1.Feasibility Evaluation of using PAX1/JAM3 methylation markers as cervical cancer screening for female self-collected samples
Furong YU ; Jiezhi MA ; Xi ZHOU ; Genlin LI ; Jiaqi PENG ; Ping LI ; Fei ZENG ; Xiaobing XIE ; Weilei DONG
Chinese Journal of Laboratory Medicine 2024;47(4):419-427
Objective:To explore the application value of PAX1/JAM3 methylation detection by cervical self-collected specimen in cervical cancer screening and the management of premenopausal and postmenopausal women.Method:This study is a single center cross-sectional study. From January 2023 to November 2023, cervical self-collected and physician-collected specimens at the colposcopy clinic were detected the PAX1/JAM3 methylation (PAX1 m/JAM3 m) testing. The consistency between self-collected and physician-collected specimens for PAX1 m/JAM3 m detection were compared based on histopathology. In addition, the clinical efficacy of methylation detection with high-risk human papillomavirus (hrHPV), liquid-based cytology (LBC), and their combination for cervical cancer screening were compared in the study. Results:A total of 301 women were recruited to undergo referral colposcopy examination, and statistical analysis was conducted on 272 women with pathological and diagnostic information. Among them, 102 cases (37.5%) were diagnosed as normal cervical tissue or chronic cervicitis, 72 cases (26.4%) were cervical intraepithelial neoplasia grade 1 (CIN1), 43 cases (15.8%) were CIN2, 29 cases (10.7%) were CIN3, and 26 cases (9.6%) were cervical cancer. According to the minimum quantity formula, they were divided into a consistency cohort of 81 participants and a validation cohort of 191 participants. The consistency between cervical self-collected and physician-collected specimens for detecting PAX1 m/JAM3 m. Results from spearman correlation analysis showed a positive correlation between the self-collected and physician-collected results of PAX1 m/JAM3 m detection, and the correlation coefficient R values are 0.858 ( P<0.001) and 0.828 ( P<0.001). The sensitivity and specificity of PAX1 m/JAM3 m detection for diagnosing CIN2 or more severe lesions (CIN2+) were 77.6% [95% confidence interval ( CI) 65.3%-86.4%] and 87.2% (95% CI 80.5%-91.9%), respectively. In clinical performance comparison, the sensitivity of PAX1 m/JAM3 m combined with HPV16/18 detection, 89.7% (95% CI 79.2%-95.2%), was the same as that of hrHPV detection in CIN2+and 96.0% (95% CI 80.4%-99.3%) in CIN3+, which is higher than 92.0% (95% CI 75.0%-97.8%) of hrHPV and 82.6% (95% CI 62.9%-93.0%) of LBC or the combination of sPAX1 m/JAM3 m and LBC low-grade and higher squamous intraepithelial lesion testing [87.0% (95% CI 67.9%-95.5%)]. Conclusions:Self-collected specimens by women for detection of PAX1 and JAM3 methylation as a promising screening tool for cervical cancer has operational and clinical feasibility. The methylation test can optimize the current cervical cancer screening plan, reduce the number of referral women with false positive diagnosis to colposcopy, and is of great significance for reducing fertility protection and preventing missed diagnosis in women of childbearing age.
2.Cloning, expression and purification of fructose-2, 6-bisphosphatase gene CpF2KP in papaya.
Liping ZUO ; Qiuxia ZENG ; Xiaobing ZHAO ; Liyuan YANG ; Liangwei XU ; Juan LAI ; Jingjing YUE
Chinese Journal of Biotechnology 2023;39(2):614-624
Papaya, which is mainly cultivated in the southeastern region of China, is one of the four famous fruits in Lingnan. It is favored by people because of its edible and medicinal value. Fructose-6-phosphate, 2-kinase/fructose-2, 6-bisphosphatase (F2KP) is a unique bifunctional enzyme with a kinase domain and an esterase domain that catalyzes the synthesis and degradation of fructose-2, 6-bisphosphate (Fru-2, 6-P2), an important regulator of glucose metabolism in organisms. In order to study the function of the gene CpF2KP encoding the enzyme in papaya, it is particularly important to obtain the target protein. In this study, the coding sequence (CDS) of CpF2KP, with a full-length of 2 274 bp, was got from the papaya genome. The amplified sequence of full-length CDS was cloned into the vector PGEX-4T-1 which was double digested with EcoR I and BamH I. The amplified sequence was constructed into a prokaryotic expression vector by genetic recombination. After exploring the induction conditions, the results of SDS-PAGE showed that the size of the recombinant GST-CpF2KP protein was about 110 kDa. The optimum IPTG concentration and temperature for CpF2KP induction were 0.5 mmol/L and 28 ℃, respectively. The purified sin[A1] gle target protein was obtained after purifying the induced CpF2KP protein. In addition, the expression level of this gene was detected in different tissues, and showed that the gene was expressed at the highest level in seeds and the lowest in pulp. This study provides an important basis for further revealing the function of CpF2KP protein and studying the involved biological processes of this gene in papaya.
Humans
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Carica/genetics*
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Recombinant Proteins
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Carbohydrate Metabolism
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Cloning, Molecular
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China
3.Difficult and complicated oral ulceration: an expert consensus guideline for diagnosis.
Xin ZENG ; Xin JIN ; Liang ZHONG ; Gang ZHOU ; Ming ZHONG ; Wenmei WANG ; Yuan FAN ; Qing LIU ; Xiangmin QI ; Xiaobing GUAN ; Zhimin YAN ; Xuemin SHEN ; Yingfang WU ; Lijie FAN ; Zhi WANG ; Yuan HE ; Hongxia DAN ; Jiantang YANG ; Hui WANG ; Dongjuan LIU ; Hui FENG ; Kai JIAO ; Qianming CHEN
International Journal of Oral Science 2022;14(1):28-28
The complexity of oral ulcerations poses considerable diagnostic and therapeutic challenges to oral specialists. The expert consensus was conducted to summarize the diagnostic work-up for difficult and complicated oral ulcers, based on factors such as detailed clinical medical history inquiry, histopathological examination, and ulceration-related systemic diseases screening. Not only it can provide a standardized procedure of oral ulceration, but also it can improve the diagnostic efficiency, in order to avoid misdiagnosis and missed diagnosis.
Consensus
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Humans
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Oral Ulcer/therapy*
4.Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (version 2022)
Zhengwei XU ; Dingjun HAO ; Liming CHENG ; Baorong HE ; Bohua CHEN ; Chen CHEN ; Fei CHE ; Jian CHEN ; Qixin CHEN ; Liangjie DU ; Shunwu FAN ; Zhong FANG ; Shiqing FENG ; Yanzheng GAO ; Haishan GUAN ; Zhong GUAN ; Hua JIANG ; Weimin JIANG ; Dianming JIANG ; Jun JIANG ; Yue JIANG ; Lijun HE ; Yuan HE ; Bo LI ; Tao LI ; Jianjun LI ; Xigong LI ; Yijian LIANG ; Bin LIN ; Bin LIU ; Bo LIU ; Yong LIU ; Zhibin LIU ; Xuhua LU ; Chao MA ; Lie QIAN ; Renfu QUAN ; Hongxun SANG ; Haibo SHEN ; Jun SHU ; Honghui SUN ; Tiansheng SUN ; Jun TAN ; Mingxing TANG ; Sheng TAO ; Honglin TENG ; Yun TIAN ; Jiwei TIAN ; Qiang WANG ; Xinwei WANG ; Jianhuang WU ; Peigen XIE ; Weihong XU ; Bin YAN ; Yong YANG ; Guoyong YIN ; Xiaobing YU ; Yuhong ZENG ; Guoqing ZHANG ; Xiaobo ZHANG ; Jie ZHAO ; Yue ZHU
Chinese Journal of Trauma 2022;38(11):961-972
Osteoporotic vertebral compression fracture (OVCF) can lead to lower back pain and may be even accompanied by scoliosis, neurological dysfunction and other complications, which will affect the daily activities and life quality of patients. Vertebral augmentation is an effective treatment method for OVCF, but it cannot correct unbalance of bone metabolism or improve the osteoporotic status, causing complications like lower back pain, limited spinal activities and vertebral refracture. The post-operative systematic and standardized rehabilitation treatments can improve curative effect and therapeutic efficacy of anti-osteoporosis, reduce risk of vertebral refracture, increase patient compliance and improve quality of life. Since there still lack relevant clinical treatment guidelines for postoperative rehabilitation treatments following vertebral augmentation for OVCF, the current treatments are varied with uneven therapeutic effect. In order to standardize the postoperative rehabilitation treatment, the Spine Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized relevant experts to refer to relevant literature and develop the "Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (2022 version)" based on the clinical guidelines published by the American Academy of Orthopedic Surgeons (AAOS) as well as on the principles of scientificity, practicality and advancement. The guideline provided evidence-based recommendations on 10 important issues related to postoperative rehabilitation treatments of OVCF.
5.Progress in research of allergen detection methods
Jie MAO ; Qingqing LU ; Xin ZENG ; Ping LI ; Shengjie SHI ; Jing LI ; Zhenhua ZHU ; Xiaobing XIE ; Qun LU
Chinese Journal of Preventive Medicine 2021;55(1):123-129
Allergic diseases have continued to increase year by year causing serious physical and mental injury to patients, families and individuals. This increase has been driven by conventional environmental and nutritional changes but is also created by the continual introduction of food additives into the diet and novel interior decoration materials into the living space. The causes of allergic diseases are complex and diverse, and the medical laboratory often is not be able to identify the allergic trigger; this creates a difficult environment to identify the appropriate clinical treatment for disease prevention and control. Physicians must be able to identify these triggers to help patients avoid the underlying allergenic cause of their disease. This can only be done by actively knowing a patient′s medical history, identifying the clinical manifestations of hypersensitivity and utilizing confirmatory testing as an important clinical tool in identifying the allergic source.
6.Progress in research of allergen detection methods
Jie MAO ; Qingqing LU ; Xin ZENG ; Ping LI ; Shengjie SHI ; Jing LI ; Zhenhua ZHU ; Xiaobing XIE ; Qun LU
Chinese Journal of Preventive Medicine 2021;55(1):123-129
Allergic diseases have continued to increase year by year causing serious physical and mental injury to patients, families and individuals. This increase has been driven by conventional environmental and nutritional changes but is also created by the continual introduction of food additives into the diet and novel interior decoration materials into the living space. The causes of allergic diseases are complex and diverse, and the medical laboratory often is not be able to identify the allergic trigger; this creates a difficult environment to identify the appropriate clinical treatment for disease prevention and control. Physicians must be able to identify these triggers to help patients avoid the underlying allergenic cause of their disease. This can only be done by actively knowing a patient′s medical history, identifying the clinical manifestations of hypersensitivity and utilizing confirmatory testing as an important clinical tool in identifying the allergic source.
7.Preliminary study of serum 2019-nCoV IgM and IgG antibodies in the diagnosis of COVID-19
Ping LI ; Zhiyong LI ; Silin ZHAO ; Qiong LI ; Yan HU ; Yufeng CHEN ; Fan YI ; Qichen XIE ; Zhaoqiong ZENG ; Changjuan DENG ; Zhanxiang WANG ; Xiaobing XIE
Chinese Journal of Laboratory Medicine 2020;43(4):352-357
Objective:To analyze the clinical value of serum 2019 novel coronavirus (2019-nCoV) immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies in the diagnosis of COVID-19.Methods:A total of 116 patients diagnosed with NCP in the First Affiliated Hospital of Hunan University of Chinese Medicine and the First Affiliated Hospital of Xiamen University were enrolled from January to February 2020 as the disease group. A total of 134 cases, including 84 non-NCP inpatients and 50 healthy individuals served as the control group. Serum samples from all subjects were collected. A fully-automated chemiluminescence immunoassay analyzer was used to detect the concentration of 2019-nCoV IgM and IgG antibodies in serum. The sensitivity and specificity of the 2019-nCoV IgM and IgG antibody single test and combined detection were compared using the χ 2 test. χ 2 test and Wilcoxon′s rank sum test were used to compare the positive rates and concentrations of IgM and IgG antibodies in NCP patients before and after their 2019-nCoV nucleic acid tests turning negative, respectively. The change trend of 2019-nCoV antibody concentration in the process of NCP patients was analyzed by Wilcoxon′s rank sum test. Results:The sensitivity of 2019-nCoV IgG (90.5%, 105/116) was higher than that of 2019-nCoV IgM (75.9%, 88/116), the difference was statistically significant (χ 2=8.91, P<0.05); The specificity of 2019-nCoV IgG (99.3%,133/134) was higher than that of 2019-nCoV IgM (94.0%, 126/134), the difference was statistically significant (χ 2=5.63, P<0.05). The sensitivity (89.7%,87/97) of 2019-nCoV IgM combined with IgG was higher than that of 2019-nCoV IgM, the difference was statistically significant (χ 2=6.89, P<0.05). The specificity (100%, 125/125) of 2019-nCoV IgM combined with IgG was higher than that of 2019-nCoV IgM, the difference was statistically significant (χ 2=7.70, P<0.05). After 2019-nCoV nucleic acid test converted to negative, the positive rate (9/17) and concentration [13.0 (4.9, 24.7) AU/ml] of serum 2019-nCoV IgM antibody were significantly lower than those when the nucleic acid test was positive, positive rate (15/17) and concentration [29.5 (14.0, 61.3) AU/ml], respectively (χ 2=5.10, Z=-3.195, both P<0.05). In the course of NCP, patients′ serum samples were collected from the first day of diagnosis to every three days, three times in total. The first 2019-nCoV IgM and IgG antibody concentrations [19.4 (12.4, 63.7) AU/ml, 105.8 (74.8, 126.1) AU/ml, respectively] were significantly higher than the second concentrations [15.8 (7.1, 40.3)AU/ml, 80.5 (66.7, 105.9) AU/ml], Z were-2.897,-3.179, both P<0.05. Conclusions:2019-nCoV IgG antibody has a good application value in the diagnosis of NCP. The concentration of 2019-nCoV IgM antibody has a certain correlation with the detection of 2019-nCoV nucleic acid. The combination of 2019-nCoV IgM and IgG antibodies with 2019-nCoV nucleic acid test may be the best laboratory index for the diagnosis of NCP at present.
8.Preliminary study of the antibody level in confirmed patients with COVID-19 after discharge
Ge SHEN ; Gang YANG ; Ziyan ZENG ; Yan HU ; Qiong LI ; Zugui LIU ; Huiyuan FU ; Junyu HU ; Pan ZHU ; Juhua HUANG ; Qingqing LU ; Shengjie SHI ; Ying HE ; Xiaobing XIE
Chinese Journal of Preventive Medicine 2020;54(12):1448-1452
Objective:To analyze the antibody levels and dynamic changes in patients infected with 2019-novel coronavirus(2019-nCoV).Methods:The average age of 72 corona virus disease 2019 (COVID-19) patients was (45.53±16.74)years(median age:47 year), including (44.88±17.09) years(median age:46 year) for 38 males and (46.32±16.52)years (median age:46 year) for 34 females in Loudi City, Hunan Province. There is no significant difference in genders between the severe and mild groups (χ2=0.916, P>0.05). There is a significant difference in the age between the severe and mild groups ( F=3.315, P<0.05). The blood samples of 72 discharged patients were collected and the consistence of IgM and IgG antibodies were detected by chemiluminescence method. SPSS25.0 was used for gender, age, case type and antibody analysis of variance, χ 2 test and other analysis. Results:The average time of the serum samples collection of 72 patients was (34.89±9.02)days (median time: 34 days) from onset of COVID-19, and (14.53±8.35) days (median time: 14 days) from discharge. The positive rate of IgM or IgG was 97.22% (70/72), and the positive rate of IgM and IgG was 48.61% (35/72) and 97.22% (70/72) respectively. Serum COVID-19 antibodies were detected in 72 patients from 1st to 40th days after discharge. The average concentration of IgM in 1-7 days, 8-14 days, 15-21 days, 22-28 days, above 29 days were 21.91(7.07-52.84)AU/ml, 14.16(6.19-32.88)AU/ml, 11.36(6.65-42.15)AU/ml, 8.15(3.66-30.12)AU/ml, 2.98(0.46-6.37)AU/ml. There was no significant difference in the time of IgM antibody concentration ( H= 8.439, P>0.05). The average concentrations of IgG in 1-7 days, 8-14 days, 15-21 days, 22-28 days, 29 days and above were 169.90 (92.06-190.91) AU/ml, 163.89 (91.19-208.02) AU/ml, 173.31 (95.06-191.28) AU/ml, 122.84 (103.19-188.34) AU/ml, 101.98 (43.75-175.30) AU/ml, respectively, ( H=2.232, P>0.05). The IgM becomes negative after the 3rd week of discharge and decreases rapidly with time. The IgG concentration higher than IgM during the same period, and keep at high level without any change, and decrease in the fourth week. Among them, 5 cases developed "re-infection" within 1-3 weeks after discharge, and the rate of "re-infection" was 6.94% (5/72 cases). Conclusions:After the COVID-19 patients are discharged from the hospital, the level of antibodies produced varies greatly among individuals, but the overall changes in antibodies have a certain pattern. It is recommended to strengthen the antibody monitoring during hospitalization and after discharge from the hospital to reduce the "re-infection" rate and potential risk of infection.
9.Preliminary study of the antibody level in confirmed patients with COVID-19 after discharge
Ge SHEN ; Gang YANG ; Ziyan ZENG ; Yan HU ; Qiong LI ; Zugui LIU ; Huiyuan FU ; Junyu HU ; Pan ZHU ; Juhua HUANG ; Qingqing LU ; Shengjie SHI ; Ying HE ; Xiaobing XIE
Chinese Journal of Preventive Medicine 2020;54(12):1448-1452
Objective:To analyze the antibody levels and dynamic changes in patients infected with 2019-novel coronavirus(2019-nCoV).Methods:The average age of 72 corona virus disease 2019 (COVID-19) patients was (45.53±16.74)years(median age:47 year), including (44.88±17.09) years(median age:46 year) for 38 males and (46.32±16.52)years (median age:46 year) for 34 females in Loudi City, Hunan Province. There is no significant difference in genders between the severe and mild groups (χ2=0.916, P>0.05). There is a significant difference in the age between the severe and mild groups ( F=3.315, P<0.05). The blood samples of 72 discharged patients were collected and the consistence of IgM and IgG antibodies were detected by chemiluminescence method. SPSS25.0 was used for gender, age, case type and antibody analysis of variance, χ 2 test and other analysis. Results:The average time of the serum samples collection of 72 patients was (34.89±9.02)days (median time: 34 days) from onset of COVID-19, and (14.53±8.35) days (median time: 14 days) from discharge. The positive rate of IgM or IgG was 97.22% (70/72), and the positive rate of IgM and IgG was 48.61% (35/72) and 97.22% (70/72) respectively. Serum COVID-19 antibodies were detected in 72 patients from 1st to 40th days after discharge. The average concentration of IgM in 1-7 days, 8-14 days, 15-21 days, 22-28 days, above 29 days were 21.91(7.07-52.84)AU/ml, 14.16(6.19-32.88)AU/ml, 11.36(6.65-42.15)AU/ml, 8.15(3.66-30.12)AU/ml, 2.98(0.46-6.37)AU/ml. There was no significant difference in the time of IgM antibody concentration ( H= 8.439, P>0.05). The average concentrations of IgG in 1-7 days, 8-14 days, 15-21 days, 22-28 days, 29 days and above were 169.90 (92.06-190.91) AU/ml, 163.89 (91.19-208.02) AU/ml, 173.31 (95.06-191.28) AU/ml, 122.84 (103.19-188.34) AU/ml, 101.98 (43.75-175.30) AU/ml, respectively, ( H=2.232, P>0.05). The IgM becomes negative after the 3rd week of discharge and decreases rapidly with time. The IgG concentration higher than IgM during the same period, and keep at high level without any change, and decrease in the fourth week. Among them, 5 cases developed "re-infection" within 1-3 weeks after discharge, and the rate of "re-infection" was 6.94% (5/72 cases). Conclusions:After the COVID-19 patients are discharged from the hospital, the level of antibodies produced varies greatly among individuals, but the overall changes in antibodies have a certain pattern. It is recommended to strengthen the antibody monitoring during hospitalization and after discharge from the hospital to reduce the "re-infection" rate and potential risk of infection.
10.Differential expression of miRNAs in breast cancer tissues and the effect of Xihuang Pill on the ex-pression of candidate miRNAs in breast cancer cells in vitro
Tao XU ; Ping LI ; Guolian PANG ; Fan YI ; Zhaoqiong ZENG ; Xiaobing XIE
Journal of Chinese Physician 2019;21(7):979-983
Objective To study the differentially expressed microRNAs ( miRNAs) in breast canc-er formalin-fixed, paraffin-embedded ( FFPE) tissue and the effect of Xihuang Pill extract on the expression of candidate miRNAs in breast cancer cell line in vitro. Methods Microarray was used to detect the differ-entially expressed miRNAs in breast cancer tissues, adjacent tissues and cancer tissues with different molec-ular types, and the results of unsupervised cluster analysis were processed by cluster software. Real-time fluorescence quantitative polymerase chain reaction ( qPCR) was used to detect the expression of candidate miRNAs in 106 breast cancer tissues, 22 paracancerous tissues and 66 benigh breast lesions. The extract of Xihuang Pill interfered with T-47D cells and MDA-MB-231 cells, then the expression of candidate miRNAs in those cells were detected by qPCR. Results The microarray results showed that the expression of miR-130b was higher in cancer tissues than in adjacent tissues (P<0. 05). The expression of miR-205 in basal-like cancer tissues with higher malignancy was significantly lower than that in luminal cancer tissues ( P<0. 05). qPCR showed that the expression miR-130b in cancer tissues (t=6. 123, P=0. 000) and adjacent tissues (t= -3. 375, P=0. 001) were significantly higher than that in benigh lesion tissues, and the ex-pression of miR-205 in cancer tissues was significantly lower than that in benigh lesion tissues ( t =-2. 073, P=0. 041). Cell experiment in vitro showed that the expression of miR-130b in T-47D cells de-creased after the intervention of Xihuang Pill extract (t=4. 341, P=0. 012) , while the expression of miR-205 in MDA-MB-231 cells increased after the intervention of Xihuang Pill extract ( t = -3. 266, P =0. 031). Conclusions The increase of miR-130b and the decrease of miR-205 are related to the occur-rence and development of breast cancer. The increase of miR-130b and the decrease of miR-205 may be the one of anti-tumor ways of Xihuang Pill.

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