Effect of Flos Buddlejae Granules on Apoptosis Factors Bax,Bcl-2,Fas and FasL in Lacrimal Gland Cells of Castrated Male Rabbits

Objective To observe the effect of different concentration Flos Buddlejae(Mi Meng Hua,密蒙花)granules on apoptosis factors of lacrimal gland cells of castrated male rabbits,and study the treatment effects of Flos Buddlejae(Mi Meng Hua,密蒙花)granules in the dry eye model of castrated male rabbits.Methods Flos Buddlejae(Mi Meng Hua,密蒙花)raw material was made into granules.Thirty healthy adult New Zealand rabbits were randomly divided into five groups,six rabbits in each group.Group A:blank group,Group B:model group,Group C:Flos Buddlejae(Mi Meng Hua,密蒙花)granule group,Group D:placebo group,Group E:testosterone group.Except for group A,all rabbits underwent removal of bilateral testis and epididymis.Rabbits in group C were administered Flos Buddlejae(Mi Meng Hua,密蒙花)granules(100 mg/kg),three times per day;rabbits in group D were administered normal saline,three times per day.Rabbits in group E were injected with testosterone propionate(0.5 mL/kg)in the thigh muscle,every 3 days.All rabbits were tested by Schirmer I test(SIT)and tear film break-up time(BUT)before operation and 4 weeks after operation.After 4 weeks,all rabbits were sacrificed by air embolism and clipping of the lacrimal gland.Apoptosis factors,including Bax,Bcl-2,Fas and FasL of lacrimal gland cells were characterized by immunohistochemistry,and resulting data were statistically analyzed.Results(1)Comparison of SIT and BUT before and after operation:There were statistically significant differences between groups B and D(P<0.01),but not among other groups(P>0.05).(2)Comparison of apoptosis factors Bax,Bcl-2 Fas and FasL:In a comparison of groups B and D,there was no statistically significant difference after operation(P>0.05).In a comparison of the other groups,there were statistically significant differences(P<0.01).In comparisons among A,C and E groups,there were no statistically significant differences(P>0.05).Conclusion Compared with androgen,Flos Buddlejae(Mi Meng Hua,密蒙花)granules caused similar but slightly weaker depression of Bax,Fas and FasL,and increased expression of Bcl-2.

Isoflavone Ameliorates H₂O₂Induced Injury by Activating the Antioxidant System of Sow Mammary Gland Cell

The antioxidant and protective properties of a synthetic soybean isoflavone (ISO) were investigated using sow mammary gland cell. Cells were cultured with 10, 20 and 30 uM ISO, respectively, under 80 uM FeSO4·7H2O/H2O2 conditions. After 48 h of incubation, the cells in the presence of ISO were lost less compared with that of control under oxidative damage by H2O2/FeSO4;ISO decreased the cell number at G1 and G2 stages, increased the cell number at S stage (all P < 0.05), it also reduced apoptosis of the cells (P P P < 0.05). The addition of ISO significantly promoted cell proliferation (P rd to 6th days. Upon these, the activities of total superoxide dismutase (SOD), glutathione peroxidase (GPX), total antioxidant ability (T-AOC) also were increased and the activities of NADPH oxidase (NOX) decreased by ISO treatment (all P < 0.05). ISO decreased the concentration of ROS (P P < 0.05). 10 uM, 20 uM, 30 uM ISO increased the relative mRNA abundance of SOD1 (all P P P P < 0.05) and SOD3 (all P < 0.05);ISO significantly increased the relative mRNA abundance of GPX4 (all P P < 0.05), 20 uM ISO also increased the relative mRNA abundance of NOX2 (P < 0.05). It was concluded that supplementation of ISO enhanced the anti-oxidative function and prevented lipid peroxidation, possibly through the activation of the antioxidant enzymes and inhibition of cell apoptosis.

Construction of Antibacterial Peptide CecropinB Eukaryotic Recombinant Vector and Its Expression in Dairy Goat Mammary Gland Epithelial Cells

To investigate the expression of antibacterial peptide CecropinB cDNA in dairy goat mammary gland epithelial cells,the CecropinB gene was cloned and was inserted into a eukaryotic vector pECFP-C1 to construct the recombinant plasmid pECFP-B by genetic engineering technique.Recombinant plasmid pECFP-B was transfected into dairy goat mammary gland epithelial to detect the bactericidal activity of CecropinB.The expression of CecropinB was also detected.The result of RT-PCR demonstrated CecropinB gene was expressed in transfected cells.CecropinB recombinant plasmid DNA was injected into udders and CecropinB was expressed in mammary gland,exhibiting bactericidal activity to Staphylococcus aureus in vivo experiments.

Induced in vitro Expression of Human Lactoferrin in Goat Mammary Gland Epithelial Cell

[Objective]The aim of this study was to explore the technical system of induced expression in vitro of goat mammary gland epithelial cell,and evaluate expression efficiency of mammary gland specific vector and foreign protein at the cell level.[Method]Goat mammary gland epithelial cell transfected by human lactoferrin gene was inducted by culturing in DMEM/F12 medium supplemented with 5 mg/L insulin,5 mg/L prolactin and 1 mg/L hydrocortisone.Supernatant was collected per 6 hours and concentrated.Expression situation of foreign protein were detected by SDS-PAGE and Western blotting.[Result]There was target protein expression in the induced culture medium,which molecular weight was about 42 kD.[Conclusion]The method used in this study can induce goat mammary gland epithelial cell to express foreign gene,it lays a foundation for researching heterologous expression of foreign gene and producing mammary gland bioreactor.

The effects of thyroid-stimulating hormone,estradiol and prolactin on sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels

Objective The present study investigated the sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels and the effects of thyroid-stimulating hormone(TSH),estradiol(E2)and prolactin(PRL)on NIS mRNA expression in mouse lactating mammary gland cells.

Influence on Cellular Signal Transduction Pathway in Dairy Cow Mammary Gland Epithelial Cells by Galactopoietic Compound Isolated from Vaccariae segetalis

The galactopoietic mechanism of Vaccaria segetalis is still unknown. Understanding dibutyl phthalate （DBP） separated from Vaccaria segetalis on the expression of lactation signal transduction genes of mammary gland epithelial cells, including prlr, erα, akt1, socs2, pparγ and elf5, will be helpful to reveal the molecular mechanism. Western blot and qRT- PCR were used to study the change of prlr, erα, akt, socs2, pparγ, and elf5 expression at mRNA and protein level. Co- localization expression of prolactin receptor （PRLR） and estrogen receptor α （ERα） was observed by immunofluorescence; the expression changes of miRNAs （21, 125b, 143, and 195） and the secretion of β-casein and lactose were detected by qRT-PCR and RP-HPLC. The results showed that Vaccaria segetalis active compound had similar fuctions as estrogen and/or prolactin （PRL） in dairy cow mammary gland epithelial cells （DCMECs）, increased the expressions of prlr, erα, akt1, and elf5 genes, while repressed pparγ expressions. DBP promoted socs2 mRNA expression, but its protein expressions were repressed. Furthermore, both DBP and PRL could repress the expressions of miRNA-125b, miRNA-143 and miRNA- 195 in DCMECs. DBP could repress the expression of miRNA-21, while the influence of PRL on miRNA-21 was not certain. DBP could promote the lactation ability of DCMECs by regulating the ER and PRLR cellular signal transduction pathway.

Metabolic Regulation of Mammary Gland Epithelial Cells of Dairy Cow by Galactopoietic Compound Isolated from Vaccariae segetalis

In previous experiment, we isolated a compound dibutyl phthalate （DBP） from Vaccaria segetalis which had galactopoietic function on mammary gland epithelial cells of dairy cow （DCMECs）. In this experiment, we ascertained the metabolic regulation function of DBP on DCMECs. Many genes related to lactation including Stat5, AMPK, b-casein, Glut1, SREBP-1, PEPCK, and ACC were detected by real-time PCR. Furthermore, Stat5 and AMPK were detected by Western blot and immunofluorescence co-localization, respectively. The results showed that DBP stimulates the expression of Stat5 and p-Stat5, thus activates Stat5 cell signal transduction pathway and stimulates b-casein synthesis. DBP also raises the activities of Glut1 and AMPK to stimulate glucose uptake and glycometabolism and activates the expression of AMPK downstream target genes PEPCK and ACC and expression of SREBP-1 to stimulate milk fat synthesis. In addition, the activities of HK, G-6-PDH, ICDH, ATPase, and energy charges were stimulated by DBP to increase the energy metabolism level of DCMECs. The results showed DBP stimulates energy metabolism related to galactopoietic function in DCMECs.

A NEW TYPE OF INSECT EPIDERMAL GLAND CELL

Insect epidermal gland cells were reviewed and classified into 3 classes by Noirot and Quennedey. We studied the structure and physiology of the female sex pheromone glands in Clania variegata Snellen (Lepidoptera, Psychidae) and found that their gland cells did not belong to any of the gland cell classes of Noirit et al. Further results studied by SEM and TEM have confirmed that the sex pheromone gland cell is a new type of insect epidermal gland cell.

Signet-ring cell carcinoma arising from a fundic gland polyp in the stomach

Fundic gland polyps(FGPs)are currently the most common type of gastric polyps and are usually benign.However,although rare,gastric adenocarcinoma of FGP has been recently proposed as a new variant of gastric adenocarcinoma.Here we report the first case of a49-year-old woman with focal signet ring cell carcinoma that arose from an FGP of the stomach.The tumor was completely excised by endoscopic snare polypectomy.FGPs should therefore be evaluated for malignant changes although they occur rarely,if the FGP has an erosive or irregular surface.

Gastric adenocarcinoma of the fundic gland(chief cell-predominant type): A review of endoscopic and clinicopathological features

Gastric adenocarcinoma of the fundic gland(chief cellpredominant type, GA-FG-CCP) is a rare variant of welldifferentiated adenocarcinoma, and has been proposed to be a novel disease entity. GA-FG-CCP originates from the gastric mucosa of the fundic gland region without chronic gastritis or intestinal metaplasia. The majority of GA-FG-CCPs exhibit either a submucosal tumor-like superficial elevated shape or a flat shape on macroscopic examination. Narrow-band imaging with endoscopic magnification may reveal a regular or an irregular microvascular pattern, depending on the degree of tumor exposure to the mucosal surface. Pathological analysis of GA-FG-CCPs is characterized by a high frequency of submucosal invasion, rare occurrences of lymphatic and venous invasion, and low-grade malignancy. Detection of diffuse positivity for pepsinogen-I by immunohistochemistry is specific for GA-FG-CCP. Careful endoscopic examination and detailed pathological evaluation are essential for early and accurate diagnosis of GA-FG-CCP. Nearly all GA-FG-CCPs are treated by endoscopic resection due to their small tumor size and low risk of recurrence or metastasis.

Future application of hair follicle stem cells： capable in differentiation into sweat gland cells

Background Sweat glands （SGs） can not regenerate after complete destruction in the severe skin injury, so it is important to find a ideal stem cell source in order to regenerate functional SGs. Hair follicle stem cells （HFSCs） possess the obvious properties of the adult stem cells, which are multipotent and easily accessible. In this research, we attempted to direct the HFSCs suffered from the sweat gland cells （SGCs） special differentiation by a cooperative co- culture system in vitro. Methods The designed co-culture microenvironment in the transwell was consist of two critial factors： heat shocked SGCs and dermis-like mesenchymal tissue, which appeared independently in the two control groups; after induction, the purified induced SGC-like cells were transplanted into the full-thickness scalded wounds of the nude mice, after 4 weeks, the reconstructed SG-like structures were identified by immunohistochemical and immunofluorescence analysis. Results A part of HFSCs in experimental group finally expressed SGCs phenotypes, by contrast, the control group 1 which just containing dermis-like mesenchymal tissue failed and the control group 2 consisted of heat shocked SGCs was in a poor efficiency; by immunofluorescence staining and flow cytometry analysis, the expression of HFSCs special biomarkers was down regulated, instead of the positive efficiency of SGCs special antigens increased; besides, the induced SGCs displayed a high expression of ectodysplasin A （EDA） and ectodysplasin A receptor （EDAR） genes and proteins; after cell transplantation, the youngest SG-like structures formed and be positive in SGCs special antigens, which never happened in untreated wounds （P 〈0.05）. Conclusion The HFSCs are multipotential and capable in differentiating into SGCs which promise a potential stem cells reservoir for future use; our special co-culture microenvironment is promising for HFSCs differentiating; the induced SGCs are functional and could work well in the regeneration of SGs.

Bone marrow-derived mesenchymal stem cells migrate to healthy and damaged salivary glands following stem cell infusion

Xerostomia is a severe side effect of radiation therapy in head and neck cancer patients. To date, no satisfactory treatment option has been established. Because mesenchymal stem cells（MSCs） have been identified as a potential treatment modality, we aimed to evaluate stem cell distribution following intravenous and intraglandular injections using a surgical model of salivary gland damage and to analyse the effects of MSC injections on the recruitment of immune cells. The submandibular gland ducts of rats were surgically ligated. Syngeneic adult MSCs were isolated, immortalised by simian virus 40（SV40） large T antigen and characterized by flow cytometry. MSCs were injected intravenously and intraglandularly. After 1, 3 and 7 days, the organs of interest were analysed for stem cell recruitment. Inflammation was analysed by immunohistochemical staining. We were able to demonstrate that, after intravenous injection, MSCs were recruited to normal and damaged submandibular glands on days 1, 3 and 7. Unexpectedly, stem cells were recruited to ligated and non-ligated glands in a comparable manner. After intraglandular injection of MSCs into ligated glands, the presence of MSCs, leucocytes and macrophages was enhanced, compared to intravenous injection of stem cells. Our data suggest that injected MSCs were retained within the inflamed glands, could become activated and subsequently recruited leucocytes to the sites of tissue damage.

B cell receptor signaling pathway involved in benign lymphoepithelial lesions of the lacrimal gland

AIM：To detect the expression of B cell receptor signaling pathway（BCRSP） in lacrimal gland benign lymphoepithelial lesions（LGBLEL）.METHODS：Gene microarray was used to compare whole-genome expression in lacrimal gland tissues from LGBLEL patients to tissues from orbital cavernous hemangioma（control tissues）. Expression of BCRSP was confirmed by polymerase chain reaction（PCR） and immunohistochemistry. RESULTS：The expression of 22 genes of the BCRSP increased significantly in LGBLEL patients. PCR analysis showed that CD22, CR2, and BTK were all highly expressed in LGBLEL tissues. Immunohistochemical analysis showed that CR2 protein was present in LGBLEL, but CD22 and BTK proteins were negative. CR2, CD22, and BTK were not observed in the orbital cavernous hemangiomas with either PCR or immunohistochemistry. CONCLUSION：BCRSP might be involved in the pathogenesis of LGBLEL.

Mantle Cell Lymphoma in a Lacrimal Gland in a Female and a Review of the Literature

Purpose:To report a rare case of Mantle cell lymphoma in lacrimal gland and review of the literature Case report:.We report a case of a 59-year-old female who presented with an upper eyelid mass in the right eye for 3months, without pain and irrigation. A computerized tomography(CT) scan showed a mass in the bilateral lacrimal gland region, more significant in right eye. The patient underwent a lacrimal gland mass excision surgery and diagnosis of mantle cell lymphoma by histopathology..Immunochemistry for CD20, CD79 a, CD5, and Cyclin D1 was positive. She was recommended to the Shantou cancer hospital for chemotherapy.Conclusion:.Mantle cell lymphoma is a rare type of malignant lymphoma,.over expressing CD5 and cyclin D1 antigens,which distinguishes it from other B cell lymphomas.

Comparison on In-Vitro Culture Methods of Yak Endometrial Gland Epithelial Cells

[ Objective] To develop in-vitro culture methods of yak endometrial gland epithelial cells. [Method] The gland epithelial cells were isolated from yak endometrium by explant culture method and digestion culture method, respectively. [ Result] In the first method, the cells isolated from the endometrium explant could merge into monolayer after 8-d culture, and they could be purified by gradation digestion with trypsin. In the second method, the endometrium explant were first digested by collagenase II by incubation at 37℃ for 2.5 h and then further digested in fresh 2 g/L colla- genase II for another 2.5 h. The cell suspension was leached through 74 pm filter and centrifuged at 400 r/min for 5 min. Then the cell pellet was re-suspended, followed by natural sedimentation to collect purified gland epithelial cells. The isolated cells were cytokeratin-positive as detected by immunocytochemical staining, and the positive rate could reach 95%. [Conclusion] The yak endometrial gland epithelial cells can be isolated and purified by both the explant culture method and digestion culture method.

The Effects of Insulin and Prolactin on an Epithelial Cell Line from Mammary Gland of Dairy Goat

Mammary epithelial cells with lactational function can be a valuable cellular model for research of the development and regulation of the mammary gland.This paper describes some aspects of function of an epithelial cell line from the mammary gland of the dairy goat.SDS-PAGE,triglyceride and lactose content of cultured cells were used to assess synthetic function of cells and the effects of exposure to insulin and prolactin.Results show that goat mammary epithelial cells can synthesize fat,proteins and lactose when they were cultured in DMEM-F12 medium with added EGF,IGF-1,ITS and FBS.There were no obvious changes after 48h treatment with additional insulin.Prolactin added to the basal medium significantly increased synthesis of proteins and lactose.A mammary gland epithelial cell line from goats which has lactational function has been established.This outcome provides a valuable and convenient model system.

Calcium Participates in Secretion of Porphyrin from Shell Gland Epithelial Cells of Japanese Quail (<i>Coturnix coturnix japonica</i>)

To investigate factors involved in the secretion of protoporphyrin IX (PpIX), a superficial eggshell pigment, from shell gland epithelial cells of Japanese quail, we cultured cells in Ham’s F12 medium with calcium chloride and quail plasma. The addition of hormones (prostaglandin F2α, progesterone, estradiol-17β) to the medium did not change the PpIX concentration in the culture supernatant, but changing the calcium chloride (CaCl2) concentration did: a lower concentration of CaCl2 led to a higher PpIX concentration;0 mM CaCl2 enhanced the secretion of PpIX from epithelial cells prepared at 5 or 7 mM CaCl2. The result suggests that a drop in concentration of CaCl2 mimics the end of shell calcification and stimulates rapid secretion of PpIX in vivo. Bovine serum albumin was almost as effective as quail plasma for PpIX secretion in culture, and would facilitate further study of the mechanism of PpIX secretion.

<i>In Vitro</i>Culture of Shell Gland Epithelial Cells in Japanese Quail (<i>Coturnix coturnix japonica</i>)

To investigate the secretion of protoporphyrin IX (PpIX), superficial eggshell pigment, from shell gland cells of Japanese quail, the epithelial cells of the gland were collected and isolated for cultivation in vitro. An analysis of a peak for PpIX in the cells was performed using a fluorescence microplate reader. The measurement showed that PpIX has a peak of excitation wavelength at 410 nm and emission wavelength at 606 nm in the culture medium (HamF12 + 4% HCl). Volumes of PpIX in the medium after 4 hour culture of the cells were measured with a microplate reader using filter set of excitation wavelength 400/30nm and emission wavelength 620/40nm. However the cells did not secrete significantly PpIX during 4 hour incubation in this culture system, addition of quail plasma to the medium resulted in significantly higher secretion. A cultivation system in this study is able to use for the study on the mechanism of the secretion of eggshell pigment, PpIX from Japanese quail shell gland epithelial cells.

Pleomorphic Adenoma with Exuberant Squamous Metaplasia and Keratin Cysts Mimicking Squamous Cell Carcinoma in Minor Salivary Gland

Salivary gland tumors, the second most common neoplasm of the mouth after squamous cell carcinoma, account for a significant proportion of tumors of the oral and perioral regions. An unusual case of adenoma presented as a solitary intraoral palatine mass in a 32-year-old woman is reported here. The tumor was interpreted as an unusual pleomorphic adenoma because of the presence of exuberant squamous metaplasia, clinically mimicking squamous cell carcinoma. Moreover, the presence of cystic structures filled with keratinized material was also salient feature. Pleomorphic adenomas may occasionally display focal squamous metaplastic changes;when extensive, it presents the potential for misinterpretation of the histology as indicative of well-differentiated squamous cell carcinoma.

Predictors of distant metastasis in acinic cell carcinoma of the parotid gland

BACKGROUND AiCC is a primarily indolent disease process.Our aim with this study is to determine characteristics consistent with rapidly progressive AiCC of the parotid gland.AIM To report on patients with metastatic lung disease from AiCC and potential correlative factors.METHODS Single-institution retrospective review of patients treated at the University of Michigan between 2000 and 2017.Univariate analyses were performed.RESULTS A total of 55 patients were identified.There were 6 patients(10.9%)with primary AiCC of the parotid gland who developed lung metastases.The mean age at diagnosis for patients with lung metastases was 57.8 years of age,in comparison to 40.2 years for those without metastases(P=0.064).All 6 of the patients with lung metastases demonstrated gross perineural invasion intraoperatively,in comparison to none of those in the non-lung metastases cohort.Worse diseasefree and overall survival were significantly associated with gross perineural invasion,high-grade differentiation,and T4 classification(P<0.001).CONCLUSION AiCC of the parotid gland is viewed as a low-grade neoplasm with good curative outcomes and low likelihood of metastasis.With metastasis,however,it does exhibit a tendency to spread to the lungs.These patients thereby comprise a unique and understudied patient population.In this retrospective study,factors that have been shown to be statistically significant in association with worse disease-free survival and overall survival include presence of gross facial nerve invasion,higher T-classification,and high-grade disease.