1.Research status and reflection on animal models of sepsis
Chinese Journal of Preventive Medicine 2024;58(2):171-176
Infectious diseases are the leading cause of death in children. Sepsis is a critical infectious disease that causes death in children globally, with a high morbidity and mortality rate. It poses a serious threat to children′s health. Early diagnosis has become the key to treating severe sepsis. The establishment of animal models of sepsis can help people better diagnose sepsis and take interventions to improve the prognosis of sepsis patients. This study reviews the types, advantages and disadvantages of existing animal models of sepsis and proposes the optimization of these models to provide a reference basis for the selection and optimization of experimental models and the promotion of the "reverse transformation" of sepsis into clinical practice.
2.The diagnostic value of inflammation-related genes in bronchopulmonary dysplasia
Ya WEN ; Junfeng ZHANG ; Shiqiang SHANG
Chinese Journal of Preventive Medicine 2024;58(6):898-904
This study aims to explore the diagnostic value of inflammation-related genes in peripheral blood mononuclear cells in bronchopulmonary dysplasia (BPD). By using bioinformatics analysis, three datasets including GSE32472, GSE125873, and GSE220135, which contain whole-genome expression profile data of 251 neonates, were included. The GSE32472 dataset was used as a training dataset to detect differentially expressed genes between non-BPD and BPD neonates in peripheral blood mononuclear cells. The gene enrichment analysis (GSEA) was used to detect the pathway enrichment of up-regulated genes in BPD newborns. The main regulatory factors analysis (MRA) algorithm was used to filter the main regulatory genes in the inflammation-related pathway (GO:0006954). After obtaining the main regulatory genes, the expression of the main regulatory genes in the GSE32472, GSE125873, and GSE220135 datasets was detected. Through the logistic regression model, risk scoring was conducted for neonates, and the risk scores of non-BPD and BPD neonates were compared. Lastly, the classification performance of the model was evaluated using the area under the curve (AUC). The results showed that compared with non-BPD neonates, there were 486 up-regulated genes and 433 down-regulated genes in the peripheral blood mononuclear cells of BPD neonates. The inflammation-related pathway was highly enriched in the up-regulated genes. Ultimately, phospholipase C beta 1 (PLCB1), nidogen 1 (NID1), serum response factor binding protein 1 (SRFBP1), centrosomal protein 72 (CEP72), excision repair cross complementation group 6 like (ERCC6L), and peptidylprolyl isomerase like 1 (PPIL1) were identified as the main regulatory genes. The prediction model′s calculation formula for risk score was PLCB1×0.26+NID1×0.97+SRFBP1×1.58+CEP72×(-0.36)+ERCC6L×2.14+PPIL1×0.67. The AUCs in the GSE32472 test dataset, GSE125873 dataset, and GSE220135 dataset were 0.88, 0.86, and 0.89, respectively. This prediction model could distinguish between non-BPD and BPD neonates. In conclusion, the prediction model based on inflammation-related pathway genes has a certain diagnostic value for BPD.
3.Research status and reflection on animal models of sepsis
Chinese Journal of Preventive Medicine 2024;58(2):171-176
Infectious diseases are the leading cause of death in children. Sepsis is a critical infectious disease that causes death in children globally, with a high morbidity and mortality rate. It poses a serious threat to children′s health. Early diagnosis has become the key to treating severe sepsis. The establishment of animal models of sepsis can help people better diagnose sepsis and take interventions to improve the prognosis of sepsis patients. This study reviews the types, advantages and disadvantages of existing animal models of sepsis and proposes the optimization of these models to provide a reference basis for the selection and optimization of experimental models and the promotion of the "reverse transformation" of sepsis into clinical practice.
4.The diagnostic value of inflammation-related genes in bronchopulmonary dysplasia
Ya WEN ; Junfeng ZHANG ; Shiqiang SHANG
Chinese Journal of Preventive Medicine 2024;58(6):898-904
This study aims to explore the diagnostic value of inflammation-related genes in peripheral blood mononuclear cells in bronchopulmonary dysplasia (BPD). By using bioinformatics analysis, three datasets including GSE32472, GSE125873, and GSE220135, which contain whole-genome expression profile data of 251 neonates, were included. The GSE32472 dataset was used as a training dataset to detect differentially expressed genes between non-BPD and BPD neonates in peripheral blood mononuclear cells. The gene enrichment analysis (GSEA) was used to detect the pathway enrichment of up-regulated genes in BPD newborns. The main regulatory factors analysis (MRA) algorithm was used to filter the main regulatory genes in the inflammation-related pathway (GO:0006954). After obtaining the main regulatory genes, the expression of the main regulatory genes in the GSE32472, GSE125873, and GSE220135 datasets was detected. Through the logistic regression model, risk scoring was conducted for neonates, and the risk scores of non-BPD and BPD neonates were compared. Lastly, the classification performance of the model was evaluated using the area under the curve (AUC). The results showed that compared with non-BPD neonates, there were 486 up-regulated genes and 433 down-regulated genes in the peripheral blood mononuclear cells of BPD neonates. The inflammation-related pathway was highly enriched in the up-regulated genes. Ultimately, phospholipase C beta 1 (PLCB1), nidogen 1 (NID1), serum response factor binding protein 1 (SRFBP1), centrosomal protein 72 (CEP72), excision repair cross complementation group 6 like (ERCC6L), and peptidylprolyl isomerase like 1 (PPIL1) were identified as the main regulatory genes. The prediction model′s calculation formula for risk score was PLCB1×0.26+NID1×0.97+SRFBP1×1.58+CEP72×(-0.36)+ERCC6L×2.14+PPIL1×0.67. The AUCs in the GSE32472 test dataset, GSE125873 dataset, and GSE220135 dataset were 0.88, 0.86, and 0.89, respectively. This prediction model could distinguish between non-BPD and BPD neonates. In conclusion, the prediction model based on inflammation-related pathway genes has a certain diagnostic value for BPD.
5.Application of MassARRAY genotyping assay in the diagnosis of neonatal genetic metabolic diseases
Ting ZHANG ; Shiqiang SHANG ; Jianbin YANG ; Xuchen ZHOU ; Yu ZHANG ; Rulai YANG ; Xinwen HUANG
Chinese Journal of Laboratory Medicine 2023;46(2):155-162
Objective:To investigate the accuracy, effectiveness and feasibility of MassARRAY genotyping assay in the diagnoses of neonatal genetic metabolic diseases.Methods:This is a retrospective study. From December 2016 to January 2020, newborns were screened by tandem mass spectrometry at the Zhejiang Newborn Screening Center, among which the data of 7 922 suspected positive cases of genetic metabolic diseases were collected. These patients were then tested for the common variants of 27 genetic metabolic diseases by MassARRAY genotyping assay, along with further testing using Sanger or next-generation sequencing used to verify and/or further search for potential variants.Results:A total of 1 408 cases were tested with MassARRAY. Among these, 307 cases were confirmed with certain genetic metabolic diseases. The detection rate of hyperphenylalaninemia was the highest, followed by primary carnitine deficiency, short acyl-coA dehydrogenase deficiency and methylmalonic acidemia. With these cases, the consistency of Sanger sequencing and MassARRAY was 100% (307/307). Another 287 cases were identified as carriers by MassARRAY with a 49.1% (141/287) consistency in reference to Sanger sequencing, mainly involving SLC22A5 and MCCC1 genes. Meanwhile, 50.8% (146/287) of these cases were found to have another variant mainly involving PAH, PTS and ACADS genes. The remaining 814 cases have no variants; 158 cases out of these patients have continuously abnormal amino acids, acyl carnitines, urine organic acid and/or other biochemical indices, and were tested by next-generation sequencing, among which 38% (60/158) were detected with two variants. In this study, a total of 513 patients with genetic metabolic disease were diagnosed, and the detection rate of MassARRAY was 59.8% (307/513). Conclusions:MassARRAY genotyping assay can be used as an early molecular screening method for neonatal genetic metabolic diseases. The detection rate is particularly high in diseases with a high concentration of hotspot variants, such as hyperphenylalaninemia and primary carnitine deficiency. The future application value of MassARRAY should be further improved by continuously optimizing its ability to identify new disease genes and potential variable sites.
6.Sponge forceps assisted threading with Speedbridge technique for the treatment of acute closed Achilles tendon rupture
Lin SHANG ; Fuqiang MA ; Qi LI ; Yalei WANG ; Xiaolong ZHANG ; Shiqiang SUN ; Guanghui JIA ; Xiangyu WANG ; Aiguo WANG
Chinese Journal of Trauma 2023;39(3):259-264
Objective:To explore the outcome of sponge forceps assisted threading with Speedbridge technique for the treatment of acute closed Achilles tendon rupture.Methods:A retrospective case series study was conducted on 20 patients with acute closed Achilles tendon rupture treated in Zhengzhou Orthopedic Hospital from December 2019 to December 2021. There were 18 males and 2 females, with age range of 24-43 years [(29.5±7.6)years]. All patients were with unilateral injury, involving the left side in 13 patients and right side in 7. Examinations revealed a palpable defect in the Achilles tendon and positive Thompson test. A longitudinal incision was made at the medial edge of the ruptured tendon. Three nonabsorbable sutures were passed through the proximal stump with sponge forceps, bypassed the rupture site and fixed directly into the calcaneal bone. The disrupted tendon ends were aligned by the tendon-bundle technique using 4-0 absorbable sutures. The operation time and incision length were documented. The ankle joint range of motion (dorsiflexion/plantar flexion), American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and Achilles tendon total rupture score (ATRS) in the affected and healthy side were compared at 3, 6 and 12 months postoperatively. The wound healing and complications were observed.Results:All patients were followed up for 12-16 months [(13.2±2.5)months]. The operation time was 40-66 minutes [(52.0±10.3)minutes], with the incision length of 3-4 cm [(3.3±0.7)cm]. In the affected side at 3 and 6 months postoperatively, the ankle joint dorsiflexion [(5.6±1.5)°, (10.5±0.2)°] and plantar flexion [(28.4±3.2)°, (33.5±1.5)°] showed statistically significant difference compared with the healthy side (all P<0.05). The ankle joint dorsiflexion [(13.9±0.7)°] and plantar flexion [(38.3±4.4)°] in the affected side were not statistically different from that of the healthy side at 12 months postoperatively (all P>0.05). The AOFAS ankle-hindfoot score was (58.3±5.4)points, (84.9±7.1)points and (91.8±6.3)points at 3, 6 and 12 months postoperatively, showing a gradual rise (all P<0.05). The ATRS was (60.5±4.9)points, (85.5±9.0)points and (93.1±5.7)points at 3, 6 and 12 months postoperatively, showing a gradual rise (all P<0.05). All incisions were healed primarily. No patients had wound infection, nerve injury or re-rupture. Pain at the anchor insertion site occurred in 2 patients at 1 month after operation and relieved after active functional rehabilitation at 4 months after operation. Transient pain at the Achilles tendon insertion occurred in 1 patient at 6 months after operation, and relieved after 2 weeks of oral non-steroidal anti-inflammatory drugs treatment. Conclusion:For acute closed Achilles tendon rupture, sponge forceps assisted threading with Speedbridge technique can attain short operation time, small incision and good functional recovery, with few complications.
7.Research progress in human cytomegalovirus and its association with TGF-β and Smad signaling pathway
Chinese Journal of Microbiology and Immunology 2022;42(6):490-494
Human cytomegalovirus (HCMV) is a common herpes virus found in human and can establish lifelong latency in the hosts. In healthy population, HCMV usually results in asymptomatic latent infection. However, the latent HCMV can be activated and cause many serious diseases and even death in people with low immunity. Transforming growth factor-β (TGF-β) is a powerful regulator of many cellular pathways, playing important roles in inflammatory response and cell differentiation, as well as immunomodulatory role in viral infection. The classic Smad signaling pathway is involved in the regulation of many cell activities, including growth, differentiation and apoptosis. This review summarized the progress in the association of HCMV infection with TGF-β and the Smad signaling pathway.
8.Combined application of pathogen-based tests and host reaction-based tests
Chinese Journal of Laboratory Medicine 2022;45(6):559-562
The occurrence development and progression of infectious diseases involves the two aspects: the immune response of both the pathogens and the hosts. At present, pathogen-based testing is the gold standard and the most commonly used method for the diagnosis of infectious diseases. The purpose of the immune system is to identify and eliminate invading pathogens. Testing based on host reactions has become an effective auxiliary means adjunct of traditional pathogen-based tests, with the potential to improve the accuracy and efficiency of diagnosis. The combined application of host-based tests and pathogen-based tests is a new field worth exploring.
9.Etiological detection and epidemiological analysis of children with influenza-like illness
Jian LIN ; Wei LI ; Chao ZHOU ; Jiaming TANG ; Yajun GUO ; Shiqiang SHANG
Chinese Journal of Laboratory Medicine 2022;45(6):574-580
Objective:To analyze the status and epidemiological characteristics of respiratory virus infection in children with influenza-like illness in outpatient department, and to provide evidence for the prevention and treatment of children in this area.Methods:Nasopharyngeal swab samples were collected from children who attended the fever clinic of The Children′s Hospital, Zhejiang University School of Medicine due to influenza-like illness from July 2021 to March 2022, and six common respiratory virus nucleic acids were detected by reverse transcription-polymerase chain reaction (RT-PCR). The general information of the children was collected and grouped by gender and age (0-<6 months, 6-<12 months, 1-3< year-old, 3-<6 year-old , and ≥6 year-old), and the chi-square test was used for statistical analysis between the groups to explore the epidemic pattern of respiratory viruses.Results:A total of 739 cases (45.9%, 739/1 609) of respiratory viruses were detected from children with influenza-like illness, including 651 cases (40.5%, 651/1 609) of simple infection and 88 cases (5.5%, 88/1 609) of multiple infections. Respiratory syncytial virus (RSV) was detected in 18.6% (300/1 609), followed by influenza B virus (FluB) in 11.9% (192/1 609), adenovirus (ADV) in 8.3% (134/1 609), parainfluenza virus type 3 (PIV-3) in 7.6% (123/1 609), parainfluenza virus type 1 (PIV-1) in 4.9% (79/1 609), and influenza A virus (FluA) in 0.4% (6/1 609). Multiple infections including double or triple infections, with 81(92.0%, 81/88) cases of double infection and the most common being ADV+RSV (22.7%, 20/88) and 7 (8.0%, 7/88) cases of triple infection. There was a significant difference in the virus detection rate between the age groups (χ2=17.078, P=0.002), with the highest virus detection rate in the 3-<6 years of age group (49.7%, 286/575). Among the detection of simple infection, FluB had the highest detection rate in the ≥ 6 years of age group (26.6%, 98/369), and RSV and PIV-1 had the highest detection rate in the 3-<6 years of age group (20.0%, 115/575 and 5.9%, 34/575). The total monthly virus detection rate increased from 26.8% (37/138) in July to 63.0% (58/92) in January, and decreased to 46.1% (106/230) and 26.8% (37/138) in February and March. The detection rate of RSV was the highest from August to November, the detection rate of FluB was the highest from December to March, the detection rate of ADV increased in December and January, and the detection rate of PIV-3 increased from October to December; the detection rate of PIV-1 did not fluctuate significantly, and FluA was sporadically detected. Conclusions:RSV is the main respiratory virus in children with influenza-like illness. Most respiratory viruses are present as single infections. Multiple infections are more common in double infections. FluB, RSV and PIV-1 infections showed certain age distribution characteristics, especially in children over 3 years of age. The epidemic characteristics of respiratory virus infection show that the epidemic gradually peaks from summer to autumn and winter, and turns into an epidemic decline in spring. RSV was relatively prevalent in autumn, FluB was prevalent in winter and spring, ADV and PIV-3 were prevalent to varying degrees in winter, PIV-1 continued to circulate at a low level, and FluA did not present epidemic characteristics.
10.The role of extracellular vesicles in human cytomegalovirus infection
Huamei LI ; Lin HE ; Ran TAO ; Shiqiang SHANG
Chinese Journal of Laboratory Medicine 2022;45(6):656-660
Extracellular vesicles (EV) are membrane structured vesicles containing proteins, lipids and nucleic acids. EVS produced by virus-infected cells are involved in communication between infected and uninfected cells. EV produced by human cytomegalovirus (HCMV) infected cells can promote the transmission and infection of HCMV and escape the host immune response. It can also activate the body′s immune response against HCMV infection. In-depth study of the mechanism and compositional changes of EV produced by HCMV infected cells will contribute to the prevention, diagnosis, and treatment of HCMV. The current research has achieved some results, but they are not deep enough. Future advancements in EV isolation and identification technologies and the reduction of economic costs will contribute to the extensive development and clinical application of the research.

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