Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate.

Objective:To prepare a novel ginseng polypeptide thermosensitive hydrogel,and to investigate its repair effect on heat-induced skin injury in the rats and explore the underlying mechanisms.Methods:Thermosensitive hydrogels were formulated using Pluronic F127 and β-sodium glycerophosphate(β-GP),and their phase transition temperatures,spatial structures,elemental compositions,and water retention capacities were evaluated.The rat models of heat-induced skin injury were established and the model rats were divided into PBS group,Gel group,and ginseng polypeptide gel(GP-Gel)group.After 11 d of treatment,the morphological changes of wound and collagen deposition in the wound of the rats in various groups were observed by HE and Masson staining.Immunohistochemistry was used to detect the expression levels of α-smooth muscle actin(α-SMA),connective tissue growth factor(CTGF),basic fibroblast growth factor(bFGF),proliferating cell nuclear antigen(PCNA),cell proliferation marker Ki67,epidermal growth factor(EGF),CD31,vascular endothelial growth factor(VEGF),P50 and P65 proteins in the skin wound tissue of the rats in various groups.Western blotting method was used to detect the expression levels of Toll-like receptor 4(TLR4)in the skin wound tissue of the rats in various groups.ELISA method was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-15(IL-15),and interleukin-10(IL-10)in the serum of the rats in various groups.Results:Compared with PBS and Gel groups,the wound area of the rats in GP-Gel group was reduced(P＜0.01),the expression levels of PCNA,Ki67,EGF,CD31,VEGF,α-SMA,and CTGF proteins in the skin wound tissue were increased(P＜0.05 or P＜0.01),and the expression levels of P65 and TLR4 proteins were decreased(P＜0.01);the level of anti-inflammatory factor IL-10 in serum was increased(P＜0.01),while the levels of pro-inflammatory factors TNF-α,IL-1β,IL-6 and IL-15 were decreased(P＜0.05 or P＜0.01).Conclusion:The ginseng polypeptide thermosensitive hydrogel promotes the repair of heat-induced skin injury by enhancing cell proliferation,collagen synthesis,angiogenesis,and reducing inflammatory responses.

Objective:To analyze the change characteristics of creatinine level in the early stage of patients with diquat (DQ) poisoning, and to explore the early risk factors and the value of prognosis.Methods:A retrospective analysis was carried out on patients with DQ admitted to the the first affiliated hospital of Zhengzhou University from January 2020 to June 2022. The DQ patients were divided into death group and the survival group according to the 28 days survival status after posioning. The basic data and serum indexes and blood gas analysis of the patients on day 1 (D1), day 3 (D3) and day 5 (D5) were collected. The difference of clinical features between the two groups was analyzed, the variables were screened by multiple logistic regression analysis, and the predictive value of the variables was evaluated by drawing receiver operating characteristic curve (ROC curve).Results:A total of 88 patients were included, including 40 patients in the survival group and 48 patients in the death group. The toxic dose in death group was significantly higher than that in survival group [100(40.00, 120.00) mL vs. 50.00(20.00, 90.00) mL, P=0.003]. The higher the toxic dose, the higher the fatality rate. All 4 patients with oral doses greater than 200 mL died. Compared with the survival group, the levels of alanine aminotransferase (ALT) (D3, D5), creatinine (CR) (D3, D5), blood amylase (AMY) (D5) and oxygen partial pressure (PaO 2) (D5) in the death group were significantly higher than those in the survival group (all P<0.05). Multiple Logistic regression analysis showed that CR (D3) and AMY(D5) were independent risk factors for death after poisoning, and PaO 2(D5) was independent protective factor. ROC curve showed that the areas under ROC curve of CR (D3), AMY (D5) and PaO 2 (D5) were 0.814, 0.741 and 0.702, respectively. Conclusion:The higher the oral dose, the higher the death rate. After admission, CR(D3), AMY (D5) and PaO 2 (D5) were independent factors influencing the prognosis of DQ poisoning. In particular, CR (D3) is more effective in predicting death after poisoning.

BACKGROUND:Previous research by the research team found that domestically produced porous tantalum is beneficial for early adhesion and proliferation of MG63 cells,and can be used as a scaffold material for bone tissue engineering. OBJECTIVE:To investigate the effect of domestic porous tantalum modified by osteogenic induction factor slow-release system on the adhesion,proliferation,and differentiation of MG63 cells. METHODS:Osteogenic induction factor slow-release system was constructed by adding 15%volume fraction of osteogenic factor solution to poly(lactic-co-glycolic-acid)gel.The passage 3 MG63 cells were inoculated on a porous tantalum surface(control group),porous tantalum surface coated with poly(lactic-co-glycolic-acid)copolymer gel(gel group),and porous tantalum surface coated with osteoblastic induction factor slow-release system(slow-release system group),and co-cultured for 5 days.The surface cytoskeleton of the material was observed by phalloidine staining.Cell proliferation was detected by flow cytometry.Western blot assay and RT-qPCR were used to detect the protein and mRNA expressions of type Ⅰ collagen,osteopontin,and RUNX-2 on the surface cells of the material. RESULTS AND CONCLUSION:(1)Phalloidine staining showed that MG63 cells adhered to and grew on the surface and inside of the three groups of porous tantalum,and the matrix secreted by the cells covered the surface of the material.(2)Flow cytometry showed that the cell proliferation in the slow-release system group was faster than that in the control group and the gel group(P<0.05).(3)Western blot assay and RT-qPCR showed that the protein and mRNA expressions of type Ⅰ collagen,osteopontin,and RUNX-2 in the slow-release system group were higher than those in the control group and gel group(P<0.05).(4)The results showed that the domestic porous tantalum modified by the osteogenic induction factor slow-release system was beneficial to the adhesion,proliferation,and differentiation of MG63 osteoblasts.

A series of biodegradable nanoparticle-based drug delivery systems have been designed utilizing poly(β-amino ester)-guanidine-phenylboronic acid (PBAE-G) polymers. In this study, a novel Lentinan-Functionalized PBAE-G-nanodiamond system was developed to carry ovalbumin (LNT-PBAE-G-ND@OVA). The impact of this drug delivery system on the activation and maturation of macrophages was then assessed. Furthermore, LNT-PBAE-G-ND@OVA induced potent antibody response and showed no obvious toxicity in vitro and in vivo. Moreover, treatment with LNT-PBAE-G-ND@OVA was sufficient to alter the expression of genes associated with the cGAS-STING pathway, and the LNT-PBAE-G-ND@OVA induced upregulation of costimulatory molecules. LNT-PBAE-G-ND@OVA treatment was sufficient to induce macrophage activation through a complex mechanism in which cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) synthase (cGAS)-stimulator of interferon genes (STING) signaling plays an integral role.

Temporomandibular disorders (TMD) are a heterogeneous group of diseases that affect the temporomandibular joint, chewing muscle system, dental occlusion, and even various structures throughout the body, with significant characteristics of biological-psychological-social pattern. TMD related chronic pain, as the most important clinical symptom, can result in negative emotions seriously affecting patients′ quality of life and physical and mental health. Although a variety of therapies have been previously reported to treat TMD related chronic pain, there is a lack of widely recognized therapies. Professor Jason W Busse (from Michael G DeGroote National Pain Centre, McMaster University, Hamilton ON, Canada) took the lead and collaborated with multiple internationally renowned schools/hospitals of stomatology to develop an international consensus on the management of chronic pain associated with TMD, a clinical practice guideline, which took two years and was published in December 15th, 2023 in a global top journal of clinical research The British Medical Journal. This clinical practice guideline explored the comparative effectiveness of available therapies for chronic pain associated with TMD, conditionally recommended the specific intervention for different treatment or pain relief, proposed a comprehensive, agreed, and standardized clinical practice guideline. This present article describes the methodology and key elements of the clinical practice guideline to help clinicians fully understand and appropriately apply this guidance, which could provide the references for clinical practice of TMD associated chronic pain in China.

Objective:To investigate the effects and molecular mechanism of non-coding RNA-activated DNA damage(NORAD)induced autophagy on oxaliplatin resistance in adenocarcinoma of esophagogastric junction (AEG).Methods:Four pairs of surgical samples of AEG and para-carcinoma normal tissues from patients with advance AEG treated in Anyang Tumor Hospital from January to June 2023 were collected. The expression of NORAD in AEG and para-carcinoma tissues was analyzed by long non-coding RNA microarray chip. The primary tumor cell line of AEG (PDC) was derived from fresh AEG tissues. Oxaliplatin-resistant cell lines of PDC and AEG cell line OE19 (PDC-R and OE19-R) were established. NORAD expression knockdown PDC-R and OE19 cell lines (shNORAD PDC-R and shNORAD OE19-R) were prepared by transfection. The target of NORAD, the correlation and interaction between microRNA-433-3p (miR-433-3p) and NORAD were predicted using Starbase v3.0 and DIANA-lncBase v3.0. PDC, PDC-R, OE19 and OE19-R cells were co-transfected with miR-144-3p and wild-type NORAD (NORAD-WT) or mutant NORAD (NORAD-Mut) plasmid, respectively. Dual-luciferase reporter assay was used to verify the correlation between NORAD and miR-433-3p. The expression levels of NORAD and miR-433-3p in normal gastric mucosal cell line GES-1 and AEG cell lines PDC, PDC-R, shNORAD PDC-R, OE19, OE19-R and shNORAD OE19-R were detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of p62 protein and microtubule-associated protein 1 light chain 3B-Ⅱ (LC3B-Ⅱ) was determined by Western blotting. The half inhibitory concentration (IC 50) of PDC, PDC-R, shNORAD PDC-R, OE19, OE19-R and shNORAD OE19-R cells was measured by cell counting kit-8 (CCK-8) assay. Independent sample t-test was used for statistical analysis. Results:The results of microarray analysis showed that NORAD was significantly up-regulated in AEG compared with that in para-carcinoma tissues (fold change≥2.0, P<0.05). Bioinformatics studies found that miR-433-3p was the potential target of NORAD. The results of dual-luciferase reporter assay indicated that the relative luciferase activity of the NORAD-WT group was lower than that of NORAD-Mut group in PDC and PDC-R cells (0.441±0.104 vs. 0.928±0.204, 0.449±0.112 vs. 0.947±0.201), and the differences were statistically significant ( t=-14.74 and -14.94, both P<0.001). The results of dual-luciferase reporter assay of OE19 and OE19-R cell lines were the same as those of PDC cell lines. The results of qRT-PCR showed that the expression of NORAD in GES-1 cells (1.016±0.213) was lower than that of PDC cells (2.194±0.322) and PDC-R cells (4.040±0.336), and the differences were statistically significant ( t=-14.94 and -37.21, both P<0.001). Furthermore, the expression level of NORAD in PDC was also found to be lower than that in PDC-R cells, and the difference was statistically significant ( t=-19.43, P<0.001). Additionally, shNORAD PDC-R cells exhibited lower expression level of NORAD (0.290±0.165) compared with PDC-R cells, and the difference was statistically significant ( t=-49.05, P<0.001). The expression level of miR-433-3p in GES-1 cells (1.017±0.248) was higher than that in PDC cells (0.470±0.156) and PDC-R cells (0.203±0.045), and the differences were statistically significant ( t=9.15 and 15.85, both P<0.001). Moreover, the expression level of miR-433-3p was found to be higher in PDC cells compared with PDC-R cells, and the difference was statistically significant ( t=8.11, P<0.001). Additionally, the expression level of miR-433-3p in shNORAD PDC-R cells (0.699±0.256) was also higher than that in PDC-R cells ( t=9.37, P<0.001). The results of Western blotting showed that the expression of LC3B-Ⅱ in PDC-R was higher than that in PDC cells (0.426±0.060 vs. 0.212±0.041), the expression of LC3B-Ⅱ in shNORAD PDC-R cells (0.155±0.029) was lower than that in PDC cells, and the differences were statistically significant ( t=8.70 and -79.45, both P<0.001). However the expression of p62 protein in each cell line showed an opposite trend, with a lower relative expression in PDC-R than PDC (0.205±0.031 vs. 0.311±0.400), and the expression in shNORAD PDC-R (0.504±0.084) was higher than that in PDC, and the differences were statistically significant ( t=-31.19 and 62.80, both P<0.001). The expression patterns of NORAD, miR-433-3p, LC3B-Ⅱ and p62 proteins in OE19, OE19-R and shNORAD OE19-R cells were similar to those in PDC. The results of CCK-8 assessment of target cell viability showed that the IC 50 values of PDC, PDC-R and shNORAD PDC-R cell lines were 14.28, 22.27 and 2.51 μg/mL, respectively; and the IC 50 values of OE19, OE19-R and shNORAD PDC-R cell lines were 3.95, 8.12 and 1.89 μg/mL, respectively. Conclusions:NORAD is highly expressed in AEG tissues and cells. NORAD is overexpressed in oxaliplatin-resistant cell lines and increase the autophagy activity of cells. After NORAD is knockdown, autophagy activity is inhibited and the sensitivity of AEG cells to oxaliplatin is significantly enhanced.

Objective:To investigate the influencing factors of secondary osteoporosis in stroke patients with hemiplegia, and to construct a nomogram prediction model and evaluate it.Methods:The study subjects were 110 patients with hemiplegia after stroke who were treated in our hospital from Jun. 2019 to Jun. 2023, and were divided into osteoporosis group and non-osteoporosis group by bone mineral density detection. Clinical data and laboratory indicators were collected. Single factor analysis and binary Logistic multiple factor regression analysis were used to screen the influencing factors. R software (R3.3.2) and software package rms were used to construct the nomogram prediction model.Results:There were 52 patients with osteoporosis and 58 patients without osteoporosis. In the osteoporosis group, the proportion of female, unilateral anterior circulation multiple infarction and less sunlight was significantly higher than that in the non-osteoporosis group, with statistical significance ( χ2=8.27, 14.77 and 6.96, respectively, P<0.05). Systolic blood pressure (159.32±21.72 vs. 151.67±19.52), total cholesterol (4.29±0.50 vs. 3.57±0.42), LDL-C (2.87±0.33 vs. 2.04±0.31), Hcy (3.81±2.51 vs. The level of 112.33±2.47 was significantly higher than that of the non-osteoporosis group, and the difference was statistically significant ( t was 5.23, 8.38, 7.98 and 5.63, respectively, P<0.001). The level of albumin (38.15±5.21 vs. 33.26±5.73) was significantly lower than that of the non-osteoporosis group. The difference was statistically significant ( t=4.90, P<0.05). Binary Logistic regression analysis showed that female, higher total cholesterol, LDL-C, Hcy levels and less sun exposure were independent risk factors for secondary osteoporosis in stroke patients with hemiplegia ( P<0.05). ROC curve analysis results showed that the area under the curve of the established model to predict secondary osteoporosis in stroke hemiplegia patients was 0.891 (0.833-0.949), and the sensitivity and specificity were 80.8% and 79.3%, respectively. Conclusion:The categorization and consistency of the nomogram model based on the influencing factors of secondary osteoporosis in patients with stroke hemiplegia are good, which can provide a certain reference for the identification and early intervention of high-risk groups with stroke hemiplegia.

Objective To investigate the value of quantitative CT(QCT)parameters related to abdominal and dorsal fat content for predicting early postoperative complications of esophageal cancer.Methods A total of 184 patients who underwent radical esophageal cancer surgery were retrospectively collected and divided into complication group(n=76)and control group(n=108)according to whether early postoperation complication(within 30 days after surgery)occurred or not.QCT was used to obtain parameters related to abdominal and dorsal fat content,including visceral fat area(VFA),subcutaneous fat area(SFA),VFA/SFA and the degree of muscle fat infiltration(MFI)of posterior vertebral muscles based on L3 central-level CT images.Univariate analysis and multivariate logistic regression were used to analyze clinical and pathological data as well as QCT parameters related to abdominal and dorsal fat content to screen independent risk factors for early postoperative complications of esophageal cancer.Then receiver operating characteristic(ROC)curve was drawn,and the area under the curve(AUC)was calculated to evaluate the efficacy of each independent risk factor alone and their combination for predicting early postoperative complications of esophageal cancer.Results VFA/SFA and MFI degree of posterior vertebral muscles were both independent risk factors for early postoperative complications of esophageal cancer(OR=5.121,1.110,both P＜0.05).The AUC of VFA/SFA and MFI degree of posterior vertebral muscle was 0.81 and 0.77,respectively,while of their combination was 0.84.Conclusion QCT parameters related to abdominal and dorsal fat content could be used to effectively predict early complications of esophageal cancer after surgery.

Objective To explore the feasibility of minimally invasive autopsy by natural orifice transluminal endoscopic surgery.Methods Autopsy was performed on a deceased patient with COVID-19 via transesophageal,transtrachea,and transgastric natural orifice transluminal endoscopic surgery.The white light endoscopic manifestations of the corresponding organs were observed,and organ tissue specimens were obtained for routine pathological examination.Results All four pathways reached the corresponding organs successfully.Diffuse congestion and submucous bleeding were seen in the trachea,bronchus and bronchus of the pulmonary lobes.The bronchus of the left lower lobe was filled with dark red sputum;the surface of the left lung was congested obviously.Four thrombi and plaque rupture were seen on the aortic wall.The gastric mucosa was congested,eroded,and had active ulcers.The surface of heart and liver was smooth.Small lamellar panniculitis was seen in the omentum.Routine pathology showed chronic inflammation with acute inflammation of the bronchial mucosa and inflammatory exudation,and partial squamous metaplasia of the epithelium.In lung tissue,some alveolar epithelial hyperplasia,a little fibrin-like exudation,widened alveolar septa,and infiltration of acute and chronic inflammatory cells were seen.The columnar epithelial mucosa of the gastric mucosa showed chronic inflammation with acute inflammation and exudates and fungal masses.Conclusion Natural orifice transluminal endoscopic surgery is feasible for autopsy,and covid-19 virus can cause multi-system and multi-organ damage.