1.Development of a High-throughput Sequencing Platform for Detection of Viral Encephalitis Pathogens Based on Amplicon Sequencing
Li Ya ZHANG ; Zhe Wen SU ; Chen Rui WANG ; Yan LI ; Feng Jun ZHANG ; Hui Sheng LIU ; He Dan HU ; Xiao Chong XU ; Yu Jia YIN ; Kai Qi YIN ; Ying HE ; Fan LI ; Hong Shi FU ; Kai NIE ; Dong Guo LIANG ; Yong TAO ; Tao Song XU ; Feng Chao MA ; Yu Huan WANG
Biomedical and Environmental Sciences 2024;37(3):294-302
Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.
2.Protective effect and mechanisms of neostigmine in combination with anisodamine against pulmonary oxygen toxicity
Guangyu ZHANG ; Jing DU ; Mengzhen LIU ; Danni ZHU ; Hui YAN ; Chong LIU
Journal of Pharmaceutical Practice and Service 2024;42(10):433-438,444
Objective Pulmonary oxygen poisoning resulting from hyperbaric oxygen,frequently occurs in specialized operations,without any current effective prevention or treatment measures.To elucidate the impact and mechanism of neostigmine(NEO)in combination with anisodamine(ANI)(neoscopolamine)on pulmonary oxygen toxicity.Methods The animal model of pulmonary oxygen poisoning was established.C57BL/6 mice were exposed to 2.5 ATA 99.9%oxygen for 6 h.The control group mice were injected with normal saline ip,while the treatment group mice received injections of ANI(25 mg/kg,ip)and NEO(50 μg/kg,ip).Lung tissues were collected and stained with HE to observe any pathological injuries after exposure.Evans blue stain was utilized to identify lung permeability,wet/dry lung ratio,and protein concentration in the bronchoalveolar lavage fluid(BALF)to assess the lung injury's severity.The modifications in inflammatory factors,oxidative stress indicators,and iron content in lung tissue were assessed.Results The results showed that the 2.5 ATA 99.9%oxygen-exposed group experienced a significant worsening of lung injury,as well as increased lung permeability,lung wet/dry ratio,and protein content in alveolar lavage fluid when compared to the control group.Moreover,mRNA levels of pro-inflammatory cytokines IL-1β,IL-6,TNF-α,and IFN-γ in the lung tissue of the model group were significantly elevated,while the levels of anti-inflammatory cytokines IL-4 and TGF-β were significantly reduced.The oxidative index MDA also significantly increased,while the antioxidant index GSH significantly decreased.Additionally,the expression of GPX4,a marker of ferroptosis,increased with an increase in iron content.Neoscopolamine treatment successfully reversed those effects.Conclusion The combined use of ANI and NEO had a protective effect on pulmonary oxygen poisoning.Neoscopolamine may inhibit inflammation and oxidative stress by activating the cholinergic anti-inflammatory pathway,thereby reducing the content of free iron in lung tissue and finally inhibiting cell ferroptosis.
3.Role and mechanism of epithelial-mesenchymal transition in a rat model of bronchopulmonary dysplasia induced by hyperoxia exposure
Ya-Ting LIN ; Chong-Bin YAN ; Wen-Chao HONG ; Cheng CAI ; Xiao-Hui GONG
Chinese Journal of Contemporary Pediatrics 2024;26(7):765-773
Objective To investigate the role and mechanism of epithelial-mesenchymal transition(EMT)in a rat model of bronchopulmonary dysplasia(BPD).Methods The experiment consisted of two parts.(1)Forty-eight preterm rats were randomly divided into a normoxia group and a hyperoxia group,with 24 rats in each group.The hyperoxia group was exposed to 85%oxygen to establish a BPD model,while the normoxia group was kept in room air at normal pressure.Lung tissue samples were collected on days 1,4,7,and 14 of the experiment.(2)Rat type II alveolar epithelial cells(RLE-6TN)were randomly divided into a normoxia group(cultured in air)and a hyperoxia group(cultured in 95%oxygen),and cell samples were collected 12,24,and 48 hours after hyperoxia exposure.Hematoxylin-eosin staining was used to observe alveolarization in preterm rat lungs,and immunofluorescence was used to detect the co-localization of surfactant protein C(SPC)and α-smooth muscle actin(α-SMA)in preterm rat lung tissue and RLE-6TN cells.Quantitative real-time polymerase chain reaction and protein immunoblotting were used to detect the expression levels of EMT-related mRNA and proteins in preterm rat lung tissue and RLE-6TN cells.Results(1)Compared with the normoxia group,the hyperoxia group showed blocked alveolarization and simplified alveolar structure after 7 days of hyperoxia exposure.Co-localization of SPC and α-SMA was observed in lung tissue,with decreased SPC expression and increased α-SMA expression in the hyperoxia group at 7 and 14 days of hyperoxia exposure compared to the normoxia group.In the hyperoxia group,the mRNA and protein levels of TGF-β1,α-SMA,and N-cadherin were increased,while the mRNA and protein levels of SPC and E-cadherin were decreased at 7 and 14 days of hyperoxia exposure compared to the normoxia group(P<0.05).(2)SPC and α-SMA was observed in RLE-6TN cells,with decreased SPC expression and increased α-SMA expression in the hyperoxia group at 24 and 48 hours of hyperoxia exposure compared to the normoxia group.Compared to the normoxia group,the mRNA and protein levels of SPC and E-cadherin in the hyperoxia group were decreased,while the mRNA and protein levels of TGF-β1,α-SMA,and E-cadherin in the hyperoxia group increased at 48 hours of hyperoxia exposure(P<0.05).Conclusions EMT disrupts the tight connections between alveolar epithelial cells in a preterm rat model of BPD,leading to simplified alveolar structure and abnormal development,and is involved in the development of BPD.
4.New progress and challenges in the diagnosis and treatment of cosmetic allergy-related adverse reactions.
Chong ZHANG ; Shao Min ZHONG ; Yan WU ; Jia Hui ZHAO ; Zuo Tao ZHAO ; Yan TIAN ; Hang LI
Chinese Journal of Preventive Medicine 2023;57(4):591-596
In China, the current standard for cosmetic adverse reactions related skin disease (GB/T 17149.1-1997) was jointly issued by the Ministry of Health and the State Bureau of Technical Supervision in 1997, cosmetic-allergic adverse reactions include allergic contact dermatitis and photo-allergic contact dermatitis according to this standard. The increasing use and changes in cosmetic ingredients or formula lead to a significant increase for the incidence of adverse reactions as the cosmetics industry is developing rapidly in the last 20 years. In the meantime, the clinical manifestations have become more diverse. In recent years, there have been many reports on the special manifestations for cosmetic allergy and allergen test, which provide a reference for the subsequent improvement of the diagnosis and prevention.
Humans
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Patch Tests/adverse effects*
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Dermatitis, Allergic Contact/therapy*
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Cosmetics/adverse effects*
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China
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Incidence
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Allergens/adverse effects*
5.The distribution of blood pressure and associated factors of the elderly with type 2 diabetes in Jiangsu Province.
Jia Hui LIU ; Han Kun XIE ; Jian SU ; Zheng ZHU ; En Chun PAN ; Yan LU ; Fu Ping WAN ; Qing Yang YAN ; Ning ZHANG ; Shu Jun GU ; Ming WU ; Jin Yi ZHOU ; Chong SHEN
Chinese Journal of Preventive Medicine 2023;57(5):614-625
Objective: To investigate the distribution of blood pressure and analyze the associated factors of blood pressure of the elderly with type 2 diabetes in Jiangsu Province. Methods: The elderly over 60 years old participants with type 2 diabetes in the communities of Huai'an City and Changshu City, Jiangsu Province were selected in this study. They were divided into two groups: taking antihypertensive drugs and not taking antihypertensive drugs. The demographic characteristics, such as age and sex, and relevant factors were collected by questionnaire. The systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured by physical examination. The percentile of SBP and DBP in each age group of men and women were described. The kernel density estimation curve was used to show the blood pressure distribution. The trend of blood pressure with age was fitted by locally weighted regression. The logistic regression model was used to analyze relevant factors of blood pressure. Results: A total of 12 949 participants were included in this study, including 7 775 patients in the antihypertensive drug group and 5 174 patients in the group without antihypertensive drugs. The SBP of participants was concentrated at 140-160 mmHg, and their DBP was concentrated at 75-85 mmHg. There were significant differences in the distribution of blood pressure among the subgroups of body mass index (BMI) and rural areas whether taking antihypertensive drugs and not. For participants aged under 80 years old, the SBP showed an increasing trend with age and the DBP showed a decreasing trend with age. Age, BMI ≥24 kg/m2, fasting blood glucose ≥7.0 mmol/L, living in rural areas and no smoking were influencing factors of the elevated SBP; BMI ≥24 kg/m2, male, living in rural areas, no smoking, drinking alcohol and not receiving drug hypoglycemic treatment were influencing factors of the elevated DBP. Conclusion: The SBP of older diabetic adults in Jiangsu Province is at a high level, and the distribution of blood pressure is significantly different between men and women in taking antihypertensive drugs group. The SBP presents a rising trend and the DBP is decreasing at the age of 60-80 years. The blood pressure level of this population are mainly affected by age, BMI, urban and rural areas, smoking.
Adult
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Aged
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Humans
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Male
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Female
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Middle Aged
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Aged, 80 and over
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Blood Pressure/physiology*
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Diabetes Mellitus, Type 2/epidemiology*
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Antihypertensive Agents/therapeutic use*
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Smoking
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Body Mass Index
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Hypertension/epidemiology*
6.Dissection of Cellular Communication between Human Primary Osteoblasts and Bone Marrow Mesenchymal Stem Cells in Osteoarthritis at Single-Cell Resolution
Ying LIU ; Yan CHEN ; Xiao-Hua LI ; Chong CAO ; Hui-Xi ZHANG ; Cui ZHOU ; Yu CHEN ; Yun GONG ; Jun-Xiao YANG ; Liang CHENG ; Xiang-Ding CHEN ; Hui SHEN ; Hong-Mei XIAO ; Li-Jun TAN ; Hong-Wen DENG
International Journal of Stem Cells 2023;16(3):342-355
Background and Objectives:
Osteoblasts are derived from bone marrow mesenchymal stem cells (BMMSCs) and playimportant role in bone remodeling. While our previous studies have investigated the cell subtypes and heterogeneity in osteoblasts and BMMSCs separately, cell-to-cell communications between osteoblasts and BMMSCs in vivo in humans have not been characterized. The aim of this study was to investigate the cellular communication between human primary osteoblasts and bone marrow mesenchymal stem cells.
Methods:
and Results: To investigate the cell-to-cell communications between osteoblasts and BMMSCs and identifynew cell subtypes, we performed a systematic integration analysis with our single-cell RNA sequencing (scRNA-seq) transcriptomes data from BMMSCs and osteoblasts. We successfully identified a novel preosteoblasts subtype which highly expressed ATF3, CCL2, CXCL2 and IRF1. Biological functional annotations of the transcriptomes suggested that the novel preosteoblasts subtype may inhibit osteoblasts differentiation, maintain cells to a less differentiated status and recruit osteoclasts. Ligand-receptor interaction analysis showed strong interaction between mature osteoblasts and BMMSCs. Meanwhile, we found FZD1 was highly expressed in BMMSCs of osteogenic differentiation direction. WIF1 and SFRP4, which were highly expressed in mature osteoblasts were reported to inhibit osteogenic differentiation. We speculated that WIF1 and sFRP4 expressed in mature osteoblasts inhibited the binding of FZD1 to Wnt ligand in BMMSCs, thereby further inhibiting osteogenic differentiation of BMMSCs.
Conclusions
Our study provided a more systematic and comprehensive understanding of the heterogeneity of osteogenic cells. At the single cell level, this study provided insights into the cell-to-cell communications between BMMSCs and osteoblasts and mature osteoblasts may mediate negative feedback regulation of osteogenesis process.
7.Clinical Efficacy of Modified Sanxiaoyin in Treatment of Mild or Moderate COVID-19 Patients: Based on Retrospective Analysis
Zhi-hui LUO ; Kun-xiu WANG ; Yan-lin ZHANG ; Zi-qin CHEN ; Bei CHEN ; Jia CHEN ; Ting ZHOU ; Xiao-lei GU ; Chong-li LI ; Peng YAN ; Ling-ling TIAN ; Chi-cheng XU ; Bo-lin CHEN ; Song CHEN ; Qing QI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(1):150-156
ObjectiveTo retrospectively analyze the clinical data of 52 patients with coronavirus disease-2019 (COVID-19) and explore the clinical efficacy of modified Sanxiaoyin on mild/moderate COVID-19 patients. MethodThe propensity score matching method was used to collect the clinical data of mild or moderate COVID-19 patients enrolled in the designated hospital of the Second Hospital of Jingzhou from December 2019 to May 2020. A total of 26 eligible patients who were treated with modified Sanxiaoyin were included in the observation group, and the 26 patients treated with conventional method were the regarded as the control. The disappearance of clinical symptoms, disappearance time of main symptoms, efficacy on traditional Chinese medicine (TCM) symptoms, hospitalization duration, laboratory test indicators, and CT imaging changes in the two groups were compared. ResultThe general data in the two groups were insignificantly different and thus they were comparable. After 7 days of treatment, the disappearance rate of fever, cough, fatigue, dry throat, anorexia, poor mental state, and poor sleep quality in the observation group was higher than that in the control group (P<0.05), and the difference in the disappearance rate of expectoration and chest distress was insignificant. For the cases with the disappearance of symptoms, the main symptoms (fever, cough, fatigue, dry throat, anorexia, chest distress) disappeared earlier in the observation group than in the control group (P<0.01). After 7 days of treatment, the scores of the TCM symptom scale of both groups decreased (P<0.01), and the decrease of the observation group was larger that of the control group (P<0.01). All patients in the two groups were cured and discharged. The average hospitalization duration in the observation group [(12.79±2.68) d] was shorter than that in the control group [(15.27±3.11) d] (P<0.01). The effective rate in the observation group (92.31%, 24/26) was higher than that in the control group (76.92%, 20/26) . After 7 days of treatment, the lymphocyte (LYM) count increased (P<0.05), and white blood cell (WBC) count and neutrophil (NEUT) count decreased insignificantly in the two groups. Moreover, levels of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and procalcitonin (PCT) reduced in the two groups after treatment (P<0.01) and the reduction in the observation group was larger than that in the control group (P<0.01). Through 7 days of treatment, the total effective rate on pulmonary shadow in the observation group (90.00%, 18/20) was higher than that in the control group (77.27%, 17/22) (P>0.05) and the improvement of lung shadow in the observation group was better than that in the control group (P<0.01). ConclusionModified Sanxiaoyin can significantly alleviate fever, cough, fatigue, anorexia, chest distress, poor sleep quality, and other symptoms of patients with mild or moderate COVID-19, improve biochemical indicators, and promote the recovery of lung function. This paper provides clinical evidence for the application of modified Sanxiaoyin in the treatment of mild or moderate COVID-19.
8.Metagenomic sequencing for diagnosis of sparganosis mansoni: a case report
Chong-min PANG ; Xing-lin YANG ; Yan WANG ; Hui ZHAI ; Feng MIAO ; Shi-min ZHANG
Chinese Journal of Schistosomiasis Control 2022;34(5):556-558
The patient was found to develop a migrating mass in the lower abdomen without any known cause in 2000, and the cause had not been identified following multiple diagnoses since then. The mass was found to migrate to the left anterior axillary regions on August 11, 2020. Then, three segments of incomplete white worms were resected through minimally invasive surgery, and metagenomic sequencing revealed sparganosis mansoni. After surgical resection of complete worms was performed on October 21, 2021, the case was cured and discharged from the hospital. Follow-up revealed satisfactory outcomes and no new mass was found throughout the body.
9.Gene Mutation and Overexpression of Newly Diagnosed Multiple Myeloma Patients.
Yi FAN ; Shu-Juan WANG ; Yan-Fang LIU ; Chong WANG ; Ya-Fei LI ; Wei-Qiong WANG ; Qian-Qian HAO ; Dan-Feng ZHANG ; Ying-Mei LI ; Hui SUN ; Rong GUO ; Shao-Qian CHEN ; Xin-Sheng XIE ; Tao LI ; Ding-Ming WAN ; Zhong-Xing JIANG
Journal of Experimental Hematology 2022;30(1):166-169
OBJECTIVE:
To analyze the characteristics of gene mutation and overexpression in newly diagnosed multiple myeloma (NDMM) patients.
METHODS:
Bone marrow cells from 208 NDMM patients were collected and analyzed. The gene mutation of 28 genes and overexpression of 6 genes was detected by DNA sequencing. Chromosome structure abnormalities were detected by fluorescence in situ hybridization (FISH).
RESULTS:
Gene mutations were detected in 61 (29.33%) NDMM patients. Some mutations occurred in 5 or more cases, such as NRAS, PRDM1, FAM46C, MYC, CCND1, LTB, DIS3, KRAS, and CRBN. Overexpression of six genes (CCND1, CCND3, BCL-2, CCND2, FGFR3, and MYC) were detected in 83 (39.9%) patients, and cell cycle regulation gene was the most common. Single nucleotide polymorphisms (SNP) changes were detected in 169 (81.25%) patients, the TP53 P72R gene SNP (70.17%) was the most common. Abnormality in chromosome structure was correlated to gene overexpression. Compared to the patients with normal chromosome structure, patients with 14q32 deletion showed higher proportion of CCND1 overexpression. Similarly, patients with 13q14 deletion showed higher proportion of FGFR3 overexpression, whereas patients with 1q21 amplification showed higher proportion of CCND2, BCL-2 and FGFR3 overexpression.
CONCLUSION
There are multiple gene mutations and overexpression in NDMM. However, there is no dominated single mutation or overexpression of genes. The most common gene mutations are those in the RAS/MAPK pathway and the genes of cyclin family CCND are overexpression.
Chromosome Aberrations
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Humans
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In Situ Hybridization, Fluorescence
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Multiple Myeloma/genetics*
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Mutation
10.Clinical observation on filiform-fire needling of "Biaoben acupoint combination" for 33 cases of sequelae of coronavirus disease 2019 during recovery period.
Zhi-Hui LUO ; Kun-Xiu WANG ; Yan-Lin ZHANG ; Zi-Qin CHEN ; Bei CHEN ; Jia CHEN ; Ting ZHOU ; Xiao-Lei GU ; Chong-Li LI ; Peng YAN ; Ling-Ling TIAN ; Chi-Cheng XU ; Bo-Lin CHEN ; Song CHEN ; Feng-Xia LIANG ; Song WU
Chinese Acupuncture & Moxibustion 2022;42(7):760-764
OBJECTIVE:
To observe the clinical therapeutic effect of filiform-fire needling of "Biaoben acupoint combination" on the sequelae of patients with coronavirus disease 2019 (COVID-19) during the recovery period.
METHODS:
A total of 33 patients with COVID-19 during the recovery period were treated with filiform-fire needling at the acupoints of Mingmen (GV 4), Shenzhu (GV 12), Gaohuang (BL 43), Zusanli (ST 36) and Shangjuxu (ST 37), etc., once every other day, 3 times a week, and 3 times was one course of treatment and totally 2 courses of treatment were required. The TCM symptom, Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD) scores, pulmonary function indexes (forced vital capacity [FVC], forced expiratory volume in one second [FEV1], peak expiratory flow [PEF]) and chest CT imaging change were observed before and after treatment, and the therapeutic effect was evaluated.
RESULTS:
After treatment, the scores of TCM symptom, HAMA and HAMD were decreased compared with those before treatment (P<0.05), and the levels of FVC, FEV1 and PEF were increased compared with those before treatment (P<0.05), and the recovery rate of 22 patients with pulmonary ventilation dysfunction was 86.4% (19/22). After treatment, the lung shadow area was smaller than that before treatment (P<0.05). The effective rate of 25 patients with lung CT abnormalities was 84.0% (21/25). After treatment, 23 cases were cured, 5 cases were markedly effective, 4 cases were effective, 1 case was ineffective, the cured and markedly effective rate was 84.8%.
CONCLUSION
The filiform-fire needling of "Biaoben acupoint combination" could significantly reduce the sequelae of cough, fatigue, chest tightness, etc. and mental symptoms such as anxiety and depression in patients with COVID-19 during the recovery period, and promote inflammatory exudation absorption of pulmonary lesion and improve lung ventilation function.
Acupuncture Points
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Acupuncture Therapy
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COVID-19/therapy*
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Humans
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Lung
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Vascular Surgical Procedures

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