Online First

Articles in press have been peer-reviewed and accepted, which are not yet assigned to volumes/issues, but are citable by Digital Object Identifier (DOI).
Original Article
An Investigation of the Effects of B7-H4 Gene rs10754339 and miR-125a Gene rs12976445 on Cancer Susceptibility
JIN Yu Chen, DONG Li Juan, YANG Qin Yue, XIONG Wei Ning, WANG Wei Yi, FENG Xian Hong, YU Wei, HUANG Wei, CHEN Bi Feng
, Available online  , doi: 10.3967/bes2023.074
  Objective  To investigate the effects of the B7-H4 gene rs10754339 and miR‐125a gene rs12976445 on cancer susceptibility through a case-control study and meta-analysis.   Methods  A total of 1,490 cancer patients (lung/gastric/liver/: 550/460/480) and 800 controls were recruited in this case-control study. The meta-analysis was performed by pooling the data from previous related studies and the present study.   Results  The results of this study showed that in the Hubei Han Chinese population, the rs10754339 gene was significantly associated with the risk of lung and gastric cancer but not liver cancer, and the rs12976445 gene was significantly associated with the risk of lung cancer but not liver or gastric cancer. The meta-analysis results indicated that rs10754339 and rs12976445 contributed to cancer susceptibility in the Chinese population and also revealed a significant association between rs10754339 and breast cancer risk, as well as between rs12976445 and lung cancer risk.   Conclusion  The B7-H4 gene rs10754339 and miR‐125a gene rs12976445 may be the potential genetic markers for cancer susceptibility in the Chinese population, which should be validated in future studies with larger sample sizes in other ethnic populations.
Treatment Outcomes in COVID-19 Patients with Brucellosis: Case Series in Heilongjiang and Systematic Review of Literature
YANG Man Li, WANG Jing Ya, ZONG Xing Yu, GUAN Li, LI Hui Zhen, Xiong Yi Bai, LIU Yu Qin, LI Ting, JI Xin Yu, SHANG Xi Yu, ZHANG Hui Fang, GUO Yang, GONG Zhao Yuan, ZHANG Lei, TONG Lin, Chen Ren Bo, FAN Yi Pin, QIN Jin, WANG Fang, LIN Gang, SHI Nan Nan, WANG Yan Ping, MA Yan
, Available online  , doi: 10.3967/bes2023.035
  Objective  Clinical characteristics and outcome in COVID-19 with brucellosis patients has not been well demonstrated, we tried to analyze clinical outcome in local and literature COVID-19 cases with brucellosis before and after recovery.   Methods  We retrospectively collected hospitalization data of comorbid patients and prospectively followed up after discharge in Heilongjiang Infectious Disease Hospital from January 15, 2020 to April 29, 2022. Demographics, epidemiological, clinical symptoms, radiological and laboratory data, treatment medicines and outcomes, and follow up were analyzed, and findings of a systematic review were demonstrated.   Results  A total of four COVID-19 with brucellosis patients were included. One patient had active brucellosis before covid and 3 patients had nonactive brucellosis before brucellosis. The median age was 54.5 years, and all were males (100.0%). Two cases (50.0%) were moderate, and one was mild and asymptomatic, respectively. 3 cases (75.0%) had at least one comorbidity (brucellosis excluded). All 4 patients were found in COVID-19 nucleic acid screening. Case C and D had only headache and fever on admission, respectively. Four cases were treated with Traditional Chinese medicine, western medicines for three cases, no adverse reaction occurred during hospitalization. All patients were cured and discharged. Moreover, one case (25.0%) had still active brucellosis without re-positive COVID-19, and other three cases (75.0%) have no symptoms of discomfort except one case fell fatigue and anxious during the follow-up period after recovery. Conducting the literature review, two similar cases have been reported in two case reports, and were both recovered, whereas, no data of follow up after recovery.   Conclusions  These cases indicate that COVID-19 patients with brucellosis had favorable outcome before and after recovery. More clinical studies should be conducted to confirm our findings.
Characterization of circRNA-associated ceRNA networks in folate deficiency-induced neural tube defects
WANG Shan, ZENG Yu Bing, PEI Pei, HE Xue Jia, LIU Fan, WANG Yi, ZHANG Ting
, Available online  , doi: 10.3967/bes2023.071
  Objective  Circular RNAs (circRNAs) participate in several important pathological processes and have been used in the diagnosis and treatment of various diseases. This study aimed to investigate the role of circRNAs in neural tube defects (NTDs).   Method  we characterized circRNA-associated competitive endogenous RNA (ceRNA) networks in brain tissue of low folate -induced NTDs mouse at embryonic day 13.5 by high‐throughput sequencing. The expression levels of Circzfp644, miR-20-5p and Gas7 were detected by RT-PCR. Gas7 and Circzfp644 functions were determined by miRNA-mimics and inhibitors in mouse teratocarcinoma cells (F9 cells), and luciferase gene reporter assay was assessed in the F9 cells. In addition, the expression levels of Circzfp644, miR-20-5p and Gas7 were determined by Nanostring in human NTDs tissues.   Results  We detected 57 circRNA transcripts, 16 miRNAs, and 148 mRNAs that were significantly dysregulated in NTDs brain tissues compared with their expression levels in control (normal) tissues. Circzfp644 shared miRNA response elements with the growth arrest specific 7 (Gas7) gene and competitively bound with miR-20-5p to increase the expression of Gas7. Downregulation of Circzfp644 and Gas7 and upregulation of miR-20-5p were found in human NTD tissue.   Conclusion  This study provides new perspectives on the role of circRNAs in nervous system development and the pathogenesis of NTDs.
Establishment and Modification of Ninety-seven Pneumococcal Serotyping Assays Based on Quantitative Real-time Polymerase Chain Reaction
CHE Jie, CHEN Bo Han, XU Li, GAO Yuan, YUE Meng Meng, CHEN Zi Man, ZHANG Mao Jun, SHAO Zhu Jun
, Available online  , doi: 10.3967/bes2023.078
  Objective  To establish and modify quantitative real-time polymerase chain reaction (qPCR)-based serotyping assays to distinguish 97 pneumococcal serotypes. Methods A database of capsular polysaccharide (cps) loci sequences was generated, covering 97 pneumococcal serotypes. Bioinformatics analyses were performed to identify the cps loci structure and target genes related to different pneumococcal serotypes with specific SNPs. A total of 27 novel qPCR serotyping assay primers and probes were established based on qPCR, while 27 recombinant plasmids containing serotype-specific DNA sequence fragments were constructed as reference target sequences to examine the specificity and sensitivity of the qPCR assay. A panel of pneumococcal reference strains was employed to evaluate the capability of pneumococcal serotyping.   Results  A total of 97 pneumococcal serotyping assays based on qPCR were established and modified, which included 64 serotypes previously reported as well as an additional 33 serotypes. Twenty-seven novel qPCR serotyping target sequences were implemented in the pneumococcal qPCR serotyping system. A total of 97 pneumococcal serotypes, which included 52 individual serotypes and 45 serotypes belonging to 20 serogroups, could not be identified as individual serotypes. The sensitivity of qPCR assays based on 27 target sequences was 1–100 copies/µL. The specificity of the qPCR assays was 100%, which were tested by a panel of 90 serotypes of the pneumococcal reference strains.   Conclusions  A total of 27 novel qPCR assays were established and modified to analyze 97 pneumococcal serotypes.
Protein Containing the GGDEF Domain Affects Motility and Biofilm Formation in Vibrio cholerae and is Negatively Regulated by Fur and HapR
GAO He, MA Li Zhi, QIN Qin, CUI Yao, MA Xiao Han, ZHANG Yi Quan, KAN Biao
, Available online  , doi: 10.3967/bes2023.080
  Objective  This study aimed to investigate whether the VCA0560 gene acts as an active diguanylate cyclase (DGC) in Vibrio cholerae and how its transcription is regulated by Fur and HapR.   Methods  The roles of VCA0560 was investigated by utilizing various phenotypic assays, including colony morphological characterization, crystal violet staining, Cyclic di-GMP (c-di-GMP) quantification, and swimming motility assay. The regulation of the VCA0560 gene by Fur and HapR was analyzed by luminescence assay, electrophoretic mobility shift assay, and DNase I footprinting.   Results  VCA0560 gene mutation did not affect biofilm formation, motility, and c-di-GMP synthesis in V. cholerae, and its overexpression remarkably enhanced biofilm formation and intracellular c-di-GMP level but reduced motility capacity. The transcription of the VCA0560 gene was directly repressed by Fur and the master quorum sensing regulator HapR.   Conclusion  Overexpressed VCA0560 functions as an active DGC in V. cholerae, and its transcription is repressed by Fur and HapR.
HIV-1 Subtype Diversity and Factors Affecting Drug Resistance among Patients with Virologic Failure in Antiretroviral Therapy in Hainan Province, China, 2014–2020
YU De E, XU Yu Jun, LI Mu, YANG Yuan, LIANG Hua Yue, ZHONG Shan Mei, QIN Cai, LAN Ya Nan, LI Da Wei, YU Ji Peng, PANG Yuan, QIN Xue Qiu, LIANG Hao, ZHU Kao Kao, YE Li, LIANG Bing Yu
, Available online  , doi: 10.3967/bes2023.077
  Objective  This study aimed to determine the HIV-1 subtype distribution and HIV drug resistance (HIVDR) in patients with ART failure from 2014 to 2020 in Hainan, China.   Methods  A 7-year cross-sectional study was conducted among HIV/AIDS patients with ART failure in Hainan. We used online subtyping tools and the maximum likelihood phylogenetic tree to confirm the HIV subtypes with pol sequences. Drug resistance mutations (DRMs) were analyzed using the Stanford University HIV Drug Resistance Database.   Results  A total of 307 HIV-infected patients with ART failure were included, and 241 available pol sequences were obtained. Among 241 patients, CRF01_AE accounted for 68.88%, followed by CRF07_BC (17%) and eight other subtypes (14.12%). The overall prevalence of HIVDR was 61.41%, and the HIVDR against NNRTIs, NRTIs, and PIs were 59.75%, 45.64%, and 2.49%, respectively. Unemployed patients, hypoimmunity, opportunistic infections in individuals, and samples from 2017 to 2020 increased the odd ratios of HIVDR. Also, HIVDR was less likely to affect female patients. The common DRMs to NNRTIs were K103N (21.99%) and Y181C (20.33%), and M184V (28.21%) and K65R (19.09%) were the main DRMs against NRTIs.   Conclusion  The present study highlights the HIV-1 subtype diversity in Hainan and the importance of HIVDR surveillance over a long period.
Comparison of Thyroglobulin and Thyroid Function in Pregnant Women between Counties with a Median Urinary Iodine Concentration of 100–149 µg/L and 150–249 µg/L
CHEN Di Qun, YE Ying, WU Jia Ni, LAN Ying, WANG Mu Hua, WU Xiao Yan, HE Meng, WANG Li Jin, Zheng Xin Yi, CHEN Zhi Hui
, Available online  , doi: 10.3967/bes2023.076
  Objective   This study explored whether thyroglobulin and thyroid disease prevalence rates were higher in pregnant Chinese women with a median urinary iodine concentration of 100–149 µg/L , compared with those with a median urinary iodine concentration of 150–249 μg/L maintained through sustainable universal salt iodization.  Methods   This was a cross-sectional study in which 812 healthy pregnant women were enrolled to collect samples of their household edible salt, urine, and blood during their routine antenatal care in the 18 counties in Fujian Province, China. The levels of salt iodine concentration, urinary iodine concentration (UIC), free triiodothyronine (FT3), free thyroid hormone (FT4), thyroid-stimulating hormone (TSH), thyroglobulin (Tg), thyroid peroxidase antibody and thyroglobulin antibody were assessed during the routine antenatal care visits.  Results   The median UIC (mUIC) in pregnant women was 130.8 μg/L (interquartile range = 91.5–198.1 μg/L) in the counties with an mUIC of 100–149 μg/L (Group I), and 172.0 μg/L (interquartile range = 123.5–244.4 μg/L) in the counties with an mUIC of 150–249 μg/L (GroupⅡ). Goiter prevalence and thyroid nodule detection rates showed no difference between Group I and GroupⅡ(P > 0.05). Except for FT4 values, the TSH, FT4, FT3, Tg and Tg values > 40 (μg/L) and the thyroid diseases prevalence rate (TDR) showed no significant differences between Group I and GroupⅡ (P > 0.05), whether or not iodine supplementation measures were taken.  Conclusions   Compared with an mUIC of 150–249 μg/L, not only there was no difference in thyroid morphology, but also the Tg value, rate of Tg values > 40 µg/L, and TDR were not higher in pregnant women in the counties with an mUIC of 100–149 μg/L achieved through sustainable universal salt iodization in Fujian Province, China.
A Comprehensive View on the Progress of Organoid Research with an Emphasis on its Relevance to Disease Characterization
Chandra Kishore, Vaishali Ji, Saurav Mallik, Ayan Mukherji, Namrata Tomar, Soumen Kumar Pati, Aimin Li, Sinthia Roy Banerjee, Soumadip Ghosh, Raza Ali Naqvi
, Available online  , doi: 10.3967/bes2023.083
Lung cancer risk attributable to active smoking in China: a systematic review and meta-analysis
ZHAO Jian, SHI Yu Lin, WANG Yu Tong, AI Fei Ling, WANG Xue Wei, YANG Wen Yi, WANG Jing Xin, AI Li Mei, HU Kui Ru, WAN Xia
, Available online  , doi: 10.3967/bes2023.075
  Objective  No consensus exists on the relative risk (RR) of lung cancer (LC) attributable to active smoking in China. This study aimed to evaluate the unified RR of LC attributable to active smoking among the Chinese population.   Methods  A systematic literature search of seven databases was conducted to identify studies reporting active smoking among smokers versus nonsmokers in China. Primary articles on LC providing risk estimates with their 95% confidence intervals (CIs) for “ever,” “former,” or “current” smokers from China were selected. Meta-analysis was used to estimate the pooled RR of active smoking.   Results  Forty-four unique studies were included. Compared with that of nonsmokers, the pooled RR (95% CI) for “ever,” “former,” and “current” smokers were 3.26 (2.79–3.82), 2.95 (1.71–5.08), and 5.16 (2.58–10.34) among men, 3.18 (2.78–3.63), 2.70 (2.08–3.51), and 4.27 (3.61–5.06) among women, and 2.71 (2.12–3.46), 2.66 (2.45–2.88), and 4.21 (3.25–5.45) in both sexes combined, respectively.   Conclusion  The RR of LC has remained relatively stable (range, 2–6) over the past four decades in China. Early quitting of smoking could reduce the RR to some extent; however, completely refraining from smoking is the best way to avoid its adverse effects.