BACKGROUND:In the field of tissue engineering,decellularized scaffolds are widely used owing to their biocompatibility.Nonetheless,these scaffolds frequently encounter the limitation of inadequate cell infiltration due to their densely structured composition.Using laser micro-patterning technology for structural modification can improve cell adhesion and proliferation.OBJECTIVE:To summarize the research progress of laser microporous decellularized scaffolds in tissue regeneration.METHODS:Related literature was searched in PubMed and CNKI databases.The Chinese and English search terms were"decellularized scaffold,laser micro-patterning technology,laser microporous decellularized scaffold,extracellular matrix,laser drilling,tissue regeneration."References were screened according to inclusion and exclusion criteria,and 65 articles were finally included for analysis and discussion.RESULTS AND CONCLUSION:(1)Laser micro-patterning technology can selectively remove tissues to achieve scaffold modification.Laser treatment can have controllable effects on the pore structure,mechanical properties and cell permeability of the scaffold.(2)To obtain the optimal cell integration and remodeling effect,factors such as scaffold pretreatment,laser and decellularization process sequence,laser parameter setting,decellularization process and pore structure should be considered.(3)At present,the application of laser microporous decellularized scaffolds has shown that it can promote tissue repair,and it is also necessary to solve the problem of tissue thermal damage.Depending on the absorption of different wavelengths of laser light by the tissue,a reasonable selection of laser type is a feasible solution.(4)Tissue damage repair by laser microporous decellularized scaffold is expected to achieve clinical transformation,and optimizing laser technology and carrying out larger scale biosafety testing are future research directions.

BACKGROUND:In the field of tissue engineering,decellularized scaffolds are widely used owing to their biocompatibility.Nonetheless,these scaffolds frequently encounter the limitation of inadequate cell infiltration due to their densely structured composition.Using laser micro-patterning technology for structural modification can improve cell adhesion and proliferation.OBJECTIVE:To summarize the research progress of laser microporous decellularized scaffolds in tissue regeneration.METHODS:Related literature was searched in PubMed and CNKI databases.The Chinese and English search terms were"decellularized scaffold,laser micro-patterning technology,laser microporous decellularized scaffold,extracellular matrix,laser drilling,tissue regeneration."References were screened according to inclusion and exclusion criteria,and 65 articles were finally included for analysis and discussion.RESULTS AND CONCLUSION:(1)Laser micro-patterning technology can selectively remove tissues to achieve scaffold modification.Laser treatment can have controllable effects on the pore structure,mechanical properties and cell permeability of the scaffold.(2)To obtain the optimal cell integration and remodeling effect,factors such as scaffold pretreatment,laser and decellularization process sequence,laser parameter setting,decellularization process and pore structure should be considered.(3)At present,the application of laser microporous decellularized scaffolds has shown that it can promote tissue repair,and it is also necessary to solve the problem of tissue thermal damage.Depending on the absorption of different wavelengths of laser light by the tissue,a reasonable selection of laser type is a feasible solution.(4)Tissue damage repair by laser microporous decellularized scaffold is expected to achieve clinical transformation,and optimizing laser technology and carrying out larger scale biosafety testing are future research directions.

Objective To explore the clinical significance of serum cytokine expression in the hepatitis B surface antigen(HBsAg)sero-clearance in patients with severe hepatitis B.Methods A nested case-control trial was conducted on 14 inpatients with severe hepatitis B admitted in our hospital from 2006 to 2020.Of them,7 patients(aged 36.57±3.15 years)achieved HBsAg sero-clearance within 1 year after the onset of hepatitis B flares(with abrupt rise of ALT level to＞5 times the upper limit of normal during HBV infection)and were assigned into HBsAg clearance group,while the other 7 patients(aged 34.14±2.97 years)only obtained HBsAg decreased less than 1 g within 1 year after the onset(HBsAg non-clearance group).Then,multiplex liquid-chip assay based on Luminex xMAP was used to detect the expression levels of 48 cytokines such as IFN-γ and IL-2 in serum samples of these 14 patients.Results The serum levels IFN-γ,IL-2Ra and SDF-1α were significantly lower in the HBsAg clearance group than the HBsAg non-clearance group(P＜0.05),but no statistical differences were observed in other 39 cytokines between the 2 groups.And there were 5 cytokines having no mutual expression in both groups.The copy number of HBV DNA was positively correlated with serum HGF(r=0.675,P=0.008)and SDF-1α levels(r=0.587,P=0.027),while negatively with IP-10(r=-0.600,P=0.023)and MIG level(r=-0.640,P=0.014).Meanwhile,a positive correlation was found between HBsAg titer and IL12-p70 level(r=0.593,P=0.025),and a negative correlation between HBsAg titer and TNF-α level(r=-0.609,P=0.021).In addition,the serum total bilirubin level was positively correlated with the expression of SCGF-β(r=0.543,P=0.045).Conclusion Three differentially expressed cytokines and some cytokines related to HBV DNA level and HBsAg titer are found,which may provide new insights into the underlying immunological mechanism of HBV virus clearance caused by hepatitis flares.Meanwhile,it also provides potential biomarkers for HBsAg serological clearance in patients with severe hepatitis B.

Objective:To investigate the application of 3.0T multiparametric magnetic resonance imaging (Mp-MRI) prostate imaging-reporting and data system (PI-RADS) V2.1 score combined with prostate-specific antigen density (PSAD) in the diagnosis of prostate cancer (PCa).Methods:The clinical data of 82 patients with suspected PCa who were admitted to Nantong Second People's Hospital from May 2017 to Octorber 2021 were retrospectively analyzed. The 3.0T Mp-MRI PI-RADS V2.1 score, serum PSAD level and pathological diagnosis were obtained from all patients. The 3.0T Mp-MRI PI-RADS V2.1 score and its distribution as well as serum PSAD level between patients with pathologically diagnosed PCa and patients with prostatic hyperplasia (BPH) were compared. The diagnostic efficiency of 3.0T Mp-MRI PI-RADS V2.1 score and serum PSAD level alone and in combination for PCa was analyzed using receiver operating characteristic (ROC) curve, with pathological results as the gold standard.Results:Pathological diagnosis showed that there were 43 cases (52.44%) of PCa and 39 cases (47.56%) of BPH. There was a statistical difference in the distribution of 3.0T Mp-MRI PI-RADS V2.1 score between PCa and BPH patients ( Z = 32.25, P<0.001). The 3.0T Mp-MRI PI-RADS V2.1 score of PCa patients was higher than that of BPH patients [(4.29±0.25) points vs. (2.24±0.11) points, P < 0.001], the serum PSAD level was higher than that of BPH patients [(0.49±0.15) ng·ml -1·cm -3 vs. (0.27±0.08) ng·ml -1·cm -3, P < 0.001]. The ROC curve analysis showed that area under the curve of 3.0T Mp-MRI PI-RADS V2.1 score, serum PSAD level alone and both together for the diagnosis of PCa were 0.766 (95% CI 0.659-0.852, P < 0.001), 0.793 (95% CI 0.689- 0.874, P < 0.001) and 0.816 (95% CI 0.715-0.893, P < 0.001). Conclusions:3.0T Mp-MRI PI-RADS V2.1 score and serum PSAD level are both elevated in PCa patients. They have certain values in the diagnosis of PCa, and the combination of the two has higher diagnostic efficiency.

Objective:To detect the expression levels of miR-3074-5p and its target gene p27 in the placental tissues of pre-eclampsia (PE) patients, and to preliminarily explore the association of miR-3074-5p/p27 pathway in the pathogenesis of PE. Methods:From September 2017 to March 2018, 16 pregnant women with PE (PE group) and 9 normal pregnant women (control group) were enrolled in Obstetrics Department in the Second Hospital of Tianjin Medical University. The placental expression levels of miR-3074-5p, p27, cyclin D1 (CCND1) and cyclin dependent kinase 2 (CDK2) were determined by quantitative PCR (qPCR), Western blotting and immunohistochemistry (IHC) analyses. The dual-luciferase reporter assay was applied to authenticate the directly targeted effect of miR-3074-5p on expression of p27 mRNA. The expression of p27 in the human extravillous trophoblast cell line HTR-8/SVneo was down-regulated by its specific shRNA, and the effects of down-regulated p27 expression on physiological activities of HTR-8/SVneo cells were detected. Results:Compared with control group, the expression levels of miR-3074-5p ( P=0.034) and CCND1 ( P=0.031) protein in the placenta of PE patients were significantly decreased, whereas the placental p27 ( P=0.010) protein expression was significantly increased in PE group. IHC analysis showed that, the signals of p27 and CCND1 proteins were dominantly detected in syncytiotrophoblast. The results of dual-luciferase reporter assay confirmed that miR-3074-5p could inhibit the expression of p27 by directly bound to the 3'UTR of p27 mRNA. After the p27 expression was down-regulated, the proliferative and invasive activities of HTR-8/SVneo cells were significantly enhanced ( P=0.014, P=0.045). Conclusion:miR-3074-5p might participate in the regulation of physiological activities of extravillous trophoblast cells by its targeted inhibition on p27 expression, and the abnormally decreased miR-3074-5p in trophoblast cells might be associated with the pathogenesis of PE by promoting the expression of p27.

Objective:To detect the expression levels of miR-3074-5p and its target gene p27 in the placental tissues of pre-eclampsia (PE) patients, and to preliminarily explore the association of miR-3074-5p/p27 pathway in the pathogenesis of PE. Methods:From September 2017 to March 2018, 16 pregnant women with PE (PE group) and 9 normal pregnant women (control group) were enrolled in Obstetrics Department in the Second Hospital of Tianjin Medical University. The placental expression levels of miR-3074-5p, p27, cyclin D1 (CCND1) and cyclin dependent kinase 2 (CDK2) were determined by quantitative PCR (qPCR), Western blotting and immunohistochemistry (IHC) analyses. The dual-luciferase reporter assay was applied to authenticate the directly targeted effect of miR-3074-5p on expression of p27 mRNA. The expression of p27 in the human extravillous trophoblast cell line HTR-8/SVneo was down-regulated by its specific shRNA, and the effects of down-regulated p27 expression on physiological activities of HTR-8/SVneo cells were detected. Results:Compared with control group, the expression levels of miR-3074-5p ( P=0.034) and CCND1 ( P=0.031) protein in the placenta of PE patients were significantly decreased, whereas the placental p27 ( P=0.010) protein expression was significantly increased in PE group. IHC analysis showed that, the signals of p27 and CCND1 proteins were dominantly detected in syncytiotrophoblast. The results of dual-luciferase reporter assay confirmed that miR-3074-5p could inhibit the expression of p27 by directly bound to the 3'UTR of p27 mRNA. After the p27 expression was down-regulated, the proliferative and invasive activities of HTR-8/SVneo cells were significantly enhanced ( P=0.014, P=0.045). Conclusion:miR-3074-5p might participate in the regulation of physiological activities of extravillous trophoblast cells by its targeted inhibition on p27 expression, and the abnormally decreased miR-3074-5p in trophoblast cells might be associated with the pathogenesis of PE by promoting the expression of p27.

Objective:To investigate a cluster epidemic of COVID-19 after a mass gathering activity in Ningbo of Zhejiang province and analyze the transmission chain and status of infection cases of different generations.Methods:The tracking of all the close contacts of the first COVID-19 case and epidemiological investigation were conducted on January 29, 2020 after a cluster epidemic of COVID-19 related with a Buddhism rally on January 19 (the 1.19 rally) in Ningbo occurred. The swabs of nose/throat of the cases and close contacts were collected and tested for nucleic acids by real-time fluorescence quantitative RT-PCR.Results:From January 26 to February 20, 2020, a total of 67 COVID-19 cases and 15 asymptomatic infection cases related with the 1.19 rally were reported in Ningbo. The initial case was the infection source who infected 29 second generation cases and 4 asymptomatic infection cases, in whom 23 second generation cases and 3 asymptomatic infection cases once took bus with the initial case, the attack rate was 33.82% (23/68) and the infection rate was 38.24% (26/68). The risks of suffering from COVID-19 and being infected were 28.91 times and 26.01 times higher in rally participants taking bus with initial case compared with those taking no bus with initial case. In this epidemic, 37 third+generation cases and 11 related asymptomatic infection cases occurred, the attack rate was 2.88% (37/1 283) and the infection rate was 4.76% (48/1 008). The main transmission routes included vehicle sharing and family transmission.Conclusion:It was a cluster epidemic of COVID-19 caused by a super spreader in a massive rally. The epidemic has been under effective control.

Objective:To describe the basic characteristics of clusters of coronavirus diseases 2019 (COVID-19) cases in Ningbo, Zhejiang province, and evaluate the generation time (Tg) and basic reproduction number ( R0) of COVID-19. Methods:The basic information and onset times of the clusters of COVID-19 cases in Ningbo were investigated, the inter-generational interval of the cases were fitted by using gamma distribution, and the R0 was calculated based on the SEIR model. Results:In the 15 clusters of COVID-19 cases, a total of 52 confirmed cases, 5 cases of nucleic acid-positive asymptomatic cases. The cases occurred from January 23 to February 4, the cases were mainly women. The incubation period was (6.11±3.38) days, and the median was 5 days. The Tg was (6.93±3.70) days. There were no significant differences in Tg between age group<60 years and age group 60 years and above, and between men and women ( P=0.551). According to the Tg calculated in this paper, the R0 of COVID-19 in Ningbo was 3.06 (95 %CI: 2.64- 3.51); according to the reported case transmission interval of 7.5 days in the literature, the R0 was 3.32 (95 %CI: 2.51-9.38). Conclusion:There is no age and gender specific differences in the Tg of clusters of COVID-19 cases in Ningbo, and COVID-19 has high infectivity and spreading power in early phase.

Objective: To investigate the expression change, biological role and action mechanism of long non-coding RNA (lncRNA) LINC00978 in non-small cell lung cancer (NSCLC). Methods: The expression levels of LINC00978 in tumor tissues and serum samples of NSCLC patients were detected by the qRT-PCR. The effects of knockdown and overexpression of LINC00978 on the biological function of A549 cells were determined by the CCK-8, colony formation, Transwell migration and invasion assays. The action mechanisms of LINC00978 in NSCLC were investigated by the flow cytometry, qRT-PCR and western blot, respectively. Results: The expression levels of LINC00978 in the tissues ( t =2.465, P ＜0.05) and serum samples ( t =8.781, P ＜0.01) of NSCLC patients increased. The knockdown of LINC00978 inhibited the proliferation, migration and invasion of A549 cells ( P ＜0.01) and induced cell cycle arrest at G1 phase and apoptosis of A549 cells ( P ＜0.01). The knockdown of LINC00978 downregulated the expression of Cyclin D1 and Bcl-2 , and upregulated the expression of Bax ( P ＜0.05). In addition, the knockdown of LINC00978 inhibited the expression of N-cadherin, Vimentin, Snail, Slug and Twist, and promoted the expression of E-cadherin ( P ＜0.05). The overexpression of LINC00978 had the opposite effect. Conclusion LINC00978 is highly expressed in NSCLC and can promote the occurrence and progression of NSCLC, which may serve as a potential target for the diagnosis and therapy of NSCLC.

Objective: To determine the changed expression levels, biological roles and underlying mechanism of LncSox4 in non-small cell lung cancer (NSCLC), providing novel biomarkers for NSCLC diagnosis and therapy. Methods: QRT-PCR was used to detect the expression of LncSox4 in the tumor tissues of NSCLC patients. Colony formation, cell growth curve, Transwell migration and invasion assays were used to determine the effects of LncSox4 knockdown on A549 cell function, respectively. Flow cytometry was used to determine the effects of LncSox4 on the progression of A549 cell cycle. QRT-PCR and western blot were used to explore the expressions of genes and proteins in epithelial-mesenchymal transition (EMT). Results: The expression of LncSox4 was upregulated significantly in carcinoma tissues of NSCLC compared to the para-carcinoma tissues (t=7.109，P＜0.01). The growth rate of A549 cells slowed down in LncSox4 knockdown group and the number of formed cell colonies was less than that in control group(P＜0.01). LncSox4 knockdown reduced the migration and invasion abilities of A549 cells (P＜0.01) and induced cell cycle arrest at G1 phase(P＜0.01). LncSox4 knockdown downregulated the protein expressions of Cyclin D1, c-Myc, N-cadherin, and Vimentin, while upregulated the expression of E-cadherin in A549 cells. LncSox4 knockdown also decreased the expressions of EMT-related transcription factors including snail, slug and twist. Conclusion The high expression of LncSox4 in NSCLC may promote malignant progression of NSCLC by enhancing cell proliferation, migration and invasion, suggesting that it should be a promising target for diagnosis and therapy of NSCLC.