BACKGROUND: 3D-printing is widely used in regenerative medicine and is expected to achieve vaginal morphological restoration and true functional reconstruction. Mesenchymal stem cells-derived exosomes (MSCs-Exos) were applyed in the regeneration of various tissues. The current study aimed to explore the effctive of MSCs-Exos in vaginal reconstruction. METHODS: In this work, hydrogel was designed using decellularized extracellular matrix (dECM) and gelatin methacrylate (GelMA) and silk fibroin (SF). The biological scaffolds were constructed using desktop-stereolithography.The physicochemical properties of the hydrogels were evaluated; Some experiments have been conducted to evaluate exosomes’ effect of promotion vaginal reconstruction and to explore the mechanism in this process. RESULTS: It was observed that the sustained release property of exosomes in the hydrogel both in vitro and in vitro.The results revealed that 3D scaffold encapsulating exosomes expressed significant effects on the vascularization and musule regeneration of the regenerative vagina tissue. Also, MSCs-Exos strongly promoted vascularization in the vaginal reconstruction of rats, which may through the PI3K/AKT signaling pathway. CONCLUSION The use of exosome-hydrogel composites improved the epithelial regeneration of vaginal tissue, increased angiogenesis, and promoted smooth muscle tissue regeneration. 3D-printed, lumenal scaffold encapsulating exosomes might be used as a cell-free alternative treatment strategy for vaginal reconstruction.

Objective:To investigate the incidence and clinical characteristics of brucellosis in Yantai City of Shandong Province, and to provide reference for formulating prevention and control strategies of brucellosis.Methods:From January 2017 to December 2019, brucellosis cases reported by the Infectious Disease Reporting Information Management System of China Disease Prevention and Control Information System in Yantai City were analyzed, and their risk factors and clinical characteristics were analyzed.Results:A total of 225 cases of brucellosis were reported in Yantai City from 2017 to 2019, including 138 males and 87 females, with a gender ratio of 1.59 ∶ 1.00; the age was (43.25 ± 8.61) years, and the most common age was 30 - < 40 years (80 cases); in the regional distribution, Zhifu District was the most (96 cases); the onset time was mainly from January to May (147 cases). Among the risk factors of brucellosis cases, 196 cases (87.11%, 196/225) had close contact with animals or their products. The main clinical characteristics of brucellosis cases were fever (203 cases), fatigue (188 cases), arthralgia (184 cases) and hyperhidrosis (148 cases). All 225 cases of brucellosis were diagnosed in laboratory, including 56 cases (24.89%) with titer 1 ∶ 100 of serum tube agglutination test; 75 cases (33.33%) with titer 1 ∶ 200; and 94 cases (41.78%) with titer 1 ∶ 400. With the increase of titer level, the incidence of fever, hyperhidrosis, arthralgia and fatigue increased, and the differences were statistically significant ( P < 0.05). Conclusions:The incidence of brucellosis in Yantai City is mainly young and middle-aged males, which is common in people who have contact history with animals or their products, and the clinical manifestations are lack of specificity. Health education should be carried out among high-risk groups to improve their self-protection awareness, and brucellosis surveillance should be strengthened to reduce the incidence of brucellosis.

Objective:To analyze the incidence characteristics of occupational chronic benzene poisoning under two diagnostic criteria.Methods:In March 2020, 126 patients who were divided into the old criteria group (74 cases) and the new criteria group (52 cases) were retrospectively analyzed. These patients were diagnosed with occupational chronic benzene poisoning, and were diagnosed in our hospital during the period of January 2009 to December 2019. The gender composition, age of onset, years of benzene exposure, industry distribution, work type, benzene concentration in working environment and diagnostic grade of the two groups of patients were analyzed and compared. The follow-up of 22 benzene poisoning observation subjects under the old criteria were retrospectively analyzed.Results:There were no statistically significant differences in gender composition, age of onset, years of benzene exposure, industry distribution and work type between the old criteria group and the new criteria group ( P>0.05) . In the old criteria group and the new criteria group, 41.9% (31/74) and 17.3% (9/52) of the patients' workplace benzene concentration exceeded the maximum allowable concentration, respectively. The composition of different benzene concentration in the workplace between the two groups showed statistically significant ( P<0.05) . In the old criteria group, the proportion of mild poisoning (79.7%, 59/74) was the majority, while in the new criteria group, the proportion of moderate and severe poisoning (51.9%, 27/52) were the majority, and there was statistically significance in the composition ratio of diagnostic grade between the two groups ( P<0.05) . Under the old criteria, after folow-up of 22 cases of benzene poisoning observation subjects, we observed that 8 cases (36.4%) progressed to the level of chronic benzene poisoning. Conclusion:The revision of diagnostic criteria for benzene poisoning may affect the composition of diagnosis classification. Based on the rights and interests of workers, formulating more complete diagnostic criteria and system policies will be more conducive to the development of occupational benzene poisoning prevention and control.

Objective:To analyze the incidence characteristics of occupational chronic benzene poisoning under two diagnostic criteria.Methods:In March 2020, 126 patients who were divided into the old criteria group (74 cases) and the new criteria group (52 cases) were retrospectively analyzed. These patients were diagnosed with occupational chronic benzene poisoning, and were diagnosed in our hospital during the period of January 2009 to December 2019. The gender composition, age of onset, years of benzene exposure, industry distribution, work type, benzene concentration in working environment and diagnostic grade of the two groups of patients were analyzed and compared. The follow-up of 22 benzene poisoning observation subjects under the old criteria were retrospectively analyzed.Results:There were no statistically significant differences in gender composition, age of onset, years of benzene exposure, industry distribution and work type between the old criteria group and the new criteria group ( P>0.05) . In the old criteria group and the new criteria group, 41.9% (31/74) and 17.3% (9/52) of the patients' workplace benzene concentration exceeded the maximum allowable concentration, respectively. The composition of different benzene concentration in the workplace between the two groups showed statistically significant ( P<0.05) . In the old criteria group, the proportion of mild poisoning (79.7%, 59/74) was the majority, while in the new criteria group, the proportion of moderate and severe poisoning (51.9%, 27/52) were the majority, and there was statistically significance in the composition ratio of diagnostic grade between the two groups ( P<0.05) . Under the old criteria, after folow-up of 22 cases of benzene poisoning observation subjects, we observed that 8 cases (36.4%) progressed to the level of chronic benzene poisoning. Conclusion:The revision of diagnostic criteria for benzene poisoning may affect the composition of diagnosis classification. Based on the rights and interests of workers, formulating more complete diagnostic criteria and system policies will be more conducive to the development of occupational benzene poisoning prevention and control.

Objective: To study the effect of apoptosis-stimulating protein 2 of p53 (ASPP2) on the activation and apoptosis of hepatic stellate cells induced by transforming growth factor-β1 (TGF - β1), and to explore the role of autophagy in this process. Methods: Mouse hepatic stellate cells were primarily isolated and cultured with green fluorescent protein (GFP) expressing empty vector adenovirus (Ad-GFP) and ASPP2 expressing adenovirus (Ad-ASPP2) for 12 h by transfection kit, and then treated with TGF-β1 (10ng/ml) for 24 h. The experiments were grouped as follows: control group: green fluorescent protein (GFP) expressing empty vector adeno (Ad-GFP); experimental group 1: transfected with Ad-GFP and added with TGF-β1; experimental group 2: transfected with Ad-ASPP2 and induced by TGF-β1. Western blot and quantitative fluorescence PCR were used to detect the expression of ASPP2, α-smooth muscle actin (SMA). At the same time, autophagy was determined by microtubule-associated protein 1 light chain 3-β (LC3). Autophagy and apoptosis of MHSc were observed by immunocytochemistry and RNA interference (RNAi). Multiple pairwise-comparisons between the mean of groups was performed by one-way ANOVA. Results: The relative expression of α-SMA mRNA in mHSC of TGF-β1 + Ad-GFP group (16.83 ± 2.41) was significantly higher than Ad-GFP group (3.62 ± 0.56) (P < 0.05), while the relative expression of α-SMA mRNA (4.22 ± 0.48) in TGF-β1 + Ad-GFP group was significantly lower than TGF-β1 + Ad-GFP group (P < 0.05). The expression of α-SMA protein in each group was consistent with mRNA expression. The proportion of mHSC autophagy in TGF-β1 + Ad-GFP group (80%) was significantly higher than Ad-GFP group (35%); however, there was no statistically significant difference between the two groups. The proportion of mHSC autophagy in TGF-β1 + Ad-ASPP2 group was 42%, which was significantly lower than TGF-β1 + Ad-GFP group, but the apoptotic rate was significantly increased. Cells were simultaneously treated with autophagy inhibitors 3-MA and TGF-β1. The level of autophagy was not statistically significantly different from that of TGF-β1 + Ad-ASPP2 group, but the apoptotic rate was increased. In addition, the RNAi group added with ASPP2 had increased autophagy (LC3-II/LC3-I) than control RNAi group, and the rate of apoptosis was significantly decreased. Conclusion Overexpression of ASPP2 can alleviate the activation of mHSC and promote the apoptosis of HSC by inhibiting autophagy, so as to alleviate liver fibrosis.

Objective To evaluate the differences of the malignant rate of thyroid nodules between primary hyperparathyroidism (PHPT) patients and health examination population, and analyze the clinical characteristics of PHPT combined with papillary thyroid carcinoma (PTC). Methods The clinical data of 158 PHPT patients who had underwent parathyroidectomy and 468 thyroid nodule patients who had underwent fine-needle aspiration (FNA) from 2009 to 2014 were retrospectively analyzed, and the detection rate of PTC in patients with thyroid nodule was recorded. Results Of the 158 patients with PHPT, 61 patients had thyroid nodules, the incidence of PHPT with thyroid nodule was 38.6% (61/158), and postoperative pathologically proved PTC was in 12 cases (19.7%). Of the 468 thyroid nodule patients who had underwent FNA, postoperative pathologically proved PTC was in 19 patients (4.1% ). The malignant rate of thyroid nodules in PHPT patients was significantly higher than that in health examination population (P＜0.01). The adjusted serum calcium in patients with PHPT combined with PCT was significantly lower than that in patients with PHPT combined with benign thyroid nodules and patients with PHPT without thyroid nodules: (2.63 ± 0.26) mmol/L vs. (2.92 ± 0.32) and (2.93 ± 0.47) mmol/L, and there was statistical difference (P＜0.05). Conclusions The malignant rate of thyroid nodules in PHPT patients is significantly higher than that in patients from health examination population, which suggests that PHPT might be a risk factor for the malignancy of thyroid nodules. A lower level of serum calcium may predict the existence of PCT in PHPT patients with thyroid nodules.

Objective To study the effect of radiofrequency ablation (RFA) with sublethal temperature on the production of liver cancer stem cells (LCSCs) and on the expression of LCSCs-related transcriptional factors.Methods Mouse hepl-6 hepatoma cell line and clinical samples of patients with hepatocellular carcinoma (HCC) were used to test the expressions of LCSCs-related markers and transcriptional factors.Results Different temperatures were used to stimulate Hep1-6 cells,and it was proved that the temperature of 45℃ was a sublethal temperature that could not induce cell death.Flow cytometry testing showed that treatment with 45℃ could obviously increase CD13+,CD44+,CD90 and CD133+ Hep1-6 cells,suggesting that treatment with 45℃ could increase the production of above mentioned types of LCSCs in hep1-6 cells.Real-time quantitative polymerase chain reaction (RT-qPCR) assay indicated that the temperature of 45℃could cause significant increase in CD13,CD90 and CD133 mRNA.In all 5 HCC patients,CD13 mRNA in the recurrent HCC lesions was remarkably increased,CD133 mRNA was increased in 4 patients with recurrent HCC,and CD90 mRNA was increased in only one patient with recurrent HCC.Flow cytometry testing revealed that CD13+ LCSCs were strikingly increased in 4 recurrent HCC patients,while CD133+LCSC was increased in only one patient,suggesting that more close correlation existed between the increase of CD13+ LCSCs and the temperature of 45℃.RT-qPCR assay showed that in 4 recurrent HCC patients with increased CD13+ LCSC,the Sox2 and Stat2 among 13 LCSCs-related transcriptional factors were obviously increased.Flow cytometry testing showed that 45℃ treatment also increased the expression of Sox2 and Stat1 mRNA in Hep1-6 cells.Finally,Sox2 and Stat1 could be knockdown by siRNAs,indicating that both Sox2 and Stat1 transcriptional factors were involved in 45℃-induced production of CD13+ LCSCs in Hep1-6 cells.Conclusion In RFA therapy,the use of sublethal temperature of 45℃ can increase CD13+LCSCs,which is related to the promotion of Sox2 and Stat1 expression.The results of this study can be used for reference in the research of liver cancer recurrence.

Objective To investigate the effect and underlying mechanisms of ghrelin in a rat model of renal fibrosis. Methods Male Sprague-Dawley rats were divided into 4 groups , including sham operation +saline or brain gut peptide treatment group , model + saline or brain gut peptide treatment group. Unilateral ureteral obstruction (UUO) was established by left ureteral ligation. 7 days and 14 days after operation, the rats were sacrificed , while the kidney tissue of obstruction side was harvested for pathlogical changes through Masson coloration. Expression of α-smooth muscle actin (α-SMA), transforming growth factor beta1 (TGF-β1) and phosphorylated Smad3 (p-Smad3) in renal tissues were analyzed through immunohistochemistry. Expression of α-SMA and TGF-β1 mRNA was detected by real-time-PCR. Apoptosis kidneys cells were marked with TUNEL. Results Ghrelin inhibited renal fibrosis by reducing the production of collagen , restraining extracellular matrix (ECM) deposition and decreasing the expression of α-SMA. Meanwhile, ghrelin inhibited the accumulation of myofibroblasts by blocking the transforming growth factor-β1/Smad3 (TGF-β1/Smad3) signaling pathway. Moreover, ghrelin could attenuate renal tubular cell apoptosis induced by UUO injury. Conclusion Ghrelin can reduce renal fibrosis and renal cell apoptosis induced by UUO , demonstrating that ghrelin is a potent antifibrotic agent that may have therapeutic potential for patients with obstructive nephropathy.

OBJECTIVETo evaluate the clinical value of serum 1,3-beta-D-glucan (BG) and galactomannan (GM) detection for early diagnosis of invasive aspergillosis (IA) in patients after renal transplantation.

METHODSBlood samples collected from 69 renal transplant recipients were divided into diagnosis group, clinical diagnosis group, suspected diagnosis group, and non-infected group for detection of serum BG and GM.

RESULTSThe mean serum levels of BG in the diagnosis group, clinical diagnosis group, and suspected diagnosis group were significantly higher than that in non-infected group (P<0.05). The sensitivity, specificity, and positive and negative predictive values of BG was 69.49%, 70%, 93.18% and 35.71% for IA diagnosis, respectively. The serum levels of GM in the 3 diagnosis groups were also significantly higher than that in the non-infected group (P<0.05) with the sensitivity, specificity, and positive and negative predictive values of 84.75%, 90%, 96.15% and 52.63% for IA diagnosis, respectively.

CONCLUSIONIncreased serum BG and GM levels can serve as the evidence for early diagnosis of IA with a high diagnostic sensitivity and specificity in renal transplant recipients.

Aspergillosis ; diagnosis ; Early Diagnosis ; Humans ; Kidney Transplantation ; Mannans ; blood ; Sensitivity and Specificity ; beta-Glucans ; blood

OBJECTIVETo investigate the protective mechanism of endoplasmic reticulum stress (ERS) inhibition against inflammation-induced acute liver injury using a mouse model.

METHODSMarrow-derived stem cells were isolated from mouse femur and used to derive macrophages for analysis in experimental inflammation conditions, established by exposure to LPS and consequent activation of TLR4. Tunicamycin, an ERS chemical inducer, was applied to interfere the inflammation model condition.Affect on the inflammation-related factor MAPK was detected by western blot, and affects on gene expression of inflammatory factors were measured by real-time quantitative PCR. Affect on TNFa was also measured by ELISA.

RESULTSExpression of TNFa, IL-6 and IL-1b was induced upon exposure to LPS, with the peak levels being reached at 4 hours of exposure (TNFa, 0.82+/-0.24; IL-1 b, 2.20+/-0.69; IL-6, 0.330+/-0.150). Tunicamycin significantly enhanced the LPS-induced up-regulation of TNFa, IL-6 and IL-1b expression (TNFa, 1.44+/-0.38, t=2.8, P<0.05; IL-1b, 16.063.40, t =7.93, P<0.05; IL-6, 31.1610.60, t=5.08, P<0.05). The tuniamycin treatment also enhanced the LPS-induced up-regulation of the protein expression of phospo-p38, phospho-JNK and phoshpo-ERK.

CONCLUSIONERS collaborates with LPS to promote the TLR4-mediated inflammatory response of macrophages, and this collaboration may be a pathogenic mechanism underlying progressive development of acute liver injury.

Animals ; Disease Models, Animal ; Endoplasmic Reticulum Stress ; Inflammation ; Interleukin-6 ; Lipopolysaccharides ; Liver ; Macrophages ; Mice ; Toll-Like Receptor 4 ; Up-Regulation