Bone Marrow Mesenchymal Stem Cells (BMSCs) Transplantation Ameliorates Joint Severity and Inflammation in Rats with Arthritis

This study analyzed the action of Bone marrow mesenchymal stem cells (BMSCs) transplantation on arthritis rat model. Arthritis rat model was established using bovine type II collagen and CFA. BMSCs phenotype was assessed by flow cytometry and pathological changes was analyzed by H&E staining along with analysis of joint severity by AI score, inflammation by ELISA as well as level of NPY, MMP-2, and MMP-9. The form of passaged BMSCs was spindle shaped with positive expression of CD29 and CD44. The structure of articular cavity in arthritis rats was disordered with infiltration of inflammatory cells which were ameliorated by BMSCs transplantation. In addition, BMSCs treatment also significantly reduced AI value, the level of VEGF, IL-17 and TNF-α as well as decreased RANK/RANKL expression and increased OPG level. In conclusion, BMSCs transplantation ameliorates inflammation and severity in arthritis rats possibly through regulation of RANK/OPG, indicating that it might be used for the treatment of arthritis patients.

In vitro Culture and Morphometry of Porcine Adipose Derived Mesenchymal Stem Cells (pAD-MSCs)

Background: Mesenchymal stem cells are well known for their self-renewal capacity and ability to differentiate into multiple cell lineages. The aim of the study was to develop a simple technique for isolation of mesenchymal stem cells from porcine adipose tissue and to study the morphometric characteristics of porcine mesenchymal stem cells. Methods: Porcine adipose derived mesenchymal stem cells were isolated in vitro by using collagenase type II enzyme. Cell yield and viability of the cells were calculated by using trypan blue exclusion method using Neubauer’s chamber. Characterization of MSCs were done by using specific cell markers. The morphological changes, morphometry were analysed in culture using Leishman’s stain. The cell doubling (CD) and Population doubling time (PDT) were also calculated. Result: The isolated adherent cells start forming colony and demonstrated an elongated, round and spindle like fibroblastic morphology by day 1. Almost 80-90 per cent confluency was attained on day 8-9 after the initial seeding and was reduced to day 3-4 in the subsequent passages. RT-PCR reactions revealed positive expression of mesenchymal stem cell markers CD44, CD73 and negative expression of CD34, a hematopoietic cell surface marker. Immunocytochemistry also revealed positive expression for CD44 and negative for CD34. In morphometric studies, the cell length, nucleus length, cell width and nucleus width were increased between 24 and 48 hours in both P2 and P3.

Correlation between the features of β1 and β3 integrin expression and lymphangitic metastatic spread in nonspecific invasive breast carcinoma based on tumor morphological heterogeneity

Objective. To study β1 and β3 integrin expression in nonspecific invasive breast carcinoma and to find the associations with parameters of tumor morphological heterogeneity and lymphatic dissemination.Material and Methods. Study group comprised 107 patients with breast cancer. Histological type of tumor corresponded to invasive carcinoma of a nonspecific type (invasive ductal carcinoma) in 100% of cases. Patients did not receive any preoperative treatment. In each case, we performed morphological examination of samples of primary tumor and axillary lymph nodes obtained at the surgical stage of treatment (radical mastectomy or sectoral resection of mammary gland with axillary lymphadenectomy). The parameters of β1 and β3 integrin expression in primary tumor tissue were assessed by immunohistochemistry.Results. The study demonstrated that an increase in the degree of malignancy of breast carcinoma was associated with a decrease in the incidence of positive expression of β1 integrin as well as with an increase in the incidence of positive expression of β3 integrin. Metastases in lymph nodes were significantly less frequently detected in the presence of positive expression of β1 integrin in the alveolar and solid structures compared with the cases of absent expression of the marker in similar structures (48%; χ2 = 3.5; p = 0.05 and 48%; χ2 = 4.8; p = 0.02, respectively). Lymphogenic metastasis were detected significantly more often in cases with positive expression of β3 integrin in discrete groups of cells compared with the cases where the expression of study marker in the described structures was absent (47 and 23%, respectively; χ2 = 5.1; p = 0.02).Conclusion. The results of work showed the presence of relationships between the morphological heterogeneity of the tumor and the parameters of β1 and β3 integrin expression in the parenchymal structures of the neoplasm. The study showed the association of described parameters with the frequency of lymphatic dissemination in patients with breast cancer. Obtained data expand and support previously known evidence and suggest the possibility of assessing the markers as potential prognostic factors predicting the course of cancer.

Comparative Effects of Valganciclovir and Ganciclovir on the Congenital Cytomegalovirus Infection and Hearing Loss: A Randomized Controlled Trial

Background: Cytomegalovirus (CMV) is a highly specific herpes virus spreading only from person to person. Valganciclovir (VGCV) and ganciclovir (GCV) are effective in the treatment of neonatal congenital CMV infection. Objectives: This study aimed to compare the curative effects of VGCV and GCV among neonates with CMV infection and evaluate their effects on hearing. Methods: A total of 48 neonates with congenital CMV infection admitted to Huaian Maternal and Child Health Care Hospital, China, were selected from January 2016 to December 2019 and randomly divided into two equal groups of intervention and group (n = 24 each). While the control group received intravenous injection of GCV, the intervention group received oral VGCV. After a 6-week course of treatment, polymerase chain reaction (PCR) was applied to detect the CMV load in both urine and blood. We used the ELISA method to detect the serum CMV-IgM expression level before and after treatment. Moreover, we compared the positive rates of CMV-DNA and CMV-IgM, hyperbilirubinemia, retinitis, hepatosplenomegaly, thrombocytopenia, neutropenia, as well as the results of hearing screening and brainstem auditory evoked potentials (BAEP). Results: Before treatment, there was no statistical difference in blood/urine CMV-DNA expression level and positive expression rates of blood/urine CMV-DNA and CMV-IgM between the groups (P > 0.05). After treatment, blood/urine CMV-DNA expression and the positive expression rates of blood/urine CMV-DNA and CMV-IgM significantly decreased in both groups compared to before treatment (P < 0.05), but there was no statistical significance between the two groups (P > 0.05). Before treatment, there was no significant difference in hearing abnormality rates between the control (50%) and intervention (62.5%) groups (P > 0.05). After treatment, both the control (20.83%) and intervention (29.17%) groups had significantly decreased hearing abnormality rates, and the difference was statistically significant compared with before treatment (P < 0.05), but the difference between the two groups was not statistically significant (P > 0.05). After treatment, the results of comparing BAEP showed that both groups had no statistically significant differences in the number of neonates with normal hearing, mild hearing loss, moderate to severe hearing loss, severe hearing loss, and extremely severe hearing loss (P > 0.05). Before treatment, both groups had no statistically significant differences in the number of neonates with hyperbilirubinemia, retinitis, hepatosplenomegaly, thrombocytopenia, and neutropenia (P > 0.05). After treatment, while the number of neonates with hyperbilirubinemia, retinitis, hepatosplenomegaly, and thrombocytopenia decreased, neutropenia cases increased, and the difference before and after treatment was statistically significant (P < 0.05); however, the difference between the two groups was not statistically significant (P > 0.05). Conclusions: VGCV is similar to GCV in the treatment of neonatal congenital CMV infection, but the oral route of administration of VGCV is more acceptable among neonates.

Segnet Network Algorithm-Based Ultrasound Images in the Diagnosis of Gallbladder Stones Complicated with Gallbladder Carcinoma and the Relationship between P16 Expression with Gallbladder Carcinoma

The study focused on how to improve the diagnostic coincidence rate of patients with gallbladder stones and gallbladder cancer based on an optimized Segnet network algorithm and the relationship of gallbladder cancer with multiple tumor suppressor 1 (P16). 300 patients diagnosed with gallbladder cancer in the hospital were selected as the research subjects. The pyramid pooling operation was incorporated into the original Segnet network algorithm, and its performance was evaluated, factoring into the intersection of union (IoU), algorithm precision (Pre), and recall rate (Recall). After 8 hours of fasting, conventional ultrasound and contrast-enhanced ultrasound examinations were performed, and the images were evaluated by three experienced ultrasound diagnosticians. The positive signal of P16 immunohistochemical staining was brownish yellow, which was generally concentrated in the nucleus, and a small part was located in the cytoplasm. In each slice, ten visual fields were selected. Then, they were observed under a high-power mirror, and the number was counted. It was found that the optimized Segnet network algorithm increased the IoU by 7.3%, the precision by 8.2%, and the recall rate by 11.1%. The diagnostic coincidence rates of conventional ultrasound and contrast-enhanced ultrasound examinations for gallbladder cancer were 78.13% (25/32) and 87.5% (25/32), respectively. The positive expression rate of P16 in gallbladder adenocarcinoma (47.06%) was significantly lower than that of acute cholecystitis with gallbladder stones (84.38%) and gallbladder polyps (67.16%) ( P < 0.05 ). The positive expression rate of P16 in patients with stage III and stage IV (33.33% and 40%) was significantly lower than that in patients with stages I and II (87.5% and 80%) ( P < 0.05 ). The positive expression rate of P16 in high differentiation (86.67%) was significantly higher than that of moderate differentiation (40%) and poor differentiation (28.57%) ( P < 0.05 ). In short, contrast-enhanced ultrasound can effectively improve the diagnostic coincidence rate of gallbladder cancer, and the expression of P16 in gallbladder cancer is closely related to tumor staging and differentiation.

Immunophenotyping of an Unusual Mixed-Type Extraskeletal Osteosarcoma in a Dog

A 6-year-old female Maltese dog presented with a cervical mass without pain. The tumor was surrounded by a thick fibrous tissue and consisted of an osteoid matrix with osteoblasts and two distinct areas: a mesenchymal cell-rich lesion with numerous multinucleated giant cells and a chondroid matrix-rich lesion. The tumor cells exhibited heterogeneous protein expression, including a positive expression of vimentin, cytokeratin, RANKL, CRLR, SOX9, and collagen 2, and was diagnosed as extraskeletal osteosarcoma. Despite its malignancy, the dog showed no sign of recurrence or metastasis three months after the resection. Further analysis of the tumor cells revealed a high expression of proliferation- and metastasis-related biomarkers in the absence of angiogenesis-related biomarkers, suggesting that the lack of angiogenesis and the elevated tumor-associated fibrosis resulted in a hypoxic tumor microenvironment and prevented metastasis.

Aldehyde dehydrogenase 3B2 promotes the proliferation and invasion of cholangiocarcinoma by increasing Integrin Beta 1 expression

Aldehyde dehydrogenases (ALDHs) play an essential role in regulating malignant tumor progression; however, their role in cholangiocarcinoma (CCA) has not been elucidated. We analyzed the expression of ALDHs in 8 paired tumor and peritumor perihilar cholangiocarcinoma (pCCA) tissues and found that ALDH3B1 and ALDH3B2 were upregulated in tumor tissues. Further survival analysis in intrahepatic cholangiocarcinoma (iCCA, n = 27), pCCA (n = 87) and distal cholangiocarcinoma (dCCA, n = 80) cohorts have revealed that ALDH3B2 was a prognostic factor of CCA and was an independent prognostic factor of iCCA and pCCA. ALDH3B2 expression was associated with serum CEA in iCCA and dCCA, associated with tumor T stage, M stage, neural invasion and serum CA19-9 in pCCA. In two cholangiocarcinoma cell lines, overexpression of ALDH3B2 promoted cell proliferation and clone formation by promoting the G1/S phase transition. Knockdown of ALDH3B2 inhibited cell migration, invasion, and EMT in vitro, and restrained tumor metastasis in vivo. Patients with high expression of ALDH3B2 also have high expression of ITGB1 in iCCA, pCCA, and dCCA at both mRNA and protein levels. Knockdown of ALDH3B2 downregulated the expression of ITGB1 and inhibited the phosphorylation level of c-Jun, p38, and ERK. Meanwhile, knockdown of ITGB1 inhibited the promoting effect of ALDH3B2 overexpression on cell proliferation, migration, and invasion. ITGB1 is also a prognostic factor of iCCA, pCCA, and dCCA and double-positive expression of ITGB1 and ALDH3B2 exhibits better performance in predicting patient prognosis. In conclusion, ALDH3B2 promotes tumor proliferation and metastasis in CCA by regulating the expression of ITGB1 and upregulating its downstream signaling pathway. The double-positive expression of ITGB1 and ALDH3B2 serves as a better prognostic biomarker of CCA.

Case Report: Response With Immunotherapy in a Patient With Mixed Neuroendocrine Non-Neuroendocrine Neoplasms of the Gallbladder

BackgroundPrimary neuroendocrine tumors of the gallbladder (GB-NETs) are rare, accounting for 2% of all gallbladder cancers. Among GB-NETs, mixed neuroendocrine non-neuroendocrine neoplasms of the gallbladder (GB-MiNENs) are sporadic.Case PresentationA 56-year-old woman admitted to our hospital due to right upper abdominal pain of 3 days duration. She underwent positron emission tomography/computed tomography, which showed multiple metastatic tumors and was unsuitable for operation. Initially, the patient was diagnosed with gallbladder adenocarcinoma. She underwent PD-1 inhibitor or combined with chemotherapy considering the PD-L1 high positive score. In the latter, the patient has the opportunity of surgery, and the new diagnosis was MiNENs. She achieved long-term disease control and has been alive from the first diagnosis.ConclusionThis case might support the strategy that PD-1 inhibitor could provide a feasible treatment option for MiNENs of gallbladder patients with the positive expression of PD-L1 in the future.

An Immunohistochemical Evaluation of Tumor-Associated Glycans and Mucins as Targets for Molecular Imaging of Pancreatic Ductal Adenocarcinoma

Targeted molecular imaging may overcome current challenges in the preoperative and intraoperative delineation of pancreatic ductal adenocarcinoma (PDAC). Tumor-associated glycans Lea/c/x, sdi-Lea, sLea, sLex, sTn as well as mucin-1 (MUC1) and mucin-5AC (MU5AC) have gained significant interest as targets for PDAC imaging. To evaluate their PDAC molecular imaging potential, biomarker expression was determined using immunohistochemistry on PDAC, (surrounding) chronic pancreatitis (CP), healthy pancreatic, duodenum, positive (LN+) and negative lymph node (LN−) tissues, and quantified using a semi-automated digital image analysis workflow. Positive expression on PDAC tissues was found on 83% for Lea/c/x, 94% for sdi-Lea, 98% for sLea, 90% for sLex, 88% for sTn, 96% for MUC1 and 67% for MUC5AC, where all were not affected by the application of neoadjuvant therapy. Compared to PDAC, all biomarkers were significantly lower expressed on CP, healthy pancreatic and duodenal tissues, except for sTn and MUC1, which showed a strong expression on duodenum (sTn tumor:duodenum ratio: 0.6, p < 0.0001) and healthy pancreatic tissues (MUC1 tumor:pancreas ratio: 1.0, p > 0.9999), respectively. All biomarkers are suitable targets for correct identification of LN+, as well as the distinction of LN+ from LN− tissues. To conclude, this study paves the way for the development and evaluation of Lea/c/x-, sdi-Lea-, sLea-, sLex- and MUC5AC-specific tracers for molecular imaging of PDAC imaging and their subsequent introduction into the clinic.

Correlation of the AKT/mTOR signaling pathway with the clinicopathological features and prognosis of nasopharyngeal carcinoma

The primary aim of this study was to examine the correlation of the AKT/mTOR signaling pathway with the clinicopathological features and prognostic significance in nasopharyngeal carcinoma (NPC). The study tissues were collected from 285 patients with NPC and normal mucosal tissues were obtained from 289 individuals with normal nasopharynxes. Immunohistochemical staining was used to detected the expression of the AKT, mTOR, and p70 ribosomal S6 kinase (P70S6K) proteins. Follow-up was performed for between 8 and 60 months. Spearman’s rank correlation analysis was performed to evaluate the correlation of the expression of the AKT, mTOR, and P70S6K proteins in NPC tissues. Kaplan-Meier curves were plotted to show the survival of patients with NPC. A Cox proportional hazards model was used to explore the independent risk factors for prognosis. The expression of the AKT, mTOR, and P70S6K proteins in NPC tissues was higher than that in healthy nasopharyngeal mucosal tissues, and was correlated with T-staging, N-staging, clinical stage, distant metastasis, and differentiation. The positive expression of the AKT, mTOR, and P70S6K proteins was higher in patients with stage III/IV NPC, low differentiation, and metastasis. The survival rates of patients with NPC with AKT-positive, mTOR-positive, and P70S6K-positive expression were considerably lower than those without the expression of these proteins. Distant metastasis and the overexpression of the AKT, mTOR, and P70S6 proteins were independent risk factors for the prognosis of patients with NPC. The results obtained from this study indicated an association between the AKT/mTOR signaling pathway and the progression of NPC. The upregulation of the AKT/mTOR pathway in patients with NPC is a predictor of poor prognosis.