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.

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.

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.

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.

Effects of dietary valine supplementation during late gestation on the reproductive performance and mammary gland development of gilts

Background:Mammary gland development during late gestation in gilts is a major factor that alters the composition of colostrum and growth performance of piglets.Plasma valine is taken up and metabolized extensively by the mammary gland;however,the effects of valine on mammary gland development during late gestation are still unclear.Thirty primiparous gilts were divided into three treatment groups(n=10)and received one of the three diets starting on day 75 of gestation until the day of farrowing.The total dietary valine to lysine ratio of the three diets was 0.63(LV),0.73(MV),and 0.93(HV),respectively.Results:Dietary valine supplementation during late gestation did not affect(P>0.05)the litter size and weight at farrowing;however,the piglet weight and average daily gain at weaning were linearly increased(P<0.05)as the dietary valine increased.The highest piglet weight at weaning was observed when the gilts were provided the HV diet.Dietary valine supplementation linearly elevated(P<0.05)protein,fat and solids-not-fat and some free amino acids content in colostrum.The concentration of prolactin in plasma of gilts was linearly increased in response to valine supplementation at days 1 and 10 of lactation(P<0.05).Furthermore,with increasing dietary valine allowance,a linear increase(P<0.05)was observed in the area of the lumen of alveolus and the content of DNA,RNA,and total protein in the mammary tissues at day 1 of lactation.Moreover,the protein expression of cyclin D1,p-mTOR,p-S6,and p-4EBP1 was also linearly increased(P<0.05)in the mammary tissue at day 1 of lactation.However,no difference(P>0.05)was observed in the indices related to mammary development and the mTOR signaling pathway at day 21 of lactation.Conclusion:The results revealed that increasing the total dietary valine to lysine ratio to 0.93 during late gestation significantly enhances the piglet weight and average daily gain at weaning probably due to improved development of mammary gland.

The Effect of Administration of Rutin on Plasma Levels of Estrogen, Prolactin, Growth Hormone and Gene Expression of Their Receptors in Mammary Glands in Ovariectomized Rats

The development of mammary glands, endocrine hormone concentrations and the gene expression of related receptors were measured in ovariectomized virgin rats after adminstration of an estrogen-like plant extract, rutin. Thirty-two ovariectomized virgin Wistar rats were randomly assigned to 4 treatments with 8 animals each： gastric infusion of 2 mL normal saline per unovariectomized rat per day （Sham）, gastric infusion of 2 mL normal saline per ovariectomized rat per day （Ova）, gastric infusion of 60 mg rutin kg-1 body weight （BW） per ovariectomized rat per day （Ova＋Rut）, or intramuscular injection of 60 ug estradiol kg-1 BW per ovariectomized rat weekly （Ova＋Est）. Samples of blood and mammary glands were harvested to determine the levels of estrogen （E2）, prolactin （PRL） and growth hormone （GH）, and the gene expression of estrogen receptors （ER）, prolactin receptors （PRLR） and growth hormone receptors （GHR） with radioimmunoassy （RIA） and RT-PCR technology, respectively. The E2 concentration in plasma and gland tissues from the rats of Ovx＋Rut or Ovx＋Est was higher than that of Ovx （P〈0.05）, but the plasma E2 concentration from the rats of Ovx＋Rut was lower than that of Sham （P〈0.05）. The order of the PRL concentration in plasma and gland tissues was Ovx〈Ovx＋Rut〈Ovx＋Est 〈Sham, and the difference in each treatment （P〈0.05）. The plasma GH concentration was lower in Ovx than in Ovx＋Rut or Ovx＋Est, and lower in Ovx＋Rut than in Sham （P〈0.05）. The GH concentration in gland tissues was lower in Ovx than in Ovx＋Rut or Ovx＋Est （P〈0.05）, and lower in Ovx＋Rut than in Sham （P〈0.05）. The gene expression of ER in gland tissues was increased in an order as Ovx〈Ovx＋Rut〈Ovx＋Est〈Sham （P〈0.05）, and PRLR, GHR showed the same trend. In conclusion, adminstration of rutin increased the E2 concentration in plasma and mammary glands, promoted pituitary PRL and GH release, up-regulated the gene expression of ER, PRLR and GHR, and stimulated mammary development in ovariectomized virgin rats.

Mammary gland growth and vascularity at parturition and during lactation in primiparous ewes fed differing levels of selenium and nutritional plane during gestation

Background： Objectives were to examine the effects of selenium （Se） supply and maternal nutritional plane during gestation on mammary gland growth, cellular proliferation, and vascularity at parturition and d 20 of lactation. Rambouillet primiparous ewes （n = 84） were allocated to treatments in a 2 x 3 factorial. Factors were dietary Se （adequate Se [ASe, 11.5 μg/kg BW] or high Se [HSe, 77.0 μg/kg BVV]） and nutritional plane （60% IRES], 100% [CON], or 140% [EXC]）. At parturition, lambs were removed and 42 ewes （7/treatment） were necropsied. Remaining ewes were fed a common diet meeting requirements for lactation and mechanically milked twice daily until necropsy on d 20. At both necropsy periods, mammary glands were dissected and tissues harvested. Samples were analyzed for RNA, DNA, and protein content, cell proliferation, and vascularity. Where interactions were present （P 〈 0.05）, least squares means from the highest-order interaction are presented. Results： Final body weight of ewes was least （P 〈 0.002） in RES, intermediate for CON, and greatest for EXC, regardless of stage of the ewe at necropsy （parturition or d 20 of lactation）. In ewes necropsied at parturition, mammary glands were heavier （P = 0.02） in EXC compared to RES, with CON intermediate. Concentration of RNA （rag/g） was decreased （P= 0.01） in EXC compared to CON at parturition. There was a tendency （P= 0.07） for a Se by nutrition interaction in percentage of cells proliferating where ASe-EXC ewes had greater （P_〈 0.02） number of proliferating cells then all other treatments. Mammary vascular area tended （P = 0.08） to be affected by a Se by nutrition interaction where ASe-CON had less （P= 0.007） vascular area than HSe-CON ewes. In ewes necropsied at d 20 of lactation, the number of alveoli per area was decreased （P 〈- 0.05） in RES compared to CON and EXC-fed ewes. Conclusions： Results of this study indicate that proper maternal nutritional plane during gestation is important for mammary gland development, even out to d 20 of lactation.

AMPK-mTOR pathway is involved in glucose-modulated amino acid sensing and utilization in the mammary glands of lactating goats

Background:The local supply of energy-yielding nutrients such as glucose seems to affect the synthesis of milk components in the mammary gland(MG).Thus,our study was conducted to investigate the effects of locally available MG glucose supply(LMGS)on amino acid(AA)sensing and utilization in the MG of lactating dairy goats.Six dosages of glucose(0,20,40,60,80,and 100 g/d)were infused into the MG through the external pudendal artery to investigate the dose-dependent changes in mammary AA uptake and utilization(Exp.1)and the changes in mRNA and protein expression of the AMPK-mTOR pathway(Expt.2).Results:In Exp.1,total milk AA concentration was highest when goats were infused with 60 g/d glucose,but lower when goats were infused with 0 and 100 g/d glucose.Increasing LMGS quadratically changed the percentages ofαS2-casein andα-lactalbumin in milk protein,which increased with infusions from 0 to 60 g/d glucose and then decreased with infusions between 60 and 100 g/d glucose.The LMGS changed the AA availability and intramammary gland AA utilization,as reflected by the mammary AA flux indexes.In Exp.2,the mRNA expression of LALBA in the MG increased quadratically with increasing LMGS,with the highest expression at dose of 60 g/d glucose.A high glucose dosage(100 g/d)activated the general control nonderepressible 2 kinase,an intracellular sensor of AA status,resulting in a reduced total milk AA concentration.Conclusions:Our new findings suggest that the lactating MG in dairy goats may be affected by LMGS through regulation of the AA sensory pathway,AA utilization and protein synthesis,all being driven by the AMPK-mTOR pathway.

Role of miRNA in Mammary Gland Development and Lactation

miRNA can regulate development and milk yield of the mammary gland through epigenetic mechanism, miRNA can directly and indirectly modulate the activity of the epigenetic machinery, target genes through post-inhibition of translation initiation, mediate miRNA decay, target genes and inhibit the positive regulation, regulate tone modification, and regulate DNA methylation of target genes. Here we reviewed the role of miRNAs in mammary gland development and lactation. Researching miRNA in mammary gland development and lactation process, and understanding the response of the epigenetic mechanisms to external stimuli will be an important necessity to devise new technologies for maximizing their activity and milk production in the dairy cow.

Effect of Lirukang Oral Liquid(利乳康口服液) on Neurotransmitters and Estrogen in Rats with Hyperplasia of Mammary Gland

Objective: To investigate the effects of Lirukang oral liquid (LRK, 利乳康口服液) on release of neurotransmitter in rats with hyperplasia of mammary glands (HMG) and to explore its mechanism. Methods: Sixty rats were divided into six groups, the normal control group, the model control group, the large dosage (3.6g/kg) and the small dosage (1.8g/kg ) LRK groups, the Ruzengning (乳增宁, RZN, 2.5g/kg) group and the tamoxifen (TAM, 5mg/kg) group, 10 in each group. Except those in the normal control group, all the animals were made into rat model of HMG by intraperitoneal injection of estradiol benzoate. Levels of dopamine (DA) and 5-hydroxytryptamine (5-HT) in hypothalamus and mammary gland in rats were detected by fluorescence luminosity assay, and level of prolactin (PRL) in serum was detected by radioimmunoassay. Results: In the model group, the level of DA reduced significantly ( P<0. 01), and 5-HT and PRL increased obviously ( P<0.01). Compared with the model group, the LRK groups of both dosages and the TAM group had their level of DA significantly increased (P<0. 01), and level of 5-HT significantly decreased ( P<0.01). The serum PRL in both LRK groups was significantly decreased ( P<0. 01). No obvious changes in DA, 5-HT and PRL were found in the RZN group. Conclusion: LRK and TAM have similar effects in regulating the release of neurotransmitter in hypothalamus and mammary gland and serum content of estrogen in the animal models of HMG.

The Expression of the IGF Family During Mouse Mammary Gland Development

This study was to determine the patterns and levels of IGF family members＇ expression during postnatal mammary gland development. The authors investigated the protein expression profile of the major components of the IGF axis in murine mammary glands. All the proteins examined, IGF- Ⅰ, IGF- Ⅱ, and IGF- Ⅰ receptor （IGF-Ⅰ R） were expressed at greatly different levels and displayed unique expression profiles. IGF- Ⅱ and IGF- ⅠR were always expressed at significantly higher levels than IGF- Ⅰ. IGF- Ⅰwas localized in adipocytes as well as the epithelial and stromal compartments, but just distinctly expressed where mammary cells aggregated to form ducts, in virgins. The IGF- Ⅱ was localized only on the basal layer epithelial cell membranes of ducts and alveoli, with a peak level on the initiation of lactation. The higher level of IGF- ⅠR compared with IGF- Ⅰ was also found in adipocytes as well as in the epithelial and stromal compartments, especially during pregnancy and late lactation. The IGF- Ⅰ R pathway was obviously significant for the development of the mammary parenchyma and stroma. Overall, the comparison of the expression profiles of these different proteins would strongly suggest that they were likely to have different functions throughout the mammary gland development, and it also highlighted the potential interactions and coregulation of the members of this axis. It seems that IGF- Ⅱ was the major local modulator rather than IGF- Ⅰ by an IGF- Ⅰ R-independent pathway, especially for initiation of lactation. This study has demonstrated the importance and complexity of the IGF axis during mammary gland development and provides a valuable resource for future research in this area.

Differential Acupuncture Treatment of Hyperplasia of Mammary Glands

Hyperplasia of mammary glands is a common disease in the middle-aged women, and it is a precancerous lesion of mammary glands. For many years, the author has used Rugen (ST 18) of the Stomach Meridian of Foot-Yangming as the main point withcertain auxiliary points chosen on basis of the differentiation types to treat the disease and obtained satisfactory therapeutic effects. A report follows. ……

Core acupoints of acupuncture for hyperplasia of mammary gland: a network analysis

Backgroud:Clinical studies on acupuncture treatment of hyperplasia of mammary gland(HMG)have proved its effectiveness,but most studies have paid little attention to acupoints prescription and acupoint compatibility.The clinical prescription is not identical,the curative effect also has the difference.Therefore,through data mining and network analysis,this study explored the core acupoints and the compatibility law of acupoints in acupuncture treatment of HMG.Methods:To search and select qualified literature according to inclusion and exclusion criteria for relevant clinical research literature on acupuncture treatment of HMG in CNKI,VIP database,WanFang database and PubMed,etc.Then extract relevant information and establish a database.Using the method of statistical and complex network analysis,this paper studies the core acupoints and the law of acupoint compatibility.Results:A total of 104 Chinese literatures and 0 English literatures were included and 106 acupuncture prescriptions were extracted.The core acupoints in the treatment of HMG are Danzhong(CV 17),Wuyi(ST 15),Zusanli(ST 36),Jianjing(GB 21).Danzhong(CV 17)and Zusanli(ST 36),Danzhong(CV 17)and Wuyi(ST 15),Jianjing(GB 21)and Tianzong(SI 11),Jianjing(GB 21)and Wuyi(ST 15)have the highest correlation degree.The method of acupoint matching mainly consists of local-remote acupoints,upper-lower acupoints and front-rear acupoints.Conclusion:The results of a network analysis substantially accord with the general rules of acupuncture theories in traditional Chinese medicine,able to reflect the points-selection principles and features in acupuncture treatment of HMG and provide evidence for the acupoints selection in the treatment of HMG in acupuncture clinic.

Regulation of leptin in involution of mammary gland

Leptin, a protein hormone produced and secreted predominantly by white adipose tissue, has a critical role in the regulation and coordination of energy metabolism. Leptin is produced in the mammary gland by the fat tissue or by the mammary epithelium. In vitro study has shown that leptin triggers apoptosis in mammary epithelial cells. Mammary gland involution is characterized by extensive apoptosis of the epithelial cells. At the onset of involution, STAT3 is specifically activated.Various studies show that leptin act as a paracrine and autocrin factor to influence mammary epithelial cell proliferation and differentiation. This paper reviewed the function of leptin to the involution of mammary gland.

Potential Genes for Regulation of Milk Protein Synthesis in Dairy Goat Mammary Gland

The lactating mammary gland is a prodigious protein-producing factory, but the milk protein synthesis mechanisms are not well understood. The major objective of this paper was to elucidate which genes and pathways were involved in the regulation of milk protein synthesis in the dairy goat mammary gland. Total 36 primiparous Guanzhong dairy goats were allotted in 12 groups according to their mammary development stages： days 90 and 150 of virgin, days 30, 90, and 150 of pregnancy, days 1, 10, 35, and 60 of lactation and days 3, 7, and 21 of involution （three animals per group）. Mammary tissue RNA was isolated for quantitative real- time RT-PCR of four casein genes alpha-s 1 casein （CSN 1S 1 ）, alpha-s2 casein （CSN 1 S2）, beta-casein （CSN2） and casein kappa （CSN3）, four whey protein genes lactoglobulin （LGB）, laetalbumin （LALBA）, laetofarrin （LTF）, and Whey acidic protein （WAP） and the genes which were potentially to regulate dairy goat milk protein synthesis at the level of transcription or translation [prolactin receptor （PRLR）, AKT1, signal transducers and activators of transcription 5 （STAT5）, E74-Like Factor 5 （ELF5）, eukaryotic translation initiation factor 4E binding protein 1 （EIF4E-BP1）, S6kinase （S6K） and caveolin 1]. The results showed that all genes were up-regulated in lactation period. The expressions of PRLR, AKT1, STAT5, ELF5, and S6K were similar to mRNA expressions of milk proteins. Our results indicated that milk protein synthesis in dairy goat mammary gland was possibly regulated by these genes.

Commercial Artificial Sweeteners Might Affect Pregnant and Nursing Mice Mammary Gland. A Histological Study

The use of artificial sweeteners as non-nutritive food additive is becoming very common nowadays. The aim of this research was to study the effect of commercial artificial sweeteners consumption on the adult mouse mammary gland in pregnancy (18-day pregnancy) and 3 weeks nursing. A commercial artificial sweetener solution was given to pregnant mice of treated groups 50 mg/kg body weight, from day one of pregnancy till the end of 3 weeks nursing. Controls were given distilled water. The histological studies of the mammary gland of treated animals showed some changes. In treated 18-day pregnant mothers;the amount of adipose cells seemed to increase and the amount of lipid droplets within the alveoli seemed to increase compared to the controls. Also alveoli in the treated mammary gland sections appeared to be larger and less in number compared to the controls. In 3 weeks treated nursing mothers the amount of milk and lipid droplets in the alveoli decreased compared to the controls. The mammary gland circumference of treated 18-day pregnant mothers was significantly smaller compared to the controls, while it was significantly larger in 3 weeks treated nursing mothers compared to the controls. These results show that artificial sweeteners disturbed the arrangement of the histological structure in the mammary gland of pregnant and nursing mice. Awareness should be raised to restrict the use of artificial sweeteners to people with diabetes or who are in medical need.

Cervical Thoracic Necrotizing Fasciitis with the Mammary Gland Spread of Odontogenic Origin

Necrotizing fasciitis is an uncommon infection, but potentially lethal, especially when associated with systemic disorders such as diabetes. We report the case of a 35-year-old female with uncontrolled diabetes mellitus, presenting with edema of the neck, facial and left mammary gland, secondary to untreated dental infection, progressing to a full-blown necrotizing fasciitis in a short period of time with sepsis. The patient was managed with aggressive multidisciplinary medical and surgical treatment. Despite the technologic advances in diagnosis and treatment, complications still result with astounding high mortality. Clearly, the morbidity associated to this infection, even in diabetic patients, can be minimized if an early diagnosis and effective debridement are done.

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.

Dietary supplementation of sodium butyrate enhances lactation performance by promoting nutrient digestion and mammary gland development in dairy cows

This experiment was to evaluate the influence of sodium butyrate(SB)addition on milk production,ruminal fermentation,nutrient digestion,and the development and metabolism regulation of the mammary gland in dairy cows.Forty Holstein dairy cows averaging 710±18.5 kg body weight,72.8±3.66 d in milk(DIM),and 41.4±1.42 kg/d milk production were divided into four treatments blocked by DIM and milk production.Treatments were control group,low SB,medium SB,and high SB with 0,100,200 and 300 g/d of SB addition per cow,respectively.The study lasted for 105 d.Production of milk,milk protein and lactose quadratically increased(P<0.05),while fat-corrected milk,energycorrected milk and milk fat yields linearly increased(P<0.05)with increasing SB addition.The digestibility of dietary dry matter,organic matter,and crude protein linearly increased(P<0.05),whereas the digestibility of ether extract,neutral detergent fibre,and acid detergent fibre quadratically increased(P<0.05).Ruminal pH quadratically decreased(P=0.04),while total volatile fatty acids(VFA)quadratically increased(P=0.03)with increasing SB addition.The acetic acid to propionic acid ratio increased(P=0.03)linearly due to the unaltered acetic acid molar percentage and a linear decrease in propionic acid molar percentage.Ruminal enzymatic activity of carboxymethyl-cellulase and a-amylase,populations of total bacteria,total anaerobic fungi,total protozoa,Ruminococcus albus,R.flavefaciens,Butyrivibrio fibrisolvens,Fibrobacter succinogenes,and Ruminobacter amylophilus linearly increased(P<0.05).Blood glucose,urea nitrogen,and non-esterified fatty acids linearly decreased(P<0.05),while total protein concentration linearly increased(P=0.04).Moreover,the addition of SB at 200 g/d promoted(P<0.05)mRNA and protein expression of PPARγ,SREBF1,ACACA,FASN,SCD,CCNA2,CCND1,PCNA,Bcl-2,GPR41,and the ratios of p-Akt/Akt and p-mTOR/mTOR,but decreased(P<0.05)mRNA and protein expressions of Bax,caspase-3,and caspase-9.The results suggest that milk production and milk fat synthesis increased with SB addition by stimulating rumen fermentation,nutrient digestion,gene and protein expressions concerned with milk fat synthesis and mammary gland development.