). Farmer was the predominant occupation (63.33%), followed by housekeepers and the unemployed (12.67%). Of all reported echinococcosis cases, there were 25 local cases (16.67%) and 125 imported cases (83.33%), 144 cases reported by medical institutions (96.00%) and 6 cases reported by centers for disease control and prevention (4.00%). Conclusions Although imported echinococcosis cases were the predominant source of echinococcosis cases reported in the National Notifiable Disease Report System in Henan Province from 2010 to 2021, there were still sporadic local cases, and the emergence of local sources of infection cannot be excluded. Further expanded field surveys and surveillance of echinococcosis are required.

[Abstract] Objective: To investigate the effect of Beclin1 knockdown on cisplatin resistance in ovarian cancer A2780 cells and its related mechanisms. Methods: The mRNA and protein expressions of Beclin1 in A2780 cells and drug resistant A2780/DDP cells were determined by qPCR and Western blotting. After transfection with Beclin1 siRNA, the sensitivity of A2780/DDP cells to cisplatin was detected by MTT assay; Cell clone formation and apoptosis were detected by the Colony formation assay and Flow cytometry assay, respectively; cell autophagy was monitored by monodansylcadaverin (MDC) staining. Furthermore, the protein levels of cell autophagy related proteins, lysosomal associated membrane protein Lamp-2 and Cathepsin B were detected by Western blotting. Results: The mRNA and protein expression levels of Beclin1 in cisplatin-resistant A2780/DDP cells were significantly higher than those in A2780 cells (all P<0.05). The expression of Beclin1 was significantly increased in A2780 cells after treated with cisplatin (P<0.05). Beclin1 knockdown promoted cisplatin induced apoptosis of A2780/DDP cells (P<0.05), inhibited autophagy and cell colony formation (all P<0.05), and increased cell sensitivity to cisplatin (P<0.05). Meanwhile, Western blotting showed that Beclin1 knockdown increased the protein levels of cleaved-caspase 3 and Cathepsin B in A2780/DDP cells, while down-regulated the protein expressions of Atg3, Atg7, LC3Ⅱ/Ⅰand Lamp-2 (all P<0.05). Conclusion: Beclin1 knockdown can improve the sensitivity of A2780/DDP cells to cisplatin, and the mechanism may be related to the inhibition of protective autophagy of cells by regulating the expressions of autophagy related proteins, and the regulation of lysosomes, thus further promoting cisplatin-induced apoptosis of drug-resistant cells.

Objective To investigate the changes in the awareness rate of Taenia solium taeniasis and cysticercosis control knowledge among medical professionals before and after training in Fangcheng County, a disease-elimination pilot area of Henan Province, so as to evaluate the effectiveness of the training. Methods Three townships in Fangcheng County were randomly selected as the study townships, including Dushu, Bowang and Yangji townships, while Erlangmiao, Yanglou and Xiaoshidian townships in the county were randomly selected as the control townships. The grassroots medical professionals in the study townships were given once training on T. solium taeniasis and cysticercosis control knowledge each year from 2016 to 2020, while those in the control townships were given no interventions. All village-level doctors and a part of township-level public health professionals were sampled from the study and control townships as intervention and control groups. The baseline and final assessments of the awareness of T. solium taeniasis and cysticercosis control knowledge were performed using questionnaire survey in intervention and control groups in 2016 and 2020, and the awareness of T. solium taeniasis and cysticercosis control knowledge was compared between the two groups. Results A total of 663 medical professionals were investigated in Fangcheng County from 2016 to 2020, including 474 participants in the intervention group and 189 participants in the control group. Results from the 2016 baseline survey showed that the awareness rate of T. solium taeniasis and cysticercosis control knowledge was 28.83% (47/163) among grassroots medical professionals in Fangcheng County, and there were no significant differences in the awareness between the intervention (32.47%, 25/77) and control groups (25.58%, 22/86) (χ² = 0.939, P > 0.05), between men (30.50%, 43/141) and women (18.18%, 4/22) (χ² = 1.406, P > 0.05) or between village- (31.39%, 43/137) and township-level medical professionals (15.38%, 4/26) (χ² = 2.727, P > 0.05), while significant differences were found in the awareness rate of T. solium taeniasis and cysticercosis control knowledge among medical professionals in terms of education levels (χ² = 8.190, P < 0.05) and duration of working experiences (χ² = 12.617, P < 0.05), and the awareness rate of T. solium taeniasis and cysticercosis control knowledge increased with education levels among medical professionals (χ² = 6.768, P < 0.05). Only 5.52% (9/163) of the medical professionals had a history of diagnosis and therapy of T. solium taeniasis or cysticercosis, and only 1.23% (2/163) received training on T. solium taeniasis and cysticercosis control knowledge during the past 5 years. Results from the 2020 questionnaire survey showed a higher awareness rate of T. solium taeniasis and cysticercosis control knowledge among medical professionals in the intervention group (93.55%, 116/124) than in the control group (46.60%, 48/103) (χ² = 61.845, P < 0.05), and no significant differences were seen in the awareness rate of T. solium taeniasis and cysticercosis control knowledge among medical professionals in terms of gender, level of medical professionals, duration of working experiences or history of diagnosis/therapy of T. solium taeniasis and cysticercosis in the intervention group (χ² = 1.089, 0.140, 0.081 and 0.453, all P values > 0.05), while there was a significant difference in the awareness rate among medical professionals with different education levels (χ² = 36.338, P < 0.05). In addition, the awareness rate of T. solium taeniasis and cysticercosis control knowledge significantly increased among medical professionals with various chracteristics in 2020 than in 2016. Conclusions In the low-prevalence areas of T. solium taeniasis and cysticercosis, long-term and persistent training may improve the awareness of T. solium taeniasis and cysticercosis control knowledge among grassroots medical professionals, which facilitates the timely identification of T. solium taeniasis and cysticercosis and the establishment of a sensitive disease surveillance system.

OBJECTIVE: To establish an HPLC method for simultaneous determination of 15 monoterpene glycosides in Radix Paeoniae Alba and seed cake of peony. METHODS: The separation was performed on an Agilent Zorbax SB C18-Aq column, using acetonitrile (A) and potassium dihydrogen phosphate solution (B) (pH 2.8) as the mobile phase by gradient elution. The elution program was as follows: 0 min(A, 9%)→8 min(A, 9%)→10 min(A, 15%)→25 min(A, 20%)→42 min(A, 35%)→50 min(A, 35%). The flow rate was 1.0 mL•min-1. The detection wavelength was maintained at 260 nm. The column temperature was set at 30 ℃. RESULTS: The linear range was 5.1-81.6 μg•mL-1 for pyrindylpaeoniflorin, 12.95-207.2 μg•mL-1 for mudanpioside F, 6.2-99.2 μg•mL-1 for oxyalbiflorin, 12.2-195.2 μg•mL-1 for oxypaeoniflorin, 8.0-128.0 μg•mL-1 for 10-hydroxypaeoniflorin, 5.5-88.0 μg•mL-1 for albiflorin, 6.0-96.0 μg•mL-1 for paeoniflorin, 6.0-96.0 μg•mL-1 for oxypaeonidanin, 5.6-89.6 μg•mL-1 for 4-O-methyl-oxypaeoniflorin, 4.7-75.2 μg•mL-1 for galloylpaeoniflorin, 5.4-86.4 μg•mL-1 for 4-O-methyl-paeoniflorin, 5.0-80.0 μg•mL-1 foralbiflorin R1, 5.7-91.2 μg•mL-1 for paeonidanin, 5.4-86.4 μg•mL-1 for benzoyloxypaeoniflorin and 6.7-107.2 μg•mL-1 for benzoylpaeoniflorin. The average recoveries of the 15 monoterpene glycosides, were between 96.8%-105.6% and the RSD values were 1.05%-3.41%. CONCLUSION: The method is simple, accurate and can be used for the quality control of peony.

Objective: To establish an HPLC method for simultaneous determination of 15 monoterpene glycosides pyrindyl- paeoniflorin, mudanpioside F, oxyalbiflorin, oxypaeoniflorin, 10-hydroxypaeoniflorin, albiflorin, paeoniflorin, oxypaeonidanin, 4-O- methyl-oxypaeoniflorin, galloylpaeoniflorin, 4-O-methyl-paeoniflorin, albiflorin R1, paeonidanin, benzoyloxypaeoniflorin, and benzoylpaeoniflorin in Paeoniae Rubra Radix Formula Granule, in order to compare the quality of Paeoniae Rubia Radix Formula Granule from different manufacturers and provide the basis for the establishment of a unified quality control method. Methods The separation was performed on an Agilent Zorbax SB-Aq C18 column (250 mm × 4.6 mm, 5 µm), using acetonitrile and potassium dihydrogen phosphate solution (pH 2.8) as the mobile phase at the flow rate of 1.0 mL/min for a gradient elution. The detection wavelength was set at 260 nm. Results: Paeoniflorin, albiflorin, and oxypaeoniflorin were the three main monoterpene glycosides with the highest content in the Paeoniae Rubra Radix Formula Granule. There were extremely significant difference among the contents of 15 monoterpene glycosides in Paeoniae Rubra Radix Formula Granule produced by different manufacturers. The sample of CSPFKL-KRT was remarkable for the highest content of paeoniflorin and oxypaeoniflorin (73.214 mg and 16.935 mg per gram sample, respectively) and the lowest content of albiflorin among all samples (2.343 mg per gram sample), while sample CSPFKL-XLS was remarkable for the lowest content of paeoniflorin and oxypaeoniflorin (26.327 mg and 4.165 mg per gram sample, respectively) and the highest content of albiflorin among all samples (18.893 mg per gram sample). Conclusion: There were extremely significant difference among the contents of main monoterpene glycosides in Paeoniae Rubra Radix Formula Granule produced by different manufacturers, which may affect the clinical use. The establishment of a unified quality standard plays an important role in the quality control of Paeoniae Rubia Radix Formula Granule.

Objective To evaluate the diagnostic efficiency of four anti-cysticercus IgG, IgG4 or IgM antibody test kits (enzyme-linked immunosorbent assay, ELISA) by different manufacturers, so as to provide insights into the epidemiological investigation and clinical detection of cysticercosis. Methods Forty serum samples from cerebral cysticercosis patients, 100 serum samples from healthy volunteers, 30 serum samples from paragonimiasis skrjabini patients, 17 serum samples from cystic echinococcosis and 19 serum samples from subcutaneous or cerebral sparganosis patients were collected and detected using anti-cysticercus IgG, IgG4 or IgM antibody test kits (brand A) and the anti-cysticercus IgG antibody test kit (brand B). The sensitivity, specificity and false negative rate of the four kits for detection of cysticercosis were estimated. Results The anti-cysticercus IgG, IgG4 or IgM antibody test kits (brand A) showed 95.00% (38/40), 87.50% (35/40), 7.50% (3/40) sensitivities and 98.00% (98/100), 100.00% (100/100) and 100.00% (100/100) for detection of cysticercosis, while the anti-cysticercus IgG antibody test kit (brand B) presented a 75.00% (30/40) sensitivity and 100.00% (100/100) specificity for detection of cysticercosis. The sensitivity for detection of cysticercosis was significantly higher by the anti-cysticercus IgG antibody test kit (brand A) than by the anti-cysticercus IgG antibody test kit (brand B) (χ² = 6.28, P < 0.05); however, no significant difference was seen in the specificity by two kits (χ² = 2.01, P > 0.05). The four ELISA kits showed overall false positive rates of 37.88% (25/66), 22.73% (15/66), 62.12% (41/66) and 15.15% (10/66) for detection of paragonimiasis, echinococcosis and sparganosis (χ² = 37.61, P < 0.05), and the anti-cysticercus IgG antibody test kit (brand A) presented the highest overall false positive rate for detection of paragonimiasis, echinococcosis and sparganosis (χ² = 7.56, P’ < 0.008), while a higher overall false positive rate was seen for detection of paragonimiasis, echinococcosis and sparganosis by the anti-cysticercus IgG antibody test kit (brand A) than by the anti-cysticercus IgG antibody test kit (brand B) (χ² = 8.75, P’ < 0.008). The four ELISA kits showed false positive rates of 40.00% (12/30), 16.67% (5/30), 76.67% (23/30) and 13.33% (4/30) for detection of paragonimiasis (χ2 = 32.88, P < 0.05) and 21.05% (4/19), 26.32% (5/19), 73.68% (14/19) and 15.79% (3/19) for detection of sparganosis (χ² = 19.97, P < 0.05), and the highest false positive rates were found by the anti-cysticercus IgM antibody test kit (brand A) for detection of paragonimiasis and sparganosis (all P’ < 0.008). However, the four ELISA kits showed comparable false positive rates of 52.94% (9/17), 29.41% (5/17), 23.53% (4/17) and 17.65% (3/17) for detection of echinococcosis (χ² = 8.24, P > 0.05). In addition, the anti-cysticercus IgM anti-body test kit (brand A) showed false positive rates of 76.67% (23/30), 23.53% (4/17) and 73.68% (14/19) for detection of paragonimiasis, echinococcosis and sparganosis (χ² = 14.537, P < 0.05), with the lowest false positive rate seen for detection of echinococcosis (χ² = 14.537, P’ < 0.014), while no significant differences were seen in the false positive rate for detection of paragonimiasis, echinococcosis and sparganosis by other three ELISA kits (all P > 0.05). Conclusions The four anti-cysticercus IgG, IgG4 or IgM antibody test kits exhibit various efficiencies for serodiagnosis of cysticercosis. The anti-cysticercus IgG antibody test kit (brand A) has a high sensitivity for serodiagnosis of cysticercosis; however, it still needs to solve the problems of cross-reaction with other parasitic diseases and stability.