1.Characteristics of cervical lesions in 1 080 patients with abnormal colposcopic biopsies of TCT and HPV double screening in Shihezi region
Yangyang RUAN ; Yan WEI ; Jishuai HOU ; Rui BAI ; Xiaohan XU ; Ping YANG
Journal of Chinese Physician 2022;24(1):29-34
Objective:To explore the characteristics of cervical lesions in female patients in the Shihezi region of Xinjiang in the past two years and provide guidance for clinical work and local cervical cancer screening.Methods:The clinicopathological data and characteristics of 1 080 patients with abnormal double screening of human papilloma virus (HPV) and cervical exfoliative cell test (TCT) and vaginal mirror cervical tissue biopsy in the gynecology department of the First Affiliated Hospital of Medical College of Shihezi University from May 2017 to March 2019 were analyzed retrospectively. The influencing factors of cervical lesions were analyzed by logistic regression.Results:(1) Uneducated [odds ratio ( OR)=2.267], irregular vaginal bleeding ( OR=3.275) and number of sexual partners ≥3 ( OR=3.052) were independent risk factors for cervical lesions. (2) Cervical intraepithelial neoplasia (CIN) was detected most frequently in the 30-39 age group (57.54%), and cervical cancer was detected most frequently in the ≥60 age group (14.15%), followed by the 40-49 age group (2.20%). (3) The proportion of HPV positive and the positive rate of high-risk HPV16/18 in the 40-49 age group were the highest, 33.18% and 39.38% respectively. There was no statistically significant difference in HPV viral load between the different age groups ( P>0.05). (4) The detection rates of CIN3 and cervical cancer were higher in the HPV16/18-positive group than in the HPV-positive and TCT≥atypical squamous epithelial cells of undermined significance (ASC-US) group (18.34% vs 11.33%, 4.30% vs 1.82%), with statistically significant difference (all P<0.05), respectively. The detection rate of CIN2 was higher in the high-load group (28.44%) than in the medium-load group (25.32%) and the low load group (15.79%). (5) The detection rate of CIN3 in the HPV-positive TCT for the ASC-US populations was significantly higher in the HPV16/18-positive group than in the other subtypes of HPV-positive group (21.43% vs 8.33%, P<0.05). Conclusions:During cervical cancer screening in Shihezi region, we should strengthen the publicity of elderly women and uneducated people. CIN and cervical carcinoma in Shihezi region are closely associated with high-risk HPV infection, especially HPV16/18 infection. HPV-positive age is mainly concentrated in the age group of 40-49 years, and the detection rate of CIN and cervical cancer is the highest in the age group of 30-39 years and ≥60 years, respectively. The detection rate of ≥CIN2 in HPV16/18 fraction and high viral load population is higher than that of other HPV subtypes positive and low to medium load populations. How to effectively shunt HPV positive TCT is an important problem in ASC-US population, which needs further research.
2.Lipopolysaccharide inhibits lipophagy in HepG2 cells via activating mTOR pathway.
Zheng-Ping HOU ; Yan-Ping LI ; Lei ZHAO ; Ya-Xi CHEN ; Xiong-Zhong RUAN
Acta Physiologica Sinica 2021;73(5):813-820
This study aimed to investigate the effect of lipopolysaccharide (LPS) on lipophagy in hepatocytes and the underlying mechanism. Human hepatoma cell line HepG2 was cultured in vitro, treated with 0.1 mmol/L palmitic acid (PA), and then divided into control group (0 μg/mL LPS), LPS group (10 μg/mL LPS), LPS+DMSO group and LPS+RAPA (rapamycin, 10 μmol/L) group. Lipid accumulation in hepatocytes was observed by oil red O staining. The autophagic flux of the cells was assessed using confocal laser scanning microscope after being transfected with autophagy double-labeled adenovirus (mRFP-GFP-LC3). The level of intracellular lipophagy was visualized by the colocalization of lipid droplets (BODIPY 493/503 staining) and lysosomes (lysosome marker, lysosomal associated membrane protein 1, LAMP1). The expression levels of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), ribosome protein subunit 6 kinase 1 (S6K1), p-S6K1, LC3II/I and P62 protein were examined by Western blot. The results showed that the number of red lipid droplets stained with oil red O was significantly increased in LPS group compared with that in control group (P < 0.001). Moreover, in LPS group, the number of autophagosomes was increased, while the number of autophagolysosomes and the colocalization rate of LAMP1 and BODIPY were significantly decreased (P < 0.05). Meanwhile, the ratios of p-mTOR/mTOR and p-S6K1/S6K1, the ratio of LC3II/LC3I and the protein expression of P62 were significantly increased (P < 0.05) in LPS group. Furthermore, compared with LPS+DMSO group, RAPA treatment obviously reduced the number of lipid droplets and autophagosomes, and raised the number of autophagolysosomes and the colocalization rate of LAMP1 and BODIPY (P < 0.05). In conclusion, the results demonstrate that LPS inhibits lipophagy in HepG2 cells via activating mTOR signaling pathway, thereby aggravating intracellular lipid accumulation.
Autophagy
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Hep G2 Cells
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Humans
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Lipopolysaccharides
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Palmitic Acid
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Signal Transduction
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TOR Serine-Threonine Kinases
3.Effect of Protein Kinase A Activation on Aggregation Function of Platelets.
Meng-Xiao JIANG ; Jun LIU ; Kang-Xi ZHOU ; Hong-Lei YE ; Ren-Ping HU ; Rong YAN ; Chang-Geng RUAN ; Ke-Sheng DAI
Journal of Experimental Hematology 2020;28(3):899-903
OBJECTIVE:
To investigate the effect of protein kinase A (PKA) activation on aggregation funetion of platelets in vitro.
METHODS:
The peripheral blood of healthy adults were collected, and the washed platelets were gained from collected peripheral blood. The washed platelets were treated with PKA activator Forskolin, then the platelet aggregation was induced by using Ristocetin, Thrombin, Collagen and ADP respectively, the platelet aggregation level was detected by the platelet aggregator.
RESULTS:
Compared with the controls, 5 μmol/L forskolin significantly inhibited ADP and collagen-induced platelet aggregation (P<0.001), and showed mild inhibiting effect on Thrombin-induced platelet aggregation (P<0.05). 2.5-10 μmol/L forskolin significantly inhibited ADP and Collagen -induced platelet aggregation (P<0.001); but not showed significantly inhibitory effects on Ristocetin-induced platelet aggregation (P>0.05).
CONCLUSION
PKA activation inhibits agonists-induced platelet aggregation.
Blood Platelets
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Cyclic AMP-Dependent Protein Kinases
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Humans
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Platelet Aggregation
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Platelet Aggregation Inhibitors
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Ristocetin
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Thrombin
4."Medium and Long-term Plan for the Prevention and Control of Chronic Non-communicable Diseases in Shanghai(2018-2030)" in the perspective of public health
Qun-di YANG ; Ping-ping BAO ; Yang ZHENG ; Min-na CHENG ; Ye RUAN ; Chun-fang WANG ; Yan SHI
Shanghai Journal of Preventive Medicine 2020;32(6):487-
The "Medium and Long-term Plan for the Prevention and Control of Chronic Non-communicable Diseases in Shanghai (2018-2030)" was officially released in August 2018.From the perspective of public health, this paper analyzes the background of the plan from the epidemic situation, response and challenges Shanghai City is facing, expounds the comprehensive prevention and control system of chronic diseases including four functional systems, and explains the key preventive and control measures on the different stages of chronic diseases, comparing the evaluation indicators with those of the national plan.This paper will help to better understand the new blueprint for the prevention and control of chronic diseases in Shanghai in the next ten years.
5. Identification and characterization of a novel group of natural antisense transcripts from RNA1.2 gene locus of human cytomegalovirus
Bing-Yang LIU ; Yan-Ping MA ; Ying QI ; Yu-Jing HUANG ; Zhong-Yang LIU ; Meng-Qing LU ; Qiang RUAN
Chinese Medical Journal 2019;132(13):1591-1598
Background:
Natural anti-sense transcripts (NATs), which are transcribed from the complementary DNA strand of annotated genes, exert regulatory function of gene expression. Increasing studies recognized anti-sense transcription widespread throughout human cytomegalovirus (HCMV) genome, whereas the anti-sense transcription of RNA1.2 gene locus has never been investigated. In this study, the transcription of the RNA1.2 anti-sense strand was investigated in clinically isolated HCMV strain.
Methods:
Strand-specific high-through RNA-sequencing (RNA-seq) was performed to find possible anti-sense transcripts (ASTs). For analyzing and visualization of RNA-seq data sets, Integrative Genomics Viewer software was applied. To confirm these possibilities, Northern blotting and rapid amplification of cDNA ends (RACE) were used.
Results:
Transcription of the opposite strand of RNA1.2 gene locus was detected by RNA-sequencing using RNAs extracted from human embryonic lung fibroblasts infected with HCMV clinical isolate HAN. At least three HCMV NATs, named RNA1.2 AST 1, RNA1.2 AST2, and RNA1.2 AST3, were characterized by Northern blotting and RACE analyses. These RNA1.2 ASTs orientated from the complementary strand of RNA1.2 locus during the late phase of HCMV infection. The 5′- and 3′-termini of these transcripts were located within the opposite sequence of the predicted RNA1.2 gene.
Conclusion
A cluster of novel NATs was transcribed from the opposite sequence of the HCMV RNA1.2 gene region.
6.Relationships of Inflammatory Factors and Risk Factors with Different Target Organ Damage in Essential Hypertension Patients.
Chun-Lin LAI ; Jin-Ping XING ; Xiao-Hong LIU ; Jie QI ; Jian-Qiang ZHAO ; You-Rui JI ; Wu-Xiao YANG ; Pu-Juan YAN ; Chun-Yan LUO ; Lu-Fang RUAN
Chinese Medical Journal 2017;130(11):1296-1302
BACKGROUNDAtherosclerosis (AS) is an inflammatory disease. Inflammation was considered to play a role in the whole process of AS. This study aimed to analyze the relationships of inflammatory factors and risk factors with different target organ damages (TOD) in essential hypertension (EH) patients and to explore its clinical significance.
METHODSA total of 294 EH patients were selected and divided into four groups according to their conditions of TOD. Forty-eight healthy subjects were selected as control. The clinical biochemical parameters, serum amyloid A, serum tryptase, and lipoprotein-associated phospholipase A2 (Lp-PLA2) in each group were detected, and the related risk factors were also statistically analyzed.
RESULTSFibrinogen (Fbg) was the most significant independent risk factor in acute coronary syndrome (ACS) group (odds ratio [OR]: 22.242, 95% confidence interval [CI]: 6.458-76.609, P< 0.001) with the largest absolute value of the standardized partial regression coefficient B' (b': 1.079). Lp-PLA2 was the most significant independent risk factor in stroke group (OR: 13.699, 95% CI: 5.236-35.837, P< 0.001) with b' = 0.708. Uric acid (UA) was the most significant independent risk factor in renal damage group (OR: 15.307, 95% CI: 4.022-58.250, P< 0.001) with b' = 1.026.
CONCLUSIONSFbg, Lp-PLA2, and UA are the strongest independent risk factors toward the occurrence of ACS, ischemic stroke, and renal damage in EH patients, thus exhibiting the greatest impacts on the occurrence of ACS, ischemic stroke, and renal damage in EH patients, respectively.
1-Alkyl-2-acetylglycerophosphocholine Esterase ; Aged ; Antihypertensive Agents ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Essential Hypertension ; blood ; complications ; drug therapy ; physiopathology ; Female ; Humans ; Kidney Diseases ; blood ; etiology ; physiopathology ; Logistic Models ; Male ; Middle Aged ; Renal Insufficiency, Chronic ; blood ; etiology ; physiopathology ; Risk Factors ; Serum Amyloid A Protein ; metabolism ; Stroke ; blood ; etiology ; physiopathology ; Tryptases ; blood
7.Human mononuclear macrophages and neutrophils: differences in killing ability and mechanism against Leptospira interrogans
Ping RUAN ; Xu'ai LIN ; Yingying ZHANG ; Haili ZHOU ; Jie YAN ; Xu CHEN ;
Chinese Journal of Microbiology and Immunology 2017;37(6):412-417
Objective To understand the differences in killing ability and mechanism of human mononuclear macrophages and neutrophils against Leptospira interrogans.Methods Human THP-1 and HL-60 cell lines were respectively pretreated with PMA (phorbol 12-myristate 13-acetate) and ATRA (all-trans retinoic acid) to induce their differentiation into macrophages and neutrophils.Confocal microscopy was used to detect the changes in total ROS (reactive oxygen species) and NO (nitric oxide) levels as well as free Ca2+ concentration ([Ca2+]i) in THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils after infection with Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai.Fluorospectrophotometry was applied to analyze the differences in killing ability between THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils against intracellular leptospires before and after treatment with total ROS and NO inhibitors and intracellular free Ca2+ chelator.Results The total ROS and NO levels and [Ca2+]i in THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils were significantly increased after infection with the spirochete (P<0.05).Moreover,the total ROS and NO levels and [Ca2+]i in the former were significantly higher than those in the latter (P<0.05).The THP-1 monocyte-derived macrophages had stronger killing ability against intracellular leptospires than the HL-60 cell-derived neutrophils (P<0.05).Inhibiting total intracellular ROS,NO or free Ca2+ could result in decreased killing ability of THP-1 monocyte-derived macrophages against intracellular leptospires,but not affect the killing ability of HL-60 cell-derived neutrophils.Conclusion Mononuclear macrophages rather than neutrophils act as the main phagocytes eliminating Leptospira interrogans.High levels of total intracellular ROS,NO and free Ca2+ are closely associated with the ability of mononuclear macrophages to kill Leptospira interrogans.
8.Relationships of Inflammatory Factors and Risk Factors with Different Target Organ Damage in Essential Hypertension Patients
Lai CHUN?LIN ; Xing JIN?PING ; Liu XIAO?HONG ; Qi JIE ; Zhao JIAN?QIANG ; Ji YOU?RUI ; Yang WU?XIAO ; Yan PU?JUAN ; Luo CHUN?YAN ; Ruan LU?FANG
Chinese Medical Journal 2017;(11):1296-1302
Background: Atherosclerosis (AS) is an inflammatory disease. Inflammation was considered to play a role in the whole process of AS. This study aimed to analyze the relationships of inflammatory factors and risk factors with different target organ damages (TOD) in essential hypertension (EH) patients and to explore its clinical significance. Methods: A total of 294 EH patients were selected and divided into four groups according to their conditions of TOD. Forty?eight healthy subjects were selected as control. The clinical biochemical parameters, serum amyloid A, serum tryptase, and lipoprotein?associated phospholipase A2 (Lp?PLA2) in each group were detected, and the related risk factors were also statistically analyzed. Results: Fibrinogen (Fbg) was the most significant independent risk factor in acute coronary syndrome (ACS) group (odds ratio [OR]:22.242, 95% confidence interval [CI]: 6.458–76.609, P < 0.001) with the largest absolute value of the standardized partial regression coefficient B''(b'': 1.079). Lp?PLA2 was the most significant independent risk factor in stroke group (OR: 13.699, 95% CI: 5.236–35.837, P < 0.001) with b'' = 0.708. Uric acid (UA) was the most significant independent risk factor in renal damage group (OR: 15.307, 95%CI: 4.022–58.250, P < 0.001) with b'' = 1.026. Conclusions: Fbg, Lp?PLA2, and UA are the strongest independent risk factors toward the occurrence of ACS, ischemic stroke, and renal damage in EH patients, thus exhibiting the greatest impacts on the occurrence of ACS, ischemic stroke, and renal damage in EH patients, respectively.
9.Restrictive Cardiomyopathy Resulting from a Troponin I Type 3 Mutation in a Chinese Family.
Yan-Ping RUAN ; Chao-Xia LU ; Xiao-Yi ZHAO ; Rui-Juan LIANG ; Hui LIAN ; Michael ROUTLEDGE ; Wei WU ; Xue ZHANG ; Zhong-Jie FAN
Chinese Medical Sciences Journal 2016;31(1):1-7
Objective To identify the pathogenic variant responsible for restrictive cardiomyopathy (RCM) in a Chinese family.Methods Next generation sequencing was used for detecting the mutation and Results verified by sequencing. We used restriction enzyme digestion to test the mutation in the family members and 200 unrelated normal subjects without any cardiac inherited diseases when the mutation was identified.Results Five individuals died from cardiac diseases, two of whom suffered from sudden cardiac death. Two individuals have suffered from chronic cardiac disorders. Mutation analysis revealed a novel missense mutation in exon 7 of troponin I type 3 (TNNI3), resulting in substitution of serine (S) with proline (P) at amino acid position 150, which cosegregated with the disease in the family, which is predicted to be probably damaging using PolyPhen-2. The mutation was not detected in the 200 unrelated subjects we tested.Conclusion Using next generation sequencing, which has very recently been shown to be successful in identifying novel causative mutations of rare Mendelian disorders, we found a novel mutation of TNNI3 in a Chinese family with RCM.
10.Identification and immunogenicity analysis of predominant T-B combined antigenic epitopes on the ;outer membrane protein Loa22 of Leptospira interrogans strains
Ping RUAN ; Jinfang ZHAO ; Yang LI ; Jie YAN ; Weilin HU
Chinese Journal of Microbiology and Immunology 2015;(4):292-298
Objective To screen and identify the predominant T-and B-cell ( T-B) combined an-tigenic epitopes on the outer membrane protein Loa22 of pathogenic Leptospira interrogans ( L.interrogans) stains and to further analyze their immunogenicity.Methods PCR analysis was used to detect loa22 gene in L.interrogans strains belonging to eight different serogroups or serovars prevalent in China.The PCR prod-ucts were sequenced after T-A cloning.A prokaryotic expression system for loa22 gene of L.interrogans sero-group Icterohaemorrhagiae serovar Lai strain Lai was constructed.The expressed recombinant protein rLoa22 was extracted by Ni-NTA affinity chromatography.The rabbit anti-rLoa22 serum samples and IgG were pre-pared.The T-B combined antigenic epitopes on Loa22 protein were predicted by using professional bioinfor-matic softwares.Phage display in combination with Western blot assay and ELISA were performed to identify the immunogenicity of the recombinant phage PⅢ protein-displayed and artificially-synthesized T-B com-bined antigenic epitopes, respectively.MTS assay and ELISA were performed to detect the activation of T cells and the expression of IL-2, IL-4 and IFN-γinduced by T-B combined antigenic epitopes, respectively. Results All of the tested pathogenic Leptospira strains were positive for loa22 gene, sharing 85.5%-99.8%homologies in nucleotide sequences and 93.9%-99.5%homologies in amino acid sequences.The construc-ted prokaryotic expression system for loa22 gene efficiently expressed the rLoa22 protein.Among four T-B combined antigenic epitopes (Loa22-77, Loa22-90, Loa22-125 and Loa22-157), only Loa22-90 epitope presented a strong positive band in Western blot analysis.The proliferation of CD4+T cells and the expres-sion of IL-2 ( Th1 ) and IL-4 ( Th2 ) were significantly enhanced by the stimulation with Loa22-90 epitope peptide (P<0.05).Conclusion Loa22 protein is a sequence-conserved genus-specific outer membrane protein of L.interrogans.The Loa22-90 epitope is the predominant T-B combined antigenic epitope of Loa22 protein, which might be used as a candidate antigenic epitope in the development of multiple antigenic pep-tide ( MAP) vaccines against Leptospira infection.

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