Meta-analysis of InterTan, PFNA and PFNA-II internal fixation for the treatment of unstable intertrochanteric fractures in elderly individuals.

Elderly individuals are often affected by osteoporosis and have poor stability after fracture reduction. Moreover, there is still controversy regarding the clinical effects of the treatment for unstable intertrochanteric fractures in the elderly. The Cochrane, Embase, PubMed, and other databases were searched, and a meta-analysis of the literature on the treatment of unstable intertrochanteric fractures of the elderly with InterTan, PFNA, and PFNA-II was conducted. Seven studies were screened, with a total of 1236 patients. Our meta-analysis results show that InterTan is not significantly different from PFNA in terms of operation and fluoroscopy times, but it takes longer than PFNA-II. In terms of postoperative screw cut, pain, femoral shaft fracture, and secondary operations, InterTan is superior to PFNA and PFNA-II. Conversely, in terms of intraoperative blood loss, hospital stay, and postoperative Harris score, there is no significant difference between InterTan and PFNA and PFNA-II. Compared to PFNA and PFNA-II, InterTan internal fixation has advantages in the treatment of unstable intertrochanteric fractures in elderly individuals in terms of screw cutting, femoral shaft fractures, and secondary operations. However, InterTan operation and fluoroscopy times take longer than PFNA and PFNA-II.

[Effect of autologous hematopoietic stem cell transplantation on minimal residual disease in patients with multiple myeloma].

Objective: To evaluate the effect of autologous hematopoietic stem cell transplantation (ASCT) on minimal residual disease (MRD) in patients with multiple myeloma (MM). Method: From August 2018 to August 2021, 92 patients newly diagnosed with MM who had received either the bortezomib combined with cyclophosphamide and dexamethasone (VCD) or the bortezomib, lenalidomide and dexamethasone (VRD) induction regimens followed by sequential ASCT were assessed for overall survival (OS) and the MRD negative rate. The differences in efficacy at 100 days after transplantation were assessed according to factors, including age, risk stratification, target organ damage, and pre-transplant regimen, etc. Results: Among the 92 patients, there were 45 males and 47 females, with a median age of 57.3 (35-67) years. Fifty-seven patients received the VCD regimen, and 35 received VRD as induction regimen. Forty-three patients received busulphan combined with cyclophosphamide and etoposide (BCV), and 49 patients received high-dose melphan (HDM) regimen as pre-transplantation treatment. After transplantation, the total complete remission (CR) rate of 92 patients increased from 23.9% (22/92) to 58.7% (54/92), and the MRD negative rate increased from 4.4% (4/92) to 33.7% (31/92), and the differences were statistically significant (all P<0.05). After transplantation, the MRD negative rates of patients with PR, VGPR and ≥CR before transplantation were 17.6% (6/34), 33.3% (12/36) and 59.1% (13/22), respectively (P=0.006). The CR rates of patients with or without plasmacytoma at initial diagnosis were 36.4% (4/11) and 65.4% (53/81), respectively (P=0.029), and the MRD negative rates were 18.2% (2/11) and 39.5% (32/81), respectively (P=0.037), and the differences were statistically significant. The MRD negative rates in high-risk patients and standard-risk group were 30.5% (12/28) and 42.9% (18/59), respectively (P=0.258). For patients who achieved efficacy above VGPR before transplantation, the MRD negative rates after transplantation in VCD-induced group and VRD group were 29% (9/31) and 59.3% (16/27), respectively (P=0.033), and in BCV group and HDM group were 24% (6/25) and 57.6% (19/33), respectively (P=0.016), the differences between the groups were both statistically significant. Conclusion: ASCT can overcome the adverse factors such as high-risk cytogenetic abnormalities, and significantly improve the CR rate and MRD negative rate of MM patients. However, the benefit for patients with plasmacytoma at initial diagnosis is not as good as that of patients without.

RNA-seq of circular RNAs identified circPTPN22 as a potential new activity indicator in systemic lupus erythematosus.

BACKGROUND: Circular RNAs (circRNAs) are possible biomarkers for many diseases, but the knowledge of circRNAs in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE) remains limited. This study aimed to assess the expression of circRNAs in PBMCs from patients with SLE and healthy individuals by RNA sequencing (RNA-seq). METHODS: In total, 128 circRNAs were significantly differentially expressed including 39 upregulated and 89 downregulated circRNAs in four new-onset SLE patients compared with three healthy controls. After verification of the four candidate circRNAs in 49 patients with SLE and 37 controls using quantitative real-time polymerase chain reaction (qRT-PCR) assays, a previously undescribed circRNA with potential translation activity, circPTPN22, was selected to confirm its clinical significance. RESULTS: Bioinformatics analysis demonstrated that the parent gene of circPTPN22 was protein tyrosine phosphatase non-receptor type 22 (PTPN22), a potent regulator of T cell activation. The downregulation of circPTPN22 in patients with SLE was strongly negatively correlated with their Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) scores. circRNA-miRNA-mRNA co-expression network analysis indicated a correlation between circPTPN22 and the miRNAs and mRNAs related to immunological regulation including the development of SLE. Patients with higher SLEDAI scores had lower circPTPN22 expression levels, and long-term hormone treatment significantly increased circPTPN22 levels. Receiver operating characteristic curve analysis indicated that circPTPN22 has good diagnostic value for SLE. CONCLUSION: Our data demonstrated the aberrant expression of circRNAs in patients with SLE compared with healthy controls; circPTPN22 might function as a diagnostic and disease severity indicator in SLE.

Rapid local acceleration of relativistic radiation-belt electrons by magnetospheric chorus.

Recent analysis of satellite data obtained during the 9 October 2012 geomagnetic storm identified the development of peaks in electron phase space density, which are compelling evidence for local electron acceleration in the heart of the outer radiation belt, but are inconsistent with acceleration by inward radial diffusive transport. However, the precise physical mechanism responsible for the acceleration on 9 October was not identified. Previous modelling has indicated that a magnetospheric electromagnetic emission known as chorus could be a potential candidate for local electron acceleration, but a definitive resolution of the importance of chorus for radiation-belt acceleration was not possible because of limitations in the energy range and resolution of previous electron observations and the lack of a dynamic global wave model. Here we report high-resolution electron observations obtained during the 9 October storm and demonstrate, using a two-dimensional simulation performed with a recently developed time-varying data-driven model, that chorus scattering explains the temporal evolution of both the energy and angular distribution of the observed relativistic electron flux increase. Our detailed modelling demonstrates the remarkable efficiency of wave acceleration in the Earth's outer radiation belt, and the results presented have potential application to Jupiter, Saturn and other magnetized astrophysical objects.

[SIVA1 Regulates the Stability of Single-Stranded DNA-Binding Protein 3 Isoforms].

The assembly of LIM-homeodomain (LIM-HD) transcriptional complex plays important roles in early neuronal development. The stability of LIM-HD is controlled by single-strand binding protein 3 (SSBP3) via a cascade mechanism protecting it from proteasomal degradation. The expression level of SSBP3 has to be precisely regulated. Although a decrease of SSBP3 level is associated with several diseases, the mechanism of SSBP3 downregulation and whether SSBP3 itself is subject to proteasomal degradation remain largely unknown. Two strongly conserved transcripts of the SSBP3 gene, SSBP3a and SSBP3c, were cloned from a human brain cDNA library. By RT-PCR, we show that Ssbp3c is continuously expressed in both embryonic and adult mouse brain, whereas Ssbp3a is restricted to embryonic brain tissue. By co-IP and GST pulldown assays, we identified SIVA1 as a novel SSBP3-binding factor. In a ubiquitination assay, we show that SIVA1 enhances the ubiquitination of SSBP3 and regulates its abundance. Our findings reveal the proteasomal degradation of SSBP3 for the first time and provide a rationale for an SIVAl-SSBP3-dependent mechanism for the disassembly of LIM-HD multiprotein complexes.

Aetiological characteristics of adult acute diarrhoea in a general hospital of Shanghai.

Epidemic surveillance is an effective means to determine the characteristics of acute diarrhoea and the benefits of disease control and prevention. The epidemiological, clinical, and aetiological data of adult (aged ⩾15 years) acute diarrhoea in a general hospital in Shanghai were collected and analysed. Out of 2430 acute diarrhoea patients, 162 subjects were sampled (sample ratio 15:1). The sampled subjects had an average age (±s.d.) of 44 ± 18 years; 142 (87·7%) had a history that indicated ingestion of contaminated food; and 40 (24·7%), 54 (33·3%), and 73 (45·1%) patients had diarrhoea that was attributed to viral, bacterial, and unknown aetiological origins respectively. Viral diarrhoea is mainly prevalent during the winter and spring months, while bacterial and diarrhoea of unknown aetiology occur mainly in the summer months. The average age of the unknown aetiology group (48 ± 19 years) was significantly older than that of the viral diarrhoea group (39 ± 16 years). The number of patients with vomiting in the viral group (30·6%) was significantly higher than that in the bacterial (17·1%) and unknown aetiology (8·2%) groups. Viral and bacterial infections are the main cause of acute diarrhoea in Shanghai. However, further effective technological means are needed to improve the surveillance, control, and prevention of acute diarrhoea.

The increased expression of TCF3 is correlated with poor prognosis in Chinese patients with nasopharyngeal carcinoma.

OBJECTIVES: Regulatory factors controlling stem cell identity and self-renewal are often active in aggressive cancers and are thought to promote cancer growth and progression. B-cell-specific transcription factor 3 (TCF3/E2A) is a member of the T-cell factor/lymphoid enhancer factor (TCF/LEF) transcription factor family that is central to regulating epidermal and embryonic stem cell identity. It has been reported that TCF3 was connected with the development and progression of a number of human cancers. In this study, we aimed to identify the expression of TCF3 in human nasopharyngeal carcinoma (NPC) and evaluate its clinical significance. DESIGN: To investigate the expression of TCF3 in NPC and its relationship to prognosis. SETTING: An in vitro study. MAIN OUTCOME MEASURES: We analysed the expression of TCF3 in NPC and in non-tumourous nasopharyngeal tissues by quantitative RT-PCR and Western blotting. The expression patterns of TCF3 in 117 archived paraffin-embedded NPC specimens were characterised by immunohistochemistry, and the correlation between the TCF3 protein expression and the clinicopathological features of NPC was analysed. RESULTS: We observed that TCF3 had a higher expression in NPC than in non-tumourous nasopharyngeal tissues of 117 archived paraffin-embedded NPC specimens, and 80 (68.4%) biopsy tissues revealed high levels of TCF3 expression. Furthermore, statistical analyses demonstrated that the increased expression of TCF3 was closely related to clinical stage, locoregional recurrence and distant metastasis of NPC. NPC patients with high levels of TCF3 expression had a shorter survival time, whereas patients with lower levels of TCF3 expression survived longer. Moreover, multivariate analysis suggested that the upregulation of TCF3 was a critical prognostic factor for NPC. CONCLUSIONS: Our observations suggest, for the first time, that TCF3 is significantly associated with the development and progression of NPC, which can be used as an important prognostic marker for patients with NPC and may be an effective target for the treatment of NPC.

Apoptosis induced by lipid-associated membrane proteins from Mycoplasma hyopneumoniae in a porcine lung epithelial cell line with the involvement of caspase 3 and the MAPK pathway.

Lipid-associated membrane proteins (LAMPs) are important in the pathogenicity of the Mycoplasma genus of bacteria. We investigated whether Mycoplasma hyopneumoniae LAMPs have pathogenic potential by inducing apoptosis in a St. Jude porcine lung epithelial cell line (SJPL). LAMPs from a pathogenic strain of M. hyopneumoniae (strain 232) were used in the research. Our investigation made use of diamidino-phenylindole (DAPI) and acridine orange/ethidium bromide (AO/EB) staining, terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) analysis, and Annexin-V-propidium iodide staining. After LAMP treatment for 24 h, typical changes were induced, chromosomes were concentrated, apoptotic bodies were observed, the 3'-OH groups of cleaved genomes were exposed, and the percentage of apoptotic cells reached 36.5 ± 11.66%. Caspase 3 and caspase 8 were activated and cytochrome c (cyt c) was released from the mitochondria into the cytoplasm; poly ADP ribose polymerase (PARP) was digested into two fragments; p38 mitogen-activated protein kinase (MAPK) was phosphorylated; and the expression of pro-apoptosis protein Bax increased while the anti-apoptosis protein Bcl-2 decreased. LAMPs also stimulated SJPL cells to produce nitric oxide (NO) and superoxide. This study demonstrated that LAMPs from M. hyopneumoniae can induce apoptosis in SJPL cells through the activation of caspase 3, caspase 8, cyt c, Bax, and p38 MAPK, thereby contributing to our understanding of the pathogenesis of M. hyopneumoniae, which should improve the treatment of M. hyopneumoniae infections.

Genetic affinities of central China populations.

Hunan locates in the south-central part of China, to the south of the middle reaches of the Yangtze River and south of Lake Dongting. According to the historical records, the peopling of Hunan by modern human ancestors can ascend to 40 thousand years ago. Thus, to trace the ancient maternal components can offer further insight into the origin of south-central China. In this study, we investigated the mitochondrial DNA of 114 individuals from Hunan Province (including 34 Han, 40 Tujia and 40 Miao). Hypervariable regions I and II of the mtDNA control region were sequenced, and the relative diagnostic variations in coding region according to the updated worldwide phylogeny tree were selected and typed by restriction fragment length polymorphism analysis or direct sequencing. All individuals were classified into specific (sub)haplogroups. By comparison with the surrounding populations, southern China-prevalent haplogroups were detected with relative higher frequency in the Tujia and Miao ethnic populations, such as haplogroup B, with more than 20%, lacking in the Han population, which illustrated its southern origin characters. In addition, we also detected northern of East Asia prevalent haplogroups with a relative higher frequency in Tujia populations than in the Miao and Yao ethnic groups, implying a gene flow from Han populations. However, the language-clustering tendency was supported by our principal component analysis and further genetic estimation results. Han and ethnic groups in central China exhibited specific ancestors related to their closer language affinity, although there was extensively genetic admixture between Han and ethnic groups.

Reduced expression of insulin-like growth factor 1 receptor leads to accelerated intervertebral disc degeneration in mice.

Insulin-like growth factor 1 (IGF-1) and its receptor (insulin-like growth factor 1 receptor, IGF1R) can regulate the extracellular matrix synthesis and play a crucial role in maintaining the normal functions of the intervertebral disc (IVD). The objective of this study was to investigate whether there would be accelerated IVD degeneration (IVDD) in IGF1R+/- mice. Three IGF1R+/- male mice and three wild-type male mice were sacrificed respectively at 6, 12, and 18 weeks after birth. Six lumbar disc samples were harvested from each mouse, with a total of 54 disc samples taken from each genotype. Histomorphological analysis for the IVD was performed to assess the degenerative extent according to the classification system proposed by Boos et al. Quantitative real-time PCR and semi-quantitative histologic scoring (HScore) for immunohistochemical staining were used to evaluate the expression level of type-II collagen, aggrecan and matrix metallopeptidase 13 (MMP-13). Histomorphological analysis for the discs revealed significantly less amounts of proteoglycan and type-II collagen, and significantly higher total degenerative score in IGF1R+/- mice than in wild-type mice. Real-time PCR showed that the mRNA expressions of type-II collagen and aggrecan in the discs were significantly lower, while MMP-13 was significantly higher in IGF1R+/- mice than in wild-type mice. The results of HScore analysis were similar to those obtained from the quantitative real-time PCR. Taken together, our study indicates that reduced expression of IGF1R would lead to accelerated degeneration of IVD. IGF1R+/- mice could be regarded as a good animal model to study IVD degeneration (IVDD), and studies on the IVD of IGF1R+/- mice could provide further insight into the pathogenesis of IVDD.

Microbial screening of unrelated cord blood units in a Chinese cord blood bank.

BACKGROUND: Given limited sample volume available for sterility testing, optimal testing methods and algorithms of cord blood (CB) have not been established according to standards. STUDY DESIGN AND METHODS: The volume of CB unit was reduced to 20 mL in a closed system. Hetastarch and cryopreservation solution was added to the processed CB unit in Class 100 environment. In the routine sterility testing, 20 mL of red blood cells (RBC) sample was cultured in aerobic and anaerobic culture bottles. In verification post-thaw culture of the final product, some of the discarded final product units were also sampled and cultured after thaw. All the culture bottles were incubated in the BacT/ALERT 3D system. RESULTS: In the routine sterility testing, 139 of 7032 CB units (1·98%) were contaminated with microorganism. In 84 of these 139 units (60·4%) only the anaerobic bottle was positive. Lactobacillus spp. were the most prevalent contaminant. Sixty-two discarded CB stem cell units were recultured after thaw. Of these, 10 of 48 units with a positive culturing of RBC sample were negative in the post-thaw reculture. One of 14 units with negative culturing of RBC sample was contaminated with Bifidobacterium breve in special verification test. CONCLUSION: Our study demonstrates the predominant organisms implicated in CB microbial contamination were part of the human intestinal and vaginal flora. The larger sample volume and anaerobic culture would significantly increase the detection rate of microbial contaminated CB. We also found that potential transfusion-transmitted bacterial infection risk still existed in final product although microbial screening was performed.

Physio-pathological effects of m6A modification and its potential contribution to melanoma.

Methylation of N6-adenosine (m6A) is the most prevalent internal RNA modification and is especially common among the messenger RNAs. These m6A modifications regulate splicing, translocation, stability and translation of RNA through dynamic and reversible interactions with m6A-binding proteins, namely the writers, erasers and readers. RNA methyltransferases catalyze the m6A modifications, while demethylases reverse this methylation. Deregulation of the m6A modification process has been implicated in human carcinogenesis, including melanoma-which carries one of the highest mutant rates. In this review, we provide an up-to-date summary of m6A regulation and its biological impacts on normal and cancer cells, with emphasis on the deregulation of m6A modification and m6A regulators in melanoma. In addition, we highlight the prospective potential of exploiting m6A modification in the treatment of melanoma and non-cancer diseases.

[Identification and inner relation between gastrointestinal stromal tumor and intra-abdominal desmoid tumor].

Intra-abdominal desmoid tumor (IADT) and gastrointestinal stromal tumor (GIST) are both mesenchymal tumors mostly found in gastrointestinal tracts and easily misdiagnosed, which would directly damage the survival prognosis and quality of life of patients. With the advent of the era of precision medicine, the understanding of the above two diseases is more in-depth, and the requirements for accurate diagnosis and individualized precision treatment are more stringent. Moreover, there seems to be some internal relationship between IADT and GIST, and the lack of systematic research and discussion makes clinical decision-making and patient management easy to fall into traps and misunderstandings. Therefore, this paper reviews the clinical characteristics, pathogenesis and treatments of the two, and explore their differences and internal relations, so as to provide research and practical reference for promoting more precise and individualized diagnosis and treatment regimens.

ILC3 cells promote the proliferation and invasion of pancreatic cancer cells through IL-22/AKT signaling.

PURPOSE: Type 3 innate lymphocytes (ILC3s) are reported to be involved in lung cancer, possibly by producing interleukin-22 (IL-22). However, whether ILC3s and their secreted IL-22 molecules contribute to the pathogenesis of pancreatic cancer (PC) remains unclear. To this end, in this study, we investigated the effects and possible mechanisms of ILC3s on PC pathogenesis. METHOD: The IL-22 and IL-2i2R levels and the ILC3s' frequency in cancer tissues from PC patients and in peripheral blood from PC patients and healthy controls were analyzed by flow cytometry, immunochemistry, or immunofluorescence. The effects of IL-22-induced AKT signaling on the proliferation, invasion, and migration of PC cells were examined by co-culturing PC cell lines with ILC3s isolated from PC tissues, with or without the addition of neutralizing IL-22 antibody, IL-22R antibody or AKT inhibitor. RESULTS: Our results showed that IL-22 and ILC3s were significantly upregulated in the PBMCs and cancer tissues of PC patients, and the IL-22R level was increased in PC cells. The increased frequency of ILC3s was positively correlated with the clinical features of PC patients. Co-culture experiments indicated that ILC3s promoted the proliferation, invasion, and migration of PC cell lines by secreting IL-22 to activate AKT signaling because IL-22/IL-22R or AKT blockage markedly counteracted such effects on PC cells. CONCLUSION: Our data demonstrated that ILC3s may promote PC pathogenesis through IL-22/IL-22R-AKT signaling, suggesting a potential intervention target for PC treatment in the future.

Identification of PTK7 as a promising therapeutic target for thyroid cancer.

OBJECTIVE: To evaluate the possible involvement of PTK7 in the progression of human thyroid cancer and assess its potential effects on the proliferation and apoptosis of thyroid cancer. PATIENTS AND METHODS: Immunohistochemical (IHC) assays and clinical significance analysis were performed to explore the correlations between PTK7 expression and clinical characteristics of patients with thyroid cancer. Quantitative PCR assays and Immunoblot assays were performed to detect the expression of PTK7 in control or PTK7 shRNA plasmids transfected thyroid cancer cells. MTT assays were performed to detect the effects on the proliferation of thyroid cancer cells. Flow cytometry (FCM) assays were performed to assess the changes in cell apoptosis of thyroid cancer. Additionally, the effects of PTK7 on tumor growth were detected through in vivo tumor growth assays. RESULTS: PTK7 is highly expressed in human thyroid cancer tissues, and its expression levels are associated with the clinical characteristics, including TNM stage (p=0.015*), and intraglandular dissemination (p=0.024*) of patients with thyroid cancer. PTK7 ablation inhibits cell proliferation and stimulates cell apoptosis of thyroid cancer in vitro. Additionally, PTK7 contributes to tumor growth of thyroid cancer cells in mice. CONCLUSIONS: We demonstrated the involvement of PTK7in the progression of thyroid cancer, and therefore provided a novel and promising therapeutic target for thyroid cancer treatment.

[Epidemic trend and control progress of soil-transmitted nematodiasis in Jiangsu Province].

Soil-transmitted nematodiasis was once widely prevalent in Jiangsu Province, which seriously threatened human health and hindered socioeconomic development. The control efforts over decades resulted in a remarkable decline in the prevalence of soil-transmitted nematode human infections in Jiangsu Province, with a reduction from 59.32% in 1989 to 0.12% in 2019, and the human prevalence remains at < 0.5% since 2013. Since 1987, an integrated strategy has been adopted for the control of soil-transmitted nematodiasis in Jiangsu Province; however, the core interventions varies at different stages, which mainly include deworming, water and sanitation service improvement, health education, and monitoring and assessment. The criteria of effective soil-transmitted nematodiasis control had been achieved in all epidemic counties (districts) of Jiangsu Province by 2019. Further actions to strengthen health education and monitoring and implement precision control measures are required to consolidate the achievements of soil-transmitted nematodiasis control and eliminate the harm of soil-transmitted nematodiasis to humans. This review summarizes the epidemiology, control progress and evolution of control strategy of soil-transmitted nematodiasis in Jiangsu Province.

[Establishment of a nucleic acid assay for detection of Echinococcus granulosus based on recombinase-aided isothermal amplification assay].

OBJECTIVE: To establish a nucleic acid assay for detection of Echinococcus granulosus based on recombinase-aided isothermal amplification (RAA) assay. METHODS: The 12S rRNA gene of E. granulosus was selected as the target gene, and the specific primers and fluorescent probes for RAA assay were designed, screened and synthesized to establish a fluorescent RAA assay for detection of E. granulosus. The sensitivity of the fluorescent RAA assay was evaluated using different copy numbers of target gene sequence-contained recombinant plasmids and various concentrations of E. granulosus genomic DNA as templates, and the specificity of the fluorescent RAA assay was evaluated using the genomic DNA from E. granulosus, E. multilocularis, Schistosoma japonicum, S. mansoni, Ancylostoma duodenale, Clonorchis sinensis, Taenia saginata, Spirometra mansoni and Taenia solium as templates. RESULTS: A fluorescent RAA assay was successfully established for detection of E. granulosus, which achieved specific amplification of E. granulosus genomic DNA within 20 min at 39 ℃. The lowest detection limit of the fluorescent RAA assay was 10 copies/µL of recombinant plasmids and 0.1 ng/µL E. granulosus genomic DNA, which exhibited a high sensitivity, and the fluorescent RAA assay was all negative for the genomic DNA from E. multilocularis, S. japonicum, S. mansoni, A. duodenale, C. sinensis, T. saginata, Spirometra mansoni and T. solium, which exhibited a high specificity. In addition, this fluorescent RAA assay successfully detected genomic DNA from E. granulosus cysts. CONCLUSIONS: A rapid, sensitive and specific fluorescent RAA assay is successfully established for nucleic acid detection of E. granulosus.

[Establishment and evaluation of a novel DNA detection method based on recombinase-aided isothermal amplification assay for Giardia lamblia].

OBJECTIVE: To establish a novel nucleic acid assay for detection of Giardia lamblia based on the recombinase-aided isothermal amplification (RAA) assay, and evaluate its sensitivity and specificity for detection of G. lamblia. METHODS: The specific primer sequences and florescent probes were designed and synthesized based on the G. lamblia ß-giardin gene as the target gene, and a fluorescent RAA assay was established. The recombinant plasmids at various copies (containing the ß-giardin gene target sequence) and the genomic DNA of G. lamblia at various concentrations were used as templates for the fluorescent RAA assay to assess the sensitivity, and the genomic DNA from G. lamblia, Schistosoma japonicum, Clonorchis sinensis, Cryptosporidium parvum, Ascaris lumbricoides, Salmonella and Shigella was used as templates to assess the specificity of the fluorescent RAA assay. RESULTS: A novel fluorescent RAA assay was successfully established for detection of G. lamblia, which allowed the rapid and specific amplification of the target gene fragments at 39 ℃ within 20 min. The sensitivities of the fluorescent RAA assay were 102 copies/µL and 1 pg/µL for detection of the recombinant plasmid and G. lamblia genomic DNA, respectively, and the fluorescent RAA assay was negative for detection of the genomic DNA from S. japonicum, C. sinensis, C. parvum, A. lumbricoides, Salmonella and Shigella, which showed a high specificity. CONCLUSIONS: A fluorescent RAA assay, which is simple, sensitive and specific, is successfully established for nucleic acid detection of G. lamblia.

[Investigation on the risk of of Anisakis infection among high - risk populations along the coastal areas of Jiangsu Province].

OBJECTIVE: To investigate the risk of Anisakis infections among high-risk populations along the coastal areas of Jiangsu Province, so as to develop the strategy for the prevention and control of anisakiasis in the province. METHODS: Three counties along the coastal areas of Jiangsu Province were selected as the study sites in 2018, including Rudong County in Nantong City, Haizhou District in Lianyungang City and Dongtai City in Yancheng City. The knowledge, attitude and practice (KAP) of anisakiasis prevention and control, and the prevalence of serum specific IgG antibody against Anisakis were investigated among high-risk populations among these three study sites, including fishermen, fish seller and people who liked eating fresh and live marine fish. Factors affecting the prevalence of the specific IgG antibody against Anisakis were identified using a multiple logistic regression model. In addition, Anisakis larvae infections were detected in fresh and live marine fish samples collected from local markets, and the prevalence and intensity of Anisakis infections were estimated. RESULTS: A total of 625 high-risk populations were investigated, including 349 men (55.8%). Only 13.0% of the subjects heard about anisakiasis, and a low awareness rate of anisakiasis prevention and control knowledge was seen among these three types of high-risk populations. There were 21.6% of the subjects eating raw or half-cooked marine fish, 5.8% eating undercooked marine fish, 3.2% presenting vomiting, nausea and diarrhea after eating marine fish, 5.1% developing systemic allergic symptoms, and 65.6% using the same chopping board for raw and cooked food. The sero-prevalence of the anti-Anisakis IgG antibody was 7.0% among the study subjects. Multiple logistic regression analysis identified education level [OR = 0.687, 95% CI (0.478, 0.987)] and development of systemic allergic symptoms [OR = 4.641, 95% CI(1.411, 15.268)]as factors affecting the positive anti-Anisakis IgG antibody among the study subjects. Among 494 fresh and live marine fish detected, the prevalence and intensity of Anisakis larvae infection was 64.0% and 8.1 larvae per fish, with high prevalence seen in Trichiurus haumela and Pneumatophorus japonicas. CONCLUSIONS: The awareness of anisakiasis prevention and control knowledge is low among the high-risk populations living along the coastal areas of Jiangsu Province, and there are high-risk behaviors, such as eating raw or half-cooked food, using the same chopping board for raw and cooked food. In addition, the prevalence of Anisakis infections is high in the marine fish in these areas. Therefore, the health education and health promotion for anisakiasis prevention and control should be intensified.

A novel mechanistic model for nitrogen removal in algal-bacterial photo sequencing batch reactors.

A comprehensive mathematical model was constructed to evaluate the complex substrate and microbial interaction in algal-bacterial photo sequencing batch reactors (PSBR). The kinetics of metabolite, growth and endogenous respiration of ammonia oxidizing bacteria, nitrite oxidizing bacteria and heterotrophic bacteria were coupled to those of microalgae and then embedded into widely-used activated sludge model series. The impact of light intensity was considered for microalgae growth, while the effect of inorganic carbon was considered for each microorganism. The integrated model framework was assessed using experimental data from algal-bacterial consortia performing sidestream nitritation/denitritation. The validity of the model was further evaluated based on dataset from PSBR performing mainstream nitrification. The developed model could satisfactorily capture the dynamics of microbial populations and substrates under different operational conditions (i.e. feeding, carbon dosing and illuminating mode, light intensity, influent ammonium concentration), which might serve as a powerful tool for optimizing the novel algal-bacterial nitrogen removal processes.