Heat stress affects mammary metabolism by influencing the plasma flow to the glands

Background Environmental heat stress(HS)can have detrimental effects on milk production by compromising the mammary function.Mammary plasma flow(MPF)plays a crucial role in nutrient supply and uptake in the mam-mary gland.In this experiment,we investigated the physiological and metabolic changes in high-yielding cows exposed to different degrees of HS:no HS with thermal-humidity index(THI)below 68(No-HS),mild HS(Mild-HS,68≤THI≤79),and moderate HS(Mod-HS,79<THI≤88)in their natural environment.Our study focused on the changes in blood oxygen supply and mammary glucose uptake and utilization.Results Compared with No-HS,the MPF of dairy cows was greater(P<0.01)under Mild-HS,but was lower(P<0.01)in cows under Mod-HS.Oxygen supply and consumption exhibited similar changes to the MPF under different HS,with no difference in ratio of oxygen consumption to supply(P=0.46).The mammary arterio-vein differences in glucose concentration were lower(P<0.05)under Mild-and Mod-HS than under no HS.Glucose supply and flow were significantly increased(P<0.01)under Mild-HS but significantly decreased(P<0.01)under Mod-HS compared to No-HS.Glucose uptake(P<0.01)and clearance rates(P<0.01)were significantly reduced under Mod-HS compared to those under No-HS and Mild-HS.Under Mild-HS,there was a significant decrease(P<0.01)in the ratio of lac-tose yield to mammary glucose supply compared to that under No-HS and Mod-HS,with no difference(P=0.53)in the ratio of lactose yield to uptaken glucose among different HS situations.Conclusions Degrees of HS exert different influences on mammary metabolism,mainly by altering MPF in dairy cows.The output from this study may help us to develop strategies to mitigate the impact of different degrees of HS on milk production.

A cell transcriptomic profile p ovides insights into adipocytes of porcine mammary gland across development

Background Studying the composition and developmental mechanisms in mammary gland is crucial for healthy growth of newborns. The mammary gland is inherently heterogeneous, and its physiological function dependents on the gene expression of multiple cell types. Most studies focused on epithelial cells, disregarding the role of neighboring adipocytes.Results Here, we constructed the largest transcriptomic dataset of porcine mammary gland cells thus far. The dataset captured 126,829 high-quality nuclei from physiological mammary glands across five developmental stages(d 90 of gestation, G90;d 0 after lactation, L0;d 20 after lactation, L20;2 d post natural involution, PI2;7 d post natural involution, PI7). Seven cell types were identified, including epithelial cells, adipocytes, endothelial cells, fibroblasts cells, immune cells, myoepithelial cells and precursor cells. Our data indicate that mammary glands at different developmental stages have distinct phenotypic and transcriptional signatures. During late gestation(G90), the differentiation and proliferation of adipocytes were inhibited. Meanwhile, partly epithelial cells were completely differentiated. Pseudo-time analysis showed that epithelial cells undergo three stages to achieve lactation, including cellular differentiation, hormone sensing, and metabolic activation. During lactation(L0 and L20), adipocytes area accounts for less than 0.5% of mammary glands. To maintain their own survival, the adipocyte exhibited a poorly differentiated state and a proliferative capacity. Epithelial cells initiate lactation upon hormonal stimulation. After fulfilling lactation mission, their undergo physiological death under high intensity lactation. Interestingly, the physiological dead cells seem to be actively cleared by immune cells via CCL21-ACKR4 pathway. This biological process may be an important mechanism for maintaining homeostasis of the mammary gland. During natural involution(PI2 and PI7), epithelial cell populations dedifferentiate into mesenchymal stem cells to maintain the lactation potential of mammary glands for the next lactation cycle.Conclusion The molecular mechanisms of dedifferentiation, proliferation and redifferentiation of adipocytes and epithelial cells were revealed from late pregnancy to natural involution. This cell transcriptomic profile constitutes an essential reference for future studies in the development and remodeling of the mammary gland at different stages.

Hesperidin ameliorates H_(2)O_(2)-induced bovine mammary epithelial cell oxidative stress via the Nrf2 signaling pathway

Background Hesperidin is a citrus flavonoid with anti-inflammatory and antioxidant potential. However, its protective effects on bovine mammary epithelial cells(b MECs) exposed to oxidative stress have not been elucidated.Results In this study, we investigated the effects of hesperidin on H_(2)O_(2)-induced oxidative stress in b MECs and the underlying molecular mechanism. We found that hesperidin attenuated H_(2)O_(2)-induced cell damage by reducing reactive oxygen species(ROS) and malondialdehyde(MDA) levels, increasing catalase(CAT) activity, and improving cell proliferation and mitochondrial membrane potential. Moreover, hesperidin activated the Keap1/Nrf2/ARE signaling pathway by inducing the nuclear translocation of Nrf2 and the expression of its downstream genes NQO1 and HO-1, which are antioxidant enzymes involved in ROS scavenging and cellular redox balance. The protective effects of hesperidin were blocked by the Nrf2 inhibitor ML385, indicating that they were Nrf2 dependent.Conclusions Our results suggest that hesperidin could protect b MECs from oxidative stress injury by activating the Nrf2 signaling pathway, suggesting that hesperidin as a natural antioxidant has positive potential as a feed additive or plant drug to promote the health benefits of bovine mammary.

Detection of Progesterone Receptor as a Method of Diagnosing Mammary Cancer in Female Dogs

Introduction: Canine mammary gland tumor is the most common type of neoplasia in non-ovariectomized bitches. Approximately 50% of tumors are malignant. Neoplasms originating from the mammary gland represent the most common neoplastic disease in canines in Veterinary Medicine. Aim: Relate the expression of the receptor to progesterone (PR) with the tumor stage of canine mammary carcinoma. Material and Methods: Analytical-cross-sectional study, samples of paraffinized tumor tissue obtained from 30 canine patients with breast cancer were used. The expression of PR was performed by immunohistochemical labeling, using murine anti-PR (anti-PR Biocare brand). A descriptive analysis was carried out with the results using the SPSS program. Results: The predominant histological subtype of breast cancer was tubular carcinoma with 12 patients, followed by papillary cystic carcinoma with 6 patients, solid carcinoma 5, carcinosarcoma 4 and comedocarcinoma 3. There was a significant trend between breast cancer subtypes, histological grade G1. Among the histopathological findings, the degree of invasion is related to the presence of tumor cells in adjacent lymph nodes, which is why it is a prognostic indicator. The expression of PR in the tumor tissue samples it was 42.8% positive versus 57.14% negative, of which 75% correspond to G1, 8.3% to G2 and 16.6% to G3. With respect to the relationship of the expression of PR vs type of tumor, it was found that 50% correspond to tubular carcinoma, 33.3% to papillary cystic carcinoma, 8.3% to solid carcinoma and 8.3% to comedocarcinoma. Conclusion: The hormone receptor was negative in more than half of the patients and histological grade is significantly associated with tumor subtypes, this study emphasizes the need to introduce receptor testing into our routine clinical practice to offer the best treatment for breast cancer.

Sulforaphane prevents LPS‑induced inflammation by regulating the Nrf2‑mediated autophagy pathway in goat mammary epithelial cells and a mouse model of mastitis

Background Mastitis not only deteriorates the composition or quality of milk,but also damages the health and pro-ductivity of dairy goats.Sulforaphane(SFN)is a phytochemical isothiocyanate compound with various pharmacologi-cal effects such as anti-oxidant and anti-inflammatory.However,the effect of SFN on mastitis has yet to be elucidated.This study aimed to explore the anti-oxidant and anti-inflammatory effects and potential molecular mechanisms of SFN in lipopolysaccharide(LPS)-induced primary goat mammary epithelial cells(GMECs)and a mouse model of mastitis.Results In vitro,SFN downregulated the mRNA expression of inflammatory factors(tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6),inhibited the protein expression of inflammatory mediators(cyclooxygenase-2(COX2),and inducible nitric oxide synthase(iNOS))while suppressing nuclear factor kappa-B(NF-κB)activation in LPS-induced GMECs.Additionally,SFN exhibited an antioxidant effect by increasing Nrf2 expression and nuclear translocation,up-regulating antioxidant enzymes expression,and decreasing LPS-induced reactive oxygen species(ROS)produc-tion in GMECs.Furthermore,SFN pretreatment promoted the autophagy pathway,which was dependent on the increased Nrf2 level,and contributed significantly to the improved LPS-induced oxidative stress and inflammatory response.In vivo,SFN effectively alleviated histopathological lesions,suppressed the expression of inflammatory factors,enhanced immunohistochemistry staining of Nrf2,and amplified of LC3 puncta LPS-induced mastitis in mice.Mechanically,the in vitro and in vivo study showed that the anti-inflammatory and anti-oxidative stress effects of SFN were mediated by the Nrf2-mediated autophagy pathway in GMECs and a mouse model of mastitis.Conclusions These results indicate that the natural compound SFN has a preventive effect on LPS-induced inflam-mation through by regulating the Nrf2-mediated autophagy pathway in primary goat mammary epithelial cells and a mouse model of mastitis,which may improve prevention strategies for mastitis in dairy goats.

Current status of outcome reporting in randomized controlled trials of traditional Chinese medicine for mammary gland hyperplasia: A systematic review

Objective:To assess outcome indicators in clinical trials and provide a reference for establishing a core outcome set to treat hyperplasia of mammary gland(HMG)with traditional Chinese medicine(TCM).Methods:Eight online databases were searched from their inception to December 31,2022,to assess outcomes reported in randomized controlled trials(RCTs)of HMG treated with TCM.The quality of the included studies was assessed according to the Cochrane Risk of Bias Assessment Tool.All outcomes were extracted,classified,and described.Results:A total of 8249 articles were initially retrieved.Of these,70 articles were eligible and involved 10618 participants with HMG.A total of 17 outcome indicators with a frequency of 271 times were involved and were collected according to six outcome domains.Conclusions:The core outcomes of RCTs of HMG treated with TCM are large and divergent.There are problems in evaluation standards,primary and secondary outcomes,TCM characteristic indicators,long-term prognosis,and standardization of reporting.It is recommended to strengthen the trial design and actively construct the core outcome sets with TCM characteristics for HMG.

The LXRB-SREBP1 network regulates lipogenic homeostasis by controlling the synthesis of polyunsaturated fatty acids in goat mammary epithelial cells

Background:In rodents,research has revealed a role of liver X receptors(LXR) in controlling lipid homeostasis and regulating the synthesis of polyunsaturated fatty acids(PUFA).Recent data suggest that LXRB is the predominant LXR subtype in ruminant mammary cells,but its role in lipid metabolism is unknown.It was hypothesized that LXRB plays a role in lipid homeostasis via altering the synthesis of PUFA in the ruminant mammary gland.We used overexpression and knockdown of LXRB in goat primary mammary epithelial cells(GMEC) to evaluate abundance of lipogenic enzymes,fatty acid profiles,content of lipid stores and activity of the stearoyl-Co A desaturase(SCD1) promoter.Results:Overexpression of LXRB markedly upregulated the protein abundance of LXRB while incubation with si RNA targeting LXRB markedly decreased abundance of LXRB protein.Overexpression of LXRB plus T0901317(T09,a ligand for LXR) dramatically upregulated SCD1 and elongation of very long chain fatty acid-like fatty acid elongases 5–7(ELOVL 5–7),which are related to PUFA synthesis.Compared with the control,cells overexpressing LXRB and stimulated with T09 had greater concentrations of C16:0,16:1,18:1n7,18:1n9 and C18:2 as well as desaturation and elongation indices of C16:0.Furthermore,LXRB-overexpressing cells incubated with T09 had greater levels of triacylglycerol and cholesterol.Knockdown of LXRB in cells incubated with T09 led to downregulation of genes encoding elongases and desaturases.Knockdown of LXRB attenuated the increase in triacylglycerol and cholesterol that was induced by T09.In cells treated with dimethylsulfoxide,knockdown of LXRB increased the concentration of C16:0 at the expense of C18:0,while a significant decrease in C18:2 was observed in cells incubated with both si LXRB and T09.The abundance of sterol regulatory element binding transcription factor 1 precursor(p SREBP1) and its mature fragment(n SREBP1) was upregulated by T09,but not LXRB overexpression.In the cells cultured with T09,knockdown of LXRB downregulated the abundance for p SREBP1 and n SREBP1.Luciferase reporter assays revealed that the activities of wild type SCD1 promoter or fragment with SREBP1 response element(SRE) mutation were decreased markedly when LXRB was knocked down.Activity of the SCD1 promoter that was induced by T09 was blocked when the SRE mutation was introduced.Conclusion:The current study provides evidence of a physiological link between the LXRB and SREBP1 in the ruminant mammary cell.An important role was revealed for the LXRB-SREBP1 network in the synthesis of PUFA via the regulation of genes encoding elongases and desaturases.Thus,targeting this network might elicit broad effects on lipid homeostasis in ruminant mammary gland.

Fractional flow reserve measured via left internal mammary artery after coronary artery bypass grafting:Two case reports

BACKGROUND The fractional flow reserve(FFR)has made the treatment of coronary heart disease more precise.However,there are few reports on the measurement of FFR via the left internal mammary artery(LIMA).Herein,we described the determination of further treatments by measuring FFR via the LIMA in 2 cases after coronary artery bypass grafting(CABG).CASE SUMMARY Case 1 was a 66-year-old male who was admitted due to“chest tightness after CABG.”The patient underwent CABG 7 years prior due to coronary heart disease.Coronary artery angiography showed complete occlusion of the left anterior descending artery(LAD),and subtotal occlusion of the third segment of the right coronary artery.On arterial angiography,there was 85%stenosis at the distal end of the anastomosis of the LIMA-LAD graft.FFR via LIMA was determined at 0.75.Thus,balloon dilation was performed in Case 1.FFR after balloon dilation was 0.94.Case 2 was a 60-year-old male who was admitted due to“chest tightness after CABG.”The patient underwent CABG 6 years prior due to coronary heart disease.There was 60%segmental stenosis in the middle segment of LAD and 75%anastomotic stenosis.FFR measured via LIMA was 0.83(negative);thus the intervention was not performed.Case 2 was given drug treatments.At the 3-mo follow-up,there was no recurrence of chest tightness or shortness of breath in both cases.They are currently under continual follow-up.CONCLUSION We provided evidence that FFR measurement via grafted blood vessels,especially LIMA,after CABG is a good method to determine the intervention course.

Primary Culture of Bovine Mammary Epithelial Cells

[ Objective] To investigate the feasibility of the primary culture of bovine mammary epithelial cells in biochemical incubator. [ Method] In vitro, bovine mammary epithelial cells were isolated and cultured by the tissue explant method in order to investigate the optimal culture conditions. The morphology observation and identification of the cultured cells were performed by inverted microscope observation, Giemsa staining and cytokeratin immunohistochemistry. [ Result] Observed with inverted microscope, most of the bovine mammary epithelial cells were polygonal and displayed typical slabstone-like appearance. As it can be seen from cell staining results, the cell body was big and the nucleus was stained dark blue and was round or oval in shape, with clearly visible nucleoli, generally 2 -4 nucleoli. The tissue-specific expression of cytokeratin 14 and cytokeratin 18 genes in mammary epithelial cells was identified by cytokeratin immunohistochemistry. [ Conclusion] Primary bovine mammary epithelial cells were successfully cultured in biochemical incubator.

Identification of Differentially Expressed MicroRNAs During the Development of Chinese Murine Mammary Gland

MicroRNAs （miRNAs） are endogenous -22 nucleofide-long noncoding RNAs. In this study, to investigate miRNA expression profiles and their functions in mammary gland development, we have used microarray as well as qRT-PCR, to analyze the miRNA expression changes along the murine mammary cycle during pregnancy, particularly on transition from pregnancy to lactation. It shows that every developmental stage of the mammary gland has its own mjRNA expression pattern. Compared with virgin and involution, some miRNAs such as miR-138 and miR-431 are downregulated, whereas, some miRNAs such as miR-133 and miR-133a-133b are upregulated during pregnancy and lactation. These results indicate that miRNAs are functionally involved in mammary gland development.

Off-pump coronary artery bypass grafting using a bilateral internal mammary artery Y graft

Objective To evaluate the outcome of off-pump coronary artery bypass grafting （OPCABG） using a bilateral internal mammary artery （BIMA） Y configuration graft to achieve total arterial myocardial revascularization. Methods From October 2002 to December 2008, 208 patients （196 males and 12 females） underwent OPCABG using a BIMA Y configuration graft. The average age of the patients was 56.5 ± 11.3 years, with an age range of 33-78 years. A total of 167 （80.2%） cases had triple-vessel disease. Left main stem disease was found in 33 （15.9%） cases, and double-vessel disease was found in 8 （3.9%） cases. The semi-skeletonization technique was used to harvest the two internal mammary arteries （IMAs）, and then the free right internal mammary artery was connected end-to-side to the left internal mammary artery （LIMA） in situ to complete the Y configuration graft. Off-pump and sequential anastomosis methods were used to perform coronary artery bypass surgery in all patients. Graft patency was assessed intm-operatively with the HT311 transit time flowmeter. Results A total of 728 distal anastomoses were performed in 208 patients, with the average being 3.5± 1.3 per person. No one died or experienced recurrent angina within 30 days after the operation. Conclusions OPCABG using the BIMA Y graft was safe and effective to achieve total arterial revascularization. This method avoids surgical operation on the ascending aorta and other incisions.

Optimization of Parameters of Exogenous Gene Mediated by Liposome to Transfect Yak Mammary Epithelial Cells in Vitro

[Objective] The aim of this study was to optimize conditions of exogenous gene mediated by liposome to transfect yak mammary epithelial cells in Vitro.[Method] Yak mammary epithelial cells were isolated and cultivated in Vitro by the methods of collagenase digestion and tissue adhesion.The expression of cytokeratin in yak mammary epithelial cell was detected by immunocytochemistry technique.With green fluorescence protein as the report gene,yak mammary epithelial cells were transfected with exogenous gene m...

Expression of Human GCSF in Mammary Gland of Mice by Injection of Plasmid DNA

The vectors carrying the genes coding for the proteins of interest are of unpredictable efficiency in transgenic animals. The expression vector of mammary gland (pINGG) containing GCSF genomic DNA was injected into mouse mammary gland, and expression was detected in the milk of mice. The result showed that mammary gland injection method could provide a convenient transient system to confirm vector validity.

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.

Extramammary Paget’s Disease Covered the Left Nipple and Areola

We present a case of a 71-year-old woman suffering from mammary Paget,s disease and having a 10-years history of an irregular, widespread erosion accompanied by itching and burning on the skin of her left chest, extending to the breast. The erosion had steadily enlarged and had become increasingly tender. The nipple and areola of the left breast disappeared and could not be recognized. No abnormality of the right nipple, areola, and covering skin and no supernumerary nipple were seen. Mammography and ultrasonography could not be performed because of severe pain and erosive wetness. Histopathology of a surgical biopsy specimen showed epidermal infiltration by large, round, clear atypical cells scattered individually or in small clusters and distributed horizontally throughout the epidermis. The cytoplasm of these large cells was pale and vacuolated and was equivalent to that in nipple cells in Paget,s disease, and a diagnosis of mammary Paget,s disease was made. We performed total mastectomy of the left breast with wide excision of the Paget lesion of the left chest and axillary lymph node sampling. Histological examination of the specimen showed typical distribution of Paget,s cells;however no ductal carcinoma in situ was found in the mammary ducts and invasive growth was not recognized beyond the basal membrane of the lesion. From this evidences, we established a diagnosis of large, irregulaly shaped unusual mammary Paget,s disease, not of breast cancer origin, covering the left breast, areola, and nipple.

Amino acids and mammary gland development:nutritional implications for milk production and neonatal growth

Milk is synthesized by mammary epithelial cells of lactating mammals. The synthetic capacity of the mammary gland depends largely on the number and efficiency of functional mammary epithelial cells. Structural development of the mammary gland occurs during fetal growth, prepubertal and post-pubertal periods, pregnancy, and lactation under the control of various hormones （particularly estrogen, growth hormone, insulin-like growth factor-I, progesterone, placental lactogen, and prolactin） in a species- and stage-dependent manner. Milk is essential for the growth, development, and health of neonates. Amino acids （AA）, present in both free and peptide-bound forms, are the most abundant organic nutrients in the milk of farm animals. Uptake of AA from the arterial blood of the lactating dam is the ultimate source of proteins （primarily 13-casein and a-lactalbumin） and bioactive nitrogenous metabolites in milk. Results of recent studies indicate extensive catabolism of branched-chain AA （leucine, isoleucine and valine） and arginine to synthesize glutamate, glutamine, alanine, aspartate, asparagine, proline, and polyamines. The formation of polypeptides from AA is regulated not only by hormones （e.g., prolactin, insulin and glucocorticoids） and the rate of blood flow across the lactating mammary gland, but also by concentrations of AA, lipids, glucose, vitamins and minerals in the maternal plasma, as well as the activation of the mechanistic （mammalian） target rapamycin signaling by certain AA （e.g., arginine, branched-chain AA, and glutamine）. Knowledge of AA utilization （including metabolism） by mammary epithelial cells will enhance our fundamental understanding of lactation biology and has important implications for improving the efficiency of livestock production worldwide.

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.

Milk production and composition and metabolic alterations in the mammary gland of heat-stressed lactating dairy cows

This experiment was conducted to investigate the effects of heat stress(HS) on the feed intake, milk production and composition and metabolic alterations in the mammary gland of dairy cows. Twenty Holstein cows were randomly assigned to one of two treatments according to a completely randomized design. Half of the cows were allocated to the HS group in August(summer season), and the other half were assigned to the HS-free group in November(autumn season). HS reduced(P<0.01) dry matter intake(DMI), milk yield, milk protein and milk urea nitrogen(MUN) of cows compared with HSfree control, but increased(P<0.01) milk somatic cell counts(SCC). We determined the HS-induced metabolic alterations and the relevant mechanisms in dairy cows using liquid chromatography mass spectrometry combined with multivariate analyses. Thirty-four metabolites were identified as potential biomarkers for the diagnosis of HS in dairy cows. Ten of these metabolites, glucose, lactate, pyruvate, lactose, β-hydroxybutyrate, citric acid, α-ketoglutarate, urea, creatine, and orotic acid, had high sensitivity and specificity for HS diagnoses, and seven metabolites were also identified as potential biomarkers of HS in plasma, milk, and liver. These substances are involved in glycolysis, lactose, ketone, tricarboxylic acid(TCA), amino acid and nucleotide metabolism, indicating that HS mainly affects lactose, energy and nucleotide metabolism in the mammary gland of lactating dairy cows. This study suggested that HS might affect milk production and composition by affecting the feed intake and substance metabolisms in the mammary gland tissue of lactating dairy cows.

Phytochemical analysis and anticancer capacity of Shemamruthaa,a herbal formulation against DMBA-induced mammary carcinoma in rats

Objective:To investigate the bioactive-constituents of Shemamruthaa(SM),a herbal combination and its therapeutic effects on the mitochondrial functions with reference to lipid peroxidation(LPO),antioxidant status,citric acid cycle enzymes and electron transport chain enzymes in mammary tissues of 7,12-dimethylbenz(a)-anthracene(DMBA)induced mammary carcinoma in rat model.Methods:Adult Female Sprague-Dawley rats were used for the study and were divided into four groups.CroupⅠserved as control and CroupⅡrats were induced mammary carcinoma by administration of DMBA(25 mg/kg b.w.)orally.The normal and cancer-induced rats(GroupⅢ)were treated with SM(400 mg/kg b.w./day)orally by gastric incubation for 14days.CroupⅣrats served as SM-treated control animals.Results:Cancer-induced rats showed a considerably increased level of LPO with concomitant decreased levels of antioxidants,citric acid cycle enzymes,electron transport chain enzymes and cytochrome contents in the mammary tissue.Treatment with SM brought back the aforementioned biochemical parameters to near normal.Conclusions:From the results,it can be inferred that Shemamruthaa possesses significant anticancer effect through its role in attenuation of LPO,prevention of membrane damage and restoring membrane integrity.

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.