Objective: To investigate the clinical and pathologic features, diagnosis and differential diagnosis of congenital hemangioma (CH). Methods: Forty cases of CH were diagnosed from January 2017 to December 2020 in Henan Provincial People's Hospital. The clinical and pathological and immunohistochemical data were analyzed, with review of literature. Results: There were 24 male and 16 female patients. The lesions were located in the head, neck (11 cases), limbs (14 cases), and trunk (15 cases). The clinical manifestations were congenital painless plaques or masses, the larger ones protruded on the skin surface, mostly dusky purple or bright red, with surrounding white halos. Under low magnification, the tumor was lobular and well demarcated, composed of neo-microvascular lumen of different sizes. The vascular endothelial cells were cuboidal or hobnail in appearance, forming stellar drainage vessels within the lobules. Extra-medullary hematopoiesis was seen in one case of rapidly involuting CH; there were different number of tortuous and dilated vascular lumen between the lobular structures, and some non-involuting CH cases were vascular malformations, which were devoid of lobulated structures. Immunohistochemistry showed that endothelial cells were strongly positive for CD31, CD34 and ERG, while D2-40 and GLUT-1 were negative. Conclusions: CH is a benign congenital vascular tumor with characteristic lobulated growth and abnormal blood vessels in the stroma. Pathological diagnosis often needs to be differentiated from infantile hemangioma, pyogenic granuloma, kaposiform hemangioendothelioma and vascular malformation.
Endothelial Cells/pathology* ; Female ; Hemangioendothelioma/pathology* ; Hemangioma/pathology* ; Humans ; Kasabach-Merritt Syndrome/pathology* ; Male ; Sarcoma, Kaposi/pathology* ; Skin Neoplasms/pathology*

Background: Porcine circovirus type 2 (PCV2) is an important infectious pathogen implicated in porcine circovirus-associated diseases (PCVAD), which has caused significant economic losses in the pig industry worldwide. Objectives: A suitable viral vector-mediated gene transfer platform for the expression of the capsid protein (Cap) is an attractive strategy. Methods: In the present study, a recombinant adeno-associated virus 8 (rAAV8) vector was constructed to encode Cap (Cap-rAAV) in vitro and in vitro after gene transfer. Results: The obtained results showed that Cap could be expressed in HEK293T cells and BABL/c mice. The results of lymphocytes proliferative, as well as immunoglobulin G (IgG) 2a and interferon-γ showed strong cellular immune responses induced by Cap-rAAV. The enzyme-linked immunosorbent assay titers obtained and the IgG1 and interleukin-4 levels showed that humoral immune responses were also induced by Cap-rAAV. Altogether, these results demonstrated that the rAAV8 vaccine Cap-rAAV can induce strong cellular and humoral immune responses, indicating a potential rAAV8 vaccine against PCV2. Conclusions The injection of rAAV8 encoding PCV2 Cap genes into muscle tissue can ensure long-term, continuous, and systemic expression.

The transition from spermatogonia to spermatocytes and the initiation of meiosis are key steps in spermatogenesis and are precisely regulated by a plethora of proteins. However, the underlying molecular mechanism remains largely unknown. Here, we report that Src homology domain tyrosine phosphatase 2 (Shp2; encoded by the protein tyrosine phosphatase, nonreceptor type 11 [Ptpn11] gene) is abundant in spermatogonia but markedly decreases in meiotic spermatocytes. Conditional knockout of Shp2 in spermatogonia in mice using stimulated by retinoic acid gene 8 (Stra8)-cre enhanced spermatogonial differentiation and disturbed the meiotic process. Depletion of Shp2 in spermatogonia caused many meiotic spermatocytes to die; moreover, the surviving spermatocytes reached the leptotene stage early at postnatal day 9 (PN9) and the pachytene stage at PN11-13. In preleptotene spermatocytes, Shp2 deletion disrupted the expression of meiotic genes, such as disrupted meiotic cDNA 1 (Dmc1), DNA repair recombinase rad51 (Rad51), and structural maintenance of chromosome 3 (Smc3), and these deficiencies interrupted spermatocyte meiosis. In GC-1 cells cultured in vitro, Shp2 knockdown suppressed the retinoic acid (RA)-induced phosphorylation of extracellular-regulated protein kinase (Erk) and protein kinase B (Akt/PKB) and the expression of target genes such as synaptonemal complex protein 3 (Sycp3) and Dmc1. Together, these data suggest that Shp2 plays a crucial role in spermatogenesis by governing the transition from spermatogonia to spermatocytes and by mediating meiotic progression through regulating gene transcription, thus providing a potential treatment target for male infertility.
Animals ; Cell Cycle Proteins/genetics* ; Cell Line ; Cell Survival ; Chondroitin Sulfate Proteoglycans/genetics* ; Chromosomal Proteins, Non-Histone/genetics* ; Gene Expression Regulation ; Gene Knockdown Techniques ; Infertility, Male ; Male ; Meiosis/genetics* ; Mice ; Mice, Knockout ; Mice, Transgenic ; Phosphate-Binding Proteins/genetics* ; Protein Tyrosine Phosphatase, Non-Receptor Type 11/genetics* ; Rad51 Recombinase/genetics* ; Real-Time Polymerase Chain Reaction ; Spermatocytes/metabolism* ; Spermatogenesis/genetics* ; Spermatogonia/metabolism*

Objective To investigate the clinicopathological characteristics and the expression of related molecular markers of heterotopic pancreas for better understanding and avoiding overtreatment of this disease.Methods From 24th March 2009 to 10th November 2016,92 patients with heterotopic pancreas in upper digestive tract diagnosed after endoscopic submucosal dissection(ESD),were collected. Tissues were sectioned and pathologically classified by Heinrich classification.The expressions of seven different molecular markers including cytokeratin(CK)19,insulin,trypsin,Ki-67,p53,CD133 and CD56 were detected by immunohistochemistry staining. Clinical features, pathological features and immunohistochemical results were retrospectively analyzed.Analysis of variance and Kruskal-Wallis test were used.Results According to Heinrich classification,the percentages of type Ⅰ,Ⅱ,and Ⅲ of heterotopic pancreas were 27.2%(25/92),63.0%(58/92)and 9.8%(9/92),respectively.There was no type Ⅳ patients.The heterotopic pancreas mainly located in stomach with proportion being about 91.3%(84/92)of the total heterotopic pancreas.CK19(the marker of pancreas ducts),insulin(as marker of endocrine function)and trypsin(as the marker of exocrine function)were all expressed in heterotopic pancreas.The positive rate of CD56,a pancreatic neuroendocrine marker,was 66.3%(61/92).The umbilicus like depression was the typical endoscopic appearance of heterotopic pancreas,which only found in 29 patients(31.5%).The average rate of Ki-67,cell proliferation index,was(2.08 ± 1.41)%.The expression of mutant p53 was negative in all 92 cases of heterotopic pancreas.The average staining area of CD133,a marker of pancreatic cancer stem cell,was(2.53 ± 2.43)%.The average follow-up period of 92 patients was(43.6 ± 27.5)months.No relapse and malignant change were found and all patients survived after ESD.Conclusions Heterotopic pancreas has normal pancreatic construction and function.The cell proliferation index is low in heterotopic pancreas and no mutant p53 expression is found.The expression of CD133 is also low.Heterotopic pancreas is a congenital benign disease which requires a long-term follow-up rather than overtreatment.

BACKGROUNDMorphological changes of the vasculature system in patients with myopia have been observed by Doppler ultrasound and fundus fluorescein angiography (FFA); however, these studies have limitations. Doppler ultrasound provides low-resolution images which are mainly obtained from visualized large vessels, and FFA is an invasive examination. Optic coherence tomography (OCT) angiography is a noninvasive, high-resolution measurement for vascular density. The purpose of this study was to investigate the change of vascular density in myopic eyes using OCT angiography.

METHODSThis cross-sectional study includes a total of 91 eyes from 47 participants including control, moderate, and high myopia that were evaluated by OCT angiography. Patients with myopia were recruited from the Refractive Department, Shenzhen Aier Eye Hospital, from August 5, 2015 to April 1, 2016. Emmetropic eyes were from healthy volunteers. The vascular density at macula and optic disc regions, ganglion cell complex (GCC) thickness, and retinal nerve fiber layer (RNFL) thickness were measured. Their relationships with axial length (AL) and refractive error were analyzed. One-way analysis of variance (ANOVA), Pearson's correlation, and generalized estimating equation were used for statistical analysis.

RESULTSBoth superficial and deep macular vascular density were highest in control (25.64% ± 3.76% and 37.12% ± 3.66%, respectively), then in moderate myopia (21.15% ± 5.33% and 35.35% ± 5.50%, respectively), and lowest in high myopia group (19.64% ± 3.87% and 32.81% ± 6.29%, respectively) (F = 13.74 and 4.57, respectively; both P < 0.001). Both superficial (β = -0.850 and 0.460, respectively) and deep (β = -0.766 and 0.396, respectively) macular vascular density were associated with AL and spherical equivalent (all P < 0.001). Superficial macular vascular density was associated with GCC thickness (β = 0.244, P = 0.040), independent of spherical equivalent. The vascular density in optic disc region had no difference among the three groups, and it was not associated with AL, spherical equivalent, or RNFL thickness.

CONCLUSIONOur results suggested that with the increase of myopia, the vascular density decreased in macular region, but not in optic disc region.

Adult ; Cross-Sectional Studies ; Eye ; blood supply ; Female ; Fluorescein Angiography ; Humans ; Macula Lutea ; pathology ; physiopathology ; Male ; Middle Aged ; Myopia ; pathology ; physiopathology ; Optic Disk ; pathology ; physiopathology ; Prospective Studies ; Retina ; pathology ; physiopathology ; Retinal Ganglion Cells ; pathology ; Tomography, Optical Coherence ; Young Adult

Objective To evaluate the potential malignancy, prognosis and risk factors for intraductal papillary mucinous neoplasm(IPMN), which were classified into different risk levels based on Fukuoka guideline. Methods A retrospective analysis of patients with IPMN diagnosed at Nanjing Drum Tower Hospital from 2009 to 2016 was conducted. Clinical characteristics,treatment and prognosis of IPMNs were analyzed. Results A total of 94 IPMN patients were included and divided into 3 groups according to Fukuoka guideline,46 patients in high-risk(HR)group,30 in group of worrisome features(WF), and 18 in low-risk(LR)group. For patients undergoing surgery treatment, there were 5 cases(19.2%,5/26)in HR group and 2 cases(12.5%,2/16)in WF group whose postoperative pathological findings were malignant (P=0.690). The 5-year survival rates after operations were 73.9% and 77.0% in HR and WF group, respectively(P=0.830). For patients without surgery treatment, in a 5-year follow-up, there were 6 cases (33.3%,6/18),2 cases(16.7%,2/12)and 0(0.0%,0/18)progressing into pancreatic cancers in HR, WF and LR groups,respectively(P<0.05). In addition,among the three groups,the 5-year survival rates were 49.5%,85.7% and 100.0%(P=0.025). Jaundice was significantly related to prognosis(P<0.01) and the hazard ratio was 8.883(95%CI:2.953-26.721). Conclusion Jaundice is a predictive risk factor for survival of IPMN. As for the treatment to IPMN, patients in HR group should receive surgery treatment while those in LR group can be followed up. For patients in WF group,the treatment should be customized, with evaluation of predictive risk factors,and operations can be performed when needed.

The role of (pro)rennin receptor (PRR) in cardiomyocytes of a heart failure (HF) rat model was studied. Spontaneously hypertensive rats (SHR) with HF (SHR-HF) or not were identified by two-dimensional (2-D) ultrasound. Age-matched Wistar Kyoto normotensive (WKY) rats were used as controls. PRR short hair RNA (sh-RNA) was injected into the heart of SHR-HF. Simultaneously SHR and controls received the same shRNA injection into the heart. Scramble shRNA was injected into the heart as controls. The expression of PRR mRNA and protein in cardiomyocytes was detected by using real-time PCR and Western blotting respectively. The heart function was evaluated by 2-D ultrasound, including eject fraction (EF%), fractional shortening (FS%), left ventricle thickness (LV), and inter-ventricular septal thickness (IVS). The number of apoptotic cardiomyocytes was counted by using flow cytometry. The results showed that the mRNA and protein expression levels of PRR were significantly higher in cardiomyocytes of SHR-HF group than in those of SHR group or control group. The apoptosis of myocytes in SHR-HF group was increased as compared with SHR group or control group. After knock-down of PRR with shRNA in SHR-HF group, the apoptosis of myocytes was reduced, resulting in the improved heart function. It was suggested that down-regulation of PRR might protect the heart from development of HF in SHR-HF by inhibiting the apoptosis of cardiomyocytes.
Animals ; Apoptosis ; genetics ; physiology ; Blotting, Western ; Chymosin ; metabolism ; Disease Models, Animal ; Enzyme Precursors ; metabolism ; Gene Expression ; Heart Failure ; genetics ; metabolism ; physiopathology ; Male ; Myocardium ; metabolism ; pathology ; Myocytes, Cardiac ; metabolism ; RNA Interference ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Receptors, Cell Surface ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

To study the genetic characteristics of 123 type II non-wild polioviruses isolated from acute flaccid paralysis (AFP) cases in mainland China in 2010, provide the scientific basis for maintaining the "polio-free" status, and the switching use of polio vaccine for China. VP1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the PCR products were then sequenced. The sequence results were analyzed with Sequencher 4.8, BioEdit 7.0.9 and MEGA 5.0. Of 65 strains, nt2909 was found to be a mutation hotspot, and also a neurovirulence determinant in VP1 region. During 2010, two vaccine-derived polioviruses (VDPVs) were isolated from Yunnan province, China and no wild poliovirus (WPV) was isolated. The epidemiological studies and laboratory results of the two VDPVs showed that they were newly discovered VDPVs because of the genetic difference from other VDPVs strains isolated in the world, implying the sensitive poliovirus surveillance network could timely detect the transmission of VDPVs and the importation of WPV.
Adolescent ; Adult ; Child ; Child, Preschool ; China ; Female ; Genotype ; Humans ; Male ; Phylogeny ; Poliomyelitis ; virology ; Poliovirus ; classification ; genetics ; isolation & purification ; Viral Proteins ; genetics ; Young Adult

OBJECTIVETo investigate the anti-tumor activity of combined gemcitabine with dihydroartemisinin, and the mechanism of the anti-tumor effect of gemcitabine enhanced by dihydroartemisinin on pancreatic cancer.

METHODSFor cultured cells, cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis and confocal laser scanning microscope stained with Annexin V-FITC/PI. The nuclear extract for determining NF-kappaB DNA-binding activity was analyzed by EMSA, while nuclear P65 and its downstream gene expression was determined by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to agents. TUNEL assay was used to assess tumor cell apoptosis in tumor tissue.

RESULTSAfter combination of gemcitabine and dihydroartemisinin treatment, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and Panc-1 reached up to (81.1 +/- 3.9)% and (76.5 +/- 3.3)%, and the apoptosis rates were up to (53.6 +/- 3.8)% and (48.3 +/- 4.3)%, the differences were significantly (P < 0.01) compared with gemcitabine [(24.8 +/- 2.9)% and (21.8 +/- 3.5)%]. All the treatment groups inhibited the growth of pancreatic xenograft tumors in nude mice. The tumor volume and apoptosis index were (262 +/- 37) mm(3) and (50 +/- 4)% respectively in the combined treatment, compared to those of [(384 +/- 56) mm(3) and (25 +/- 3)%] in gemcitabine, the differences were significantly (P < 0.05). EMSA showed that gemcitabine alone obviously enhanced its DNA-binding activity compared to control. However, dihydroartemisinin significantly reduced its DNA-binding activity, so that abrogated the inducing effect of gemcitabine on NF-kappaB activation. Western blot assay indicated that dihydroartemisinin downregulated expression of nuclear P65, and combined treatment not only downregulated the expression of Cyclin D1, Bcl-xL and Bcl-2 while upregulated Bax, thus reduced the Bcl-2/Bax ratio, but also increased the caspase-3 activation, all of which increased apoptosis in both BxPC-3 and Panc-1 cells.

CONCLUSIONDihydroartemisinin significantly abrogated the inducing effect of gemcitabine on NF-kappaB activation and downregulated the expression of NF-kappaB targeted gene products, which may be one possible mechanism by which dihydroartemisinin augments the anti-tumor effect of gemcitabine on pancreatic cancer.

Animals ; Antimetabolites, Antineoplastic ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Apoptosis ; drug effects ; Artemisinins ; therapeutic use ; Cell Line, Tumor ; Deoxycytidine ; analogs & derivatives ; therapeutic use ; Humans ; Male ; Mice ; Mice, Nude ; NF-kappa B ; metabolism ; Pancreatic Neoplasms ; drug therapy ; metabolism ; pathology ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate the effect and mechanism of NF-kappaB P65 gene silencing by small interference RNA on the apoptosis of human pancreatic cancer cells induced by gemcitabine in vitro and in vivo.

METHODSHuman pancreatic cancer cells (BxPC-3 and PANC-1) were cultured and respectively divided into five groups: blank control group, negative control siRNA group, gemcitabine group, NF-kappaB P65 siRNA group and gemcitabine + P65 siRNA group. The ability of cell proliferation was analyzed by MTT; the expression of NF-kappaB P65 and the apoptosis related proteins were examined by Western blot assay; the apoptosis was evaluated by the flow cytometry and laser scanning confocal microscopy analysis stained with Annexin V-FITC/PI; the DNA binding activity of NF-kappaB was examined by electrophoretic mobility shift assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors. The tumor volume was monitored and TUNEL assay was used to assess the apoptosis index in tumor tissue after treatment.

RESULTSAt 72 h after transfection, the combination with gemcitabine and p65 siRNA significantly decreased the cell viability index (P < 0.05), and down-regulated the expression of Bcl-2 and procaspase-3 and up-regulated the expression of Bax compared with other groups. The combined treatment significantly increased the rate of apoptosis compared with other groups (P < 0.05). EMSA assay indicated that the DNA binding activity of NF-kappaB significantly decreased in NF-kappaB P65 siRNA group and gemcitabine+P65 siRNA group compared with Control group. The combined therapy inhibited the growth of pancreatic xenograft tumors by apoptosis induction in nude mice (P < 0.01).

CONCLUSIONSThe effect of gemcitabine inducing cell apoptosis may be potentiated through inhibiting the DNA binding activity of NF-kappaB and regulating the expression of apoptosis related proteins by NF-kappaB P65 siRNA, which can activate the mitochondria apoptosis pathway in pancreatic cancer in vitro and in vivo.

Animals ; Apoptosis ; drug effects ; genetics ; Cell Line, Tumor ; Deoxycytidine ; analogs & derivatives ; pharmacology ; Humans ; Mice ; Mice, Inbred BALB C ; Pancreatic Neoplasms ; drug therapy ; metabolism ; pathology ; RNA, Small Interfering ; genetics ; Transcription Factor RelA ; genetics ; metabolism ; Transfection ; Xenograft Model Antitumor Assays