Synergistic effects of methotrexate and piperine on human osteosarcoma cell lines – An in-vitro study

Background: Osteosarcoma is a malignant cancer that effect bone and metastasizing to many vital organs such as lungs. There are many available drugs to treat the disease including tamoxifen, methotrexate (MTX), and cisplatin which have their own side effects and hurdles to become drugs of choice for the disease. On the other hand, introduction of herbal drugs as chemotherapeutic agents opened up new arena to potentiate the existing treatment by exhibiting synergy. Piperine (PPN) is widely used drug as anti-cancer agent as well as it has anti-inflammatory, analgesic properties, and also used in the treatment of abdominal pains, tuberculosis, arthritis, and respiratory illness. Aims and Objective: Thus, this study was designed to investigate the synergistic inhibitory potential of PPN and MTX on the MG63 osteosarcoma cell lines in vitro. Materials and Methods: The cell lines were cultured on DMEM medium and investigated for cytotoxicity of the drugs using MTT assay at 540 nm in UV. Three groups of cell lines administered with PPN, MTX, and PPN+MTX (1:1) in various concentrations and IC50 values were calculated based on the % cell viability graphs. Results: Results showed that the IC50 of PPN was 38.65, MTX was 123.98, and PPN+MTX was 15.13 proving the significant synergistic cytotoxic effect of PPN and MTX in inhibiting the proliferation of MG63 cell lines. Conclusion: Further research needs to be conducted in this field to elucidate the synergistic pathways in which PPN has shown a better anti-osteosarcoma effect when combined with MTX.

Expression profile in Leishmania major exposed to Staphylococcus aureus and group A beta-hemolytic Streptococcus

Background & objectives: The interaction of Leishmania spp. with microbiota inside the midgut vector has significant output in pathogenesis. This study aimed to identify the profile of Leishmania major gene expression of LACK, gp63, and hsp70 after exposure to Staphylococcus aureus and group A beta-hemolytic Streptococci (GABHS). Methods: Leishmania major (MRHO/IR/75/ER) promastigotes were exposed with S. aureus, with GABHS, and with both GABHS and S. aureus at 25°C for 72 h. The gene expression analysis of Lmgp63, Lmhsp70, and LmLACK was assessed using SYBR Green real-time PCR by ??Ct. All experiments were repeated in triplicate. Statistical analysis was done using two-way ANOVA. A P-value less than 0.05 was considered significant. Results: Lmgp63 was expressed in the group exposed to GABHS with 1.75-fold lower than the control group (p=0.000). The LmLACK had expression in both groups exposed with GABHS and GABHS with S. aureus with 2.8 and 1.33-fold more than the control group, respectively (p=0.000). The Lmhsp70 gene expression was reported in the group exposed with GABHS with relative quantification of 5.7-fold more than the control group. Interpretation & conclusion: This study showed that the important genes encoding LACK, gp63, and hsp70 changed their expression after exposure to the S. aureus and GABHS.

Butorphanol affects the proliferation, migration and invasion of osteosarcoma MG-63 cells via Hippo/YAP signaling pathway / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨布托啡诺（BPH）对骨肉瘤（OS）细胞增殖、迁移和侵袭的影响及其相关的作用机制。方法：将MG-63细胞分为对照组、YAP抑制剂组（维替泊芬组）和BPH低、中、高浓度组，MTT法、克隆形成实验、FCM术、划痕愈合实验、Transwell实验、qPCR法、WB法和移植瘤实验分别检测处理后各组细胞的增殖活性、克隆形成数、细胞凋亡率、划痕愈合率，以及上皮钙黏蛋白（E-cadherin）、神经钙黏蛋白（N-cadherin）、波形蛋白（vimentin）mRNA的表达和YAP、TAZ蛋白的表达，同时观察BPH和维替泊芬对移植瘤生长的影响。结果：与对照组相比，维替泊芬组和BPH低、中、高浓度组细胞增殖活性、克隆数、划痕愈合率、侵袭细胞数，以及N-cadherin和vimentin mRNA水平、YAP和TAZ蛋白表达及移植瘤体积均显著降低（均P<0.05），细胞凋亡率、E-cadherin mRNA水平及对移植瘤的抑瘤率均升高（均P<0.05），且BPH高浓度组与维替泊芬组之间各项指标均无明显差异（均P>0.05）。结论：BPH可能通过抑制Hippo/YAP信号通路来抑制OS细胞MG-63增殖、迁移和侵袭。

Protease inhibitors as a potential agent against visceral Leishmaniasis: A review to inspire future study

Abstract Leishmaniasis is transmitted by sandfly which carries the intracellular protozoa in their midgut. Among visceral, cutaneous and mucocutaneous leishmaniasis, visceral type that is caused by Leishmania donovani is the most lethal one. Findings of leishmanial structure and species took place in 19th century and was initiated by Donovan. Leishmaniasis is still a major concern of health issues in many endemic countries in Asia, Africa, the Americas, and the Mediterranean region. Worldwide1.5-2 million new cases of cutaneous leishmaniasis and 500,000 cases of visceral leishmaniasis are reported each year. Leishmaniasis is endemic in nearly 90 countries worldwide and close to 12 million new cases of leishmaniasis are reported worldwide annually. Studies on antileishmanial drug development is of major concern as leishmaniasis are the second largest parasitic killer in the world and the available drugs are either toxic or costly. The major surface GP63 protease, also known as Zinc- metalloproteases present on the surface of leishmanial promastigotes, can be targeted for drug development. Protease inhibitors targeting such surface proteases show promising results. Different protease inhibitors have been isolated from marine actinobacteria against many infectious diseases. Metabolites produced by these actinobacteria may have greater importance for the discovery and development of new antileishmanial drugs. Hence, this review discusses the background, current situation, treatment, and protease inhibitors from marine actinobacteria for drug development against GP63 molecules.

Polyunsaturated fatty acids alter the formation of lipid droplets and eicosanoid production in Leishmania promastigotes

BACKGROUND The knowledge about eicosanoid metabolism and lipid droplet (LD) formation in the Leishmania is very limited and new approaches are needed to identify which bioactive molecules are produced of them. OBJECTIVES Herein, we compared LDs and eicosanoids biogenesis in distinct Leishmania species which are etiologic agents of different clinical forms of leishmaniasis. METHODS For this, promastigotes of Leishmania amazonensis, L. braziliensis and L. infantum were stimulated with polyunsaturated fatty acids (PUFA) and LD and eicosanoid production was evaluated. We also compared mutations in structural models of human-like cyclooxygenase-2 (GP63) and prostaglandin F synthase (PGFS) proteins, as well as the levels of these enzymes in parasite cell extracts. FINDINGS PUFAs modulate the LD formation in L. braziliensis and L. infantum. Leishmania spp with equivalent tissue tropism had same protein mutations in GP63 and PGFS. No differences in GP63 production were observed among Leishmania spp, however PGFS production increased during the parasite differentiation. Stimulation with arachidonic acid resulted in elevated production of hydroxyeicosatetraenoic acids compared to prostaglandins. MAIN CONCLUSIONS Our data suggest LD formation and eicosanoid production are distinctly modulated by PUFAS dependent of Leishmania species. In addition, eicosanoid-enzyme mutations are more similar between Leishmania species with same host tropism.

Association of human papillomavirus (HPV), p16, p53 and p63 expression with non-bilharzia-associated squamous cell carcinoma of the bladder and algorithm construction for histopathological grading prediction

ABSTRACT Objective To investigate the expression of human papillomavirus (HPV), p16, p53, and p63 in non-schistosomiasis-related squamous cell carcinoma of the bladder and to develop an accurate and automated tool to predict histological classification based on clinicopathological features. Methods Twenty-eight patients with primary bladder pure squamous cell carcinoma who underwent cystectomy or transurethral resection of bladder tumor (TURBT) for bladder cancer between January 2011 and July 2017 were evaluated. Clinical data and follow-up information were obtained from medical records. Formalin-fixed, paraffin-embedded surgical specimens were used for immunohistochemical staining for p16, p53, and p63. Human papillomavirus detection was evaluated by PCR. Statistical analysis was performed, and statistical significance was set at p<0.05. Finally, decision trees were built to classify patients' prognostic features. Leave-one-out cross-validation was used to test the generalizability of the model. Results Neither direct HPV detection nor its indirect marker (p16 protein) was identified in most cases. The absence of p16 was correlated with less aggressive histological grading (p=0.040). The positive p16 staining detection found only in pT1 and pT2 cases in our sample suggests a possible role for this tumor suppressor protein in the initial stages of bladder squamous cell carcinoma. The decision trees constructed described the relationship between clinical features, such as hematuria/dysuria, the level of tumor invasion, HPV status, lymphovascular invasion, gender, age, compromised lymph nodes, and tumor degree differentiation, with high classification accuracy. Conclusion The algorithm classifier approach established decision pathways for semi-automatic tumor histological classification, laying the foundation for tailored semi-automated decision support systems for pathologists.

Mechanosensitive Ion Channel TMEM63A Gangs Up with Local Macrophages to Modulate Chronic Post-amputation Pain / 神经科学通报·英文版

Post-amputation pain causes great suffering to amputees, but still no effective drugs are available due to its elusive mechanisms. Our previous clinical studies found that surgical removal or radiofrequency treatment of the neuroma at the axotomized nerve stump effectively relieves the phantom pain afflicting patients after amputation. This indicated an essential role of the residual nerve stump in the formation of chronic post-amputation pain (CPAP). However, the molecular mechanism by which the residual nerve stump or neuroma is involved and regulates CPAP is still a mystery. In this study, we found that nociceptors expressed the mechanosensitive ion channel TMEM63A and macrophages infiltrated into the dorsal root ganglion (DRG) neurons worked synergistically to promote CPAP. Histology and qRT-PCR showed that TMEM63A was mainly expressed in mechanical pain-producing non-peptidergic nociceptors in the DRG, and the expression of TMEM63A increased significantly both in the neuroma from amputated patients and the DRG in a mouse model of tibial nerve transfer (TNT). Behavioral tests showed that the mechanical, heat, and cold sensitivity were not affected in the Tmem63a-/- mice in the naïve state, suggesting the basal pain was not affected. In the inflammatory and post-amputation state, the mechanical allodynia but not the heat hyperalgesia or cold allodynia was significantly decreased in Tmem63a-/- mice. Further study showed that there was severe neuronal injury and macrophage infiltration in the DRG, tibial nerve, residual stump, and the neuroma-like structure of the TNT mouse model, Consistent with this, expression of the pro-inflammatory cytokines TNF-α, IL-6, and IL-1β all increased dramatically in the DRG. Interestingly, the deletion of Tmem63a significantly reduced the macrophage infiltration in the DRG but not in the tibial nerve stump. Furthermore, the ablation of macrophages significantly reduced both the expression of Tmem63a and the mechanical allodynia in the TNT mouse model, indicating an interaction between nociceptors and macrophages, and that these two factors gang up together to regulate the formation of CPAP. This provides a new insight into the mechanisms underlying CPAP and potential drug targets its treatment.

p63-positive prostate carcinoma—A very rare presentation

Prostate cancer being the world's leading cause of cancer and also the second most common cancer in men is posing challenges in its diagnosis. Immunohistochemistry with markers like high molecular weight cytokeratin, p63 aid in the diagnosis. The absence of p63 and high molecular weight cytokeratin and presence of p504s in the biopsies indicate malignant lesions. Yet, there is a loophole to this too. A rare case of p63-positive prostatic adenocarcinoma in an 87-year-old patient, with immunohistochemistry results showing overexpression of p63 in the nuclei of the malignant glands. This tumor shows high molecular weight cytokeratin negativity, and p504s positivity. Prognosis of this variant of the tumor is mostly favorable. Prompt treatment will halt the progression of this tumor and prevent paraplegia. Radical prostatectomy could be avoided by treatment modalities like androgen blockade and brachytherapy, as morbidity is very high with radical prostatectomy surgery.

LYMPHOEPITHELIOMA LIKE CARCINOMA OF URINARY BLADDER: A RARE CASE REPORT

Urinary bladder neoplasm is the 10 most common cancer in the world, predominantly affecting males. Nowadays incidence of urinary bladder carcinoma in female has increased. It is a uncommon type carcinoma of urinary bladder. It resembles to non-keratinizing carcinoma of nasopharynx. This study report a case of 65 years old male presented with painless gross hematuria for 3 weeks. Patient was a known case of Meckel's diverticulum. Ultrasonography and cystoscopy showed a mass on lateral wall of urinary bladder measuring 3x3 cm . A radical cystectomy of the urinary bladder tumor was done. He was diagnosed as case of lymphoepithelioma like carcinoma of urinary bladder along with benign prostate hyperplasia and Meckel's diverticulum was made with stage 2. According to the WHO classification criteria, Lymphoepithelioma like carcinoma is a subtype of undifferentiated carcinomas. Immunohistochemical stains are helpful to differentiate Lymphoepithelioma from primary lymphoma of the bladder. Differentiation of LELCB from urothelial carcinoma is important, since it has better prognosis compared to pure urothelial carcinoma. It is important to diagnose this neoplasm for therapeutic as well as prognostic purpose.

Biochemical Properties of Parasite Virulence Factor: Lesson Learned from Leishmania

Leishmania, a parasitic protozoan, a single-celled organism of the genus trypanosomes that are responsible for the disease leishmaniasis. Transmission occured by sandflies of the genus Phlebotomus in the Old World, and of the genus Lutzomyia in the New World. Globally, at least 93 sandfly species are proven or probable vectors. Their primary hosts are vertebrates; Leishmania commonly infects hyraxes, canids, rodents, and humans. Leishmaniasis encompasses diverse clinical syndromes, including cutaneous, mucosal, and potentially life-threatening visceral forms. Three widely known virulence factors belongs to the genus Leishmania include the active compound named proteophosphoglycan (PPG), GP63 metalloprotease and lipophosphoglycan (LPG). these substance established on the surface of the parasite. The aim of this review article is to make an insight of the biochemical characteristics of Leishmania spp virulence factors, the armamentarium that predispose their pathogenesis, its invasion and virulence to the mammalian host.

miR-185 inhibits the proliferation and migration of osteosarcoma MG63 cells via regulating CDC42 gene expression / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To analyze the expression of miR-185 and cell division cyclin 42 (CDC42) in osteosarcoma tissues and cells, and to preliminarily explore whether miR-185 affects the proliferation and migration of osteosarcoma MG63 cells by regulating CDC42. Methods: The cancer tissues and para-cancerous tissues of 28 patients with osteosarcoma that pathologically confirmed in the Fourth People's Hospital of Hengshui City from January 2020 to January 2021 were collected for this study. Immunohistochemistry was used to detect the expression of CDC42 in osteosarcoma tissues, and qPCR was used to detect the expression of miR-185 in osteosarcoma tissues. Dual-luciferase reporter gene experiment was applied to verify the targeting relationship between CDC42 and miR-185. According to different transfectants, MG63 cells were divided into miR-185 mimic group, miR-NC group, miR-185 inhibitor group, NC-inhibitor group, CDC42 group (transfected with CDC42 over-expression vector), and negative control (NC) group. The effects of miR-185 and CDC42 expression on the migration, proliferation and cell cycle of MG63 cells were detected by scratch healing assay, CCK-8 method and FCM, respectively. A nude mouse xenograft model was constructed by inoculating osteosarcoma MG63 cells. Immunohistochemistry, qPCR and WB methods were used to detect the effects of over-expression or knock-down of miR-185 on the expression of Ki67 and CDC42 in transplanted tumor tissues. Results: Compared with para-cancerous tissues, the expression of miR-185 in osteosarcoma tissues was significantly decreased, while the expression of CDC42 was significantly increased (all P<0.01). CDC42 was verified to be a target gene of miR-185. Compared with the control group, the migration and proliferation of MG63 cells in the miR-185 mimic group were inhibited (all P<0.01), while the migration and proliferation of MG63 cells in the CDC42 group were increased and the cell cycle was arrested in the S phase (all P<0.01). Compared with the miR-185 group, the migration and proliferation abilities of MG63 cells in the miR-185+CDC42 group were promoted, and the proportion of cells in S phase was increased (all P<0.01). Compared with the control group, the expression of Ki67 and CDC42 in the transplanted tumor tissues of miR-185 mimic group was significantly decreased (all P<0.01), while the opposite results were observed in miR-185 inhibitor group (all P<0.01). Conclusion: miR-185 is lowly expressed while CDC42 is highly expressed in osteosarcoma tissues. miR-185 can inhibit the proliferation and migration of osteosarcoma MG63 cells by negatively regulating the expression of CDC42.

A case of ADULT syndrome without finger/toe deformities caused by a TP63 mutation / 中华皮肤科杂志

Objective:To detect causative gene mutations in 1 patient with ADULT syndrome mainly presenting with ectodermal dysplasia.Methods:Clinical data were collected from a proband with ADULT syndrome, and genomic DNA was extracted from peripheral blood samples obtained from the proband and his parents. Exome sequencing was performed in the proband by using targeted panels for hereditary skin diseases to determine mutation sites, and then the candidate mutation sites were verified by Sanger sequencing in the family members.Results:The 22-year-old male patient presented with sparse and thin hair, scattered facial freckles, missing permanent teeth, cloudy corneas, palmoplantar erythema and keratosis, nail/toenail dystrophy, and nipple dysplasia. Genetic testing of the peripheral blood genomic DNA of the proband revealed a heterozygous mutation (c.1040G>T) in exon 8 of the TP63 gene, resulting in an amino acid change at position 347 (p.C347F) . The mutation was not detected in his father or mother with normal phenotypes, suggesting the cosegregation of the gene mutation with the disease phenotype in the family.Conclusion:The de novo heterozygous missense mutation in the TP63 gene may be the causative mutation in the proband, and combined with clinical manifestations, the proband was diagnosed with ADULT syndrome without finger/toe deformities.

TP63 gene mutation analysis in a case of ankyloblepharon-ectodermal defects-cleft lip/palate syndrome / 中华皮肤科杂志

Objective:To analyze pathogenic mutations in a child with ankyloblepharon-ectodermal defects-cleft lip/palate syndrome.Methods:Clinical data were collected from a patient with ankyloblepharon-ectodermal defects-cleft lip/palate syndrome, and DNA was extracted from peripheral blood samples from the patient and his parents. High-throughput sequencing was performed in the patient by using a gene panel targeting hereditary skin diseases, aiming to determine sites of disease-causing gene mutations. Then, Sanger sequencing was conducted to bidirectionally verify the mutations in the patient and his parents.Results:The male patient aged 3 years and 9 months, and presented with extensive erythema, scales, erosions as well as repeated infections and erosions of the scalp after birth. Reticulated hyper- and hypopigmented patches and scars left on the trunk and limbs after healing of erosions. Physical examination also showed sparse scalp hair, absence of most eyebrows and eyelashes, cleft palate, dysplastic teeth, dystrophic finger and toe nails, and deformed ears without ankyloblepharon. Genetic testing of the patient showed a novel heterozygous missense mutation c.1790T>A (p.Ile597Asn) in the TP63 gene, which had not been reported previously and was rated as pathogenic according to the American College of Medical Genetics and Genomics guidelines. This mutation was not identified in either of his parents.Conclusion:The novel heterozygous missense mutation c.1790T>A in the TP63 gene probably contributes to ankyloblepharon-ectodermal defects-cleft lip/palate syndrome in the patient, which expands genotypic and phenotypic spectrum of this disease.

Effect of Lactobacillus coryniformis FZU63 on the flavor quality of black tea beverage / 生物工程学报

The tea beverages will be endowed with distinct aroma and taste, as well as various biologically active compounds including probiotic factors, when fermented with lactic acid bacteria (LAB). However, at present, few studies on the dynamics of flavors in tea soup at different fermentation stages were conducted. In this study, the composition of monosaccharides, aromatic components, free amino acids, and organic acids were measured, when the black tea beverages were fermented with Lactobacillus coryniformis FZU63 which was isolated from Chinese traditional kimchi. The results indicated that monosaccharides including glucose, fructose, mannose and xylose in black tea beverages are the main carbon sources for fermentation. In addition, the abundance of aromatic compounds in black tea soup are increased significantly at different fermentation stages, which endow the fermented black tea soup with fruit aroma on the basis of flowery and nutty aroma. Moreover, some bitter amino acids are reduced, whereas the content of sweet and tasty amino acids is elevated. Furthermore, the levels of lactic acid, malic acid, citric acid and other organic acids are accumulated during the fermentation. Additionally, sensory evaluation displays that black tea beverage is acquired with comprehensive high-quality after being fermented for 48 h. This study provides a theoretical basis to steer and control the flavor formation and quality of the fermented tea beverages during LAB fermentation.

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus)

ABSTRACT: This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.

RESUMO: Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus) / Fenotipagem citométrica do sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus)

This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.(AU)

Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.(AU)

Effects of calcium sulfate on proliferation of osteoblast-like MG-63 cells and osteoprotegerin/receptor activator of NF-κB ligand/receptor activator of NF-κB system / 中华创伤骨科杂志

Objective:To investigate the effects of calcium sulfate on the proliferation of osteoblast-like MG-63 cells and the osteoprotegerin/receptor activator of NF-κB ligand/receptor activator of NF-κB (OPG/RANKL/RANK) system.Methods:The extract of calcium sulfate was prepared. The osteoblast-like MG-63 cells were cultured for 24 hours in the medium containing calcium sulfate (the calcium sulfate group) and in the normal medium without calcium sulfate (the blank group), respectively. The growth of osteoblast-like MG-63 cells was observed and their proliferation detected by CCK-8. The mRNA and protein expression levels of OPG/RANKL were detected.Results:The growth of osteoblast-like MG-63 cells was fine in both groups. The CCK-8 test showed that the absorbance value at 24 h was 0.997±0.008 for the calcium sulfate group, significantly higher than that for the blank group (0.640±0.003) ( P<0.001). Respectively, the mRNA expression levels of OPG were 2.834±0.176 and 1.005±0.102 and the mRNA expression levels of RANKL 0.355±0.035 and 1.002±0.068 for the calcium sulfate group and the blank control group, showing statistically significant differences ( P<0.001). The results of Western blot showed that compared with the blank control group, the protein expression of OPG in osteoblast-like MG-63 cells was promoted but the protein expression of RANKL inhibited in the calcium sulfate group. Conclusion:Calcium sulfate may have a positive effect on bone formation, because it can promote the proliferation and activity of osteoblast-like MG-63 cells and regulate the OPG/RANKL/RANK system.

Allelic Diversity of the Hemochromatosis Gene (HFE) in Malays, Chinese and Indians

@#Introduction: Hereditary hemochromatosis (HH) is an autosomal recessive disorder that causes accumulation of iron in circulating blood and organs. The disease is associated with H63D, S65C and C282Y variants of the haemochromatosis (HFE) gene and, if not treated can cause organ damage and may prove fatal. The main objectives of the present survey were to screen these genetic variants and establish risk profiles for developing HH in Malays, Chinese and Indians. Methods: A total of two hundred and twenty-two unrelated and healthy individuals together representing Malay, Chinese and Indian ethnicities in Malaysia were scored for the H63D, S65C and C282Y variants using a polymerase chain reaction-restriction fragment length polymorphism technique. Results: There are clear differences in H63D, S65C and C282Y allele and genotype frequency distributions between Malays, Chinese and Indians. In particular, H63D is more common in Chinese (5.19%) and Indians (7.29%), while S65C is more common in Malays (1.03%) and Chinese (1.04%). In addition, a susceptibility genotype for HH (the compound heterozygote for C282Y and H63D) was only detected in Indians (0.02%). Conclusion: Overall, our study is the first to provide data on the prevalence of H63D, S65C, and C282Y genetic variants and HH risk profiles for Malays, Chinese and Indians.

Histopathological and immunohistochemical characteristic for lung cancer among Mongolian patients / Шинэ санаа Шинэ нээлт

Background@#Lung cancer is the leading cause of cancer death in both men and women. In the past decade, histopathological classification of lung cancer has become increasingly important in selecting the appropriate treatment.@*Methods@#All cases of lung cancer diagnosed pathologically in the last 2.5-year period were retrospectively identified at National Cancer Center and performed by descriptive case series design.@*Results@#The average age of our participants was 62, of which 82.2% were men. A total of 214 cases of lung cancer were reported, including 142 (66.4%) squamous cell carcinoma (SCC), 34 (16%) adenocarcinoma (AC), 4 (1.7%) NSCLC, 29 (13.6%) small cell carcinoma (SmCC) and 5 (2.3%) cases of other/unspecified type. Based on only morphology, initially 32 (17.7%) cases of NSCLC were diagnosed, whereas after immunohistochemistry (TTF-1 or NAPA positive/p63 negative, and vice versa) staining, this category reduced to 2.2%.@*Conclusions@#Among both males and females, SCC which is highly related to smoking was the most common histologic type of lung cancer. Our results showed that an approach of using only a two-antibody panel (p63 and TTF-1) might help in the reduction of diagnostic category of NSCLC-NOS significantly.

Can p63 serve as a biomarker for diagnosing giant cell tumor of bone? A systematic review and meta-analysis

ABSTRACT BACKGROUND: Tumor protein p63 (p63) has been reported to be highly expressed in giant cell tumor of bone (GCTB). Whether p63 can be treated as a diagnostic marker for GCTB remains unclear. OBJECTIVE: We conducted a meta-analysis to evaluate the applicability of p63 in diagnosing GCTB. DESIGN AND SETTING: Systematic review and meta-analysis carried out in a public hospital, Hong Kong, China. METHODS: We searched PubMed, EMBASE and the Cochrane Library from inception to April 30, 2019. Literature in English or Chinese about the differential diagnosis of GCTB using p63 were included. ­Animal experiments, reviews, correspondence, case reports, expert opinions and editorials were excluded. Studies were also excluded if they did not provide sufficient information to construct a 2 × 2 contingency table. We calculated individual and pooled sensitivities and specificities. We used I² as an indicator of heterogeneity. RESULTS: Out of 88 records identified, 8 articles on 788 GCTB patients fulfilled the inclusion criteria and were included in the present analysis. Bivariate analyses yielded a pooled mean sensitivity of 0.87 (95% confidence interval, CI, 0.72-0.95) and specificity of 0.71 (95% CI, 0.56-0.82) for using p63 as a biomarker in diagnosing GCTB. The area under the receiver operating characteristic curve was 0.86 (95% CI, 0.82-0.88). CONCLUSION: p63 is a helpful indicator in diagnosing GCTB due to its high sensitivity and specificity. ­Nonetheless, the results need to be carefully interpreted based on other diagnostic methods such as imaging. SYSTEMATIC REVIEW REGISTRATION: 164115 (PROSPERO registration number)