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. ……

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.

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.

300 CASES OF HYPERPLASIA OF MAMMARY GLANDS TREATED WITH RED-HOT NEEDLE PLUS FILIFORM NEEDLE

Clinical observations on 300 cases of hyperplasia of mammary glands (group A) treated by red hot needle plus filiform needle are reported in the present study. "Ashi" points were used as the main acupoints. For comparison, other 300 cases (group B) were treated with routine acupuncture method, and Rugen (ST 18) was used as the main acupoint. In addition, other auxiliary acupoints were supplemented in the light of the concrete syndromes in these two groups. The treatment was given once daily, with 10 sessions being a therapeutic course. Of the 300 cases in group A, 240 were clinically cured, amounting to 80%; 58 were effective, amounting to 19.3%; 2 were failed, with a total effective rate of 99.3%. Of the 300 cases in group B, 113 were clinically cured, 165 effective, 22 were failed, with a total effective rate of 92.7%. The therapeutic effect of group A is significantly superior to that of group B (P<0.05).

CLINICAL OBSERVATION ON THE ACUPOINT INTRADERMAL EMBEDDING OF RINGHEADED THUMBTACK-NEEDLE THERAPY FOR HYPERPLASIA OF MAMMARY GLANDS

In the present paper, the author used acupoint intradermal embedding of ringheaded thumbtack-needle therapy combined with herbal medicine therapy to treat 110 cases of hyperplasia of mammary glands, and compared its therapeutic effect with that of herbal therapy alone. The observed indexes of the treatment were pain relief degree, the size of hyperplasic glands and the time of occurrence of marked effect. Following 3 courses of treatment,the results showed the therapeutic effect of the ringheaded thumbtack-needle needle plus herbal medicine therapy was superior to that of herbal medicine therapy alone in the above-mentioned 3 indexes (P ＜ 0.01 ).

ACUPUNCTURE FOR TRERTING 120 CASES OF HYPERPLASIA OF MAMMARY GLANDS

Objective: To observe the therapeutic effect of acupuncture therapy for 120 cases of hyperplasia of mammary glands. Methods: These patients were classified into liver-qi-stagnation type (n=53), phlegm-coagulation type (n=30) and Thoroughfare-Conception vessel maladjustment type (n=37). Acupoints used were Qimen (期门 LR 14), Wuyi (屋翳 ST 15), Weishu (胃俞 BL 21), etc.. Acupuncture treatment was conducted once daily and 30 treatments constituted one therapeutic course. Results: After 3 courses of treatment, of the 53, 30 and 37 cases of the liver-qi-stagnation, phlegm-coagulation and Thoroughfare-Conception vessel maladjustment types, 43 (81.1%), 24 (80.0%) and 29 (78.4%) were cured, 7 (13.2%), 5 (16.7%)and 5 (13.5%) markedly improved, 3 (5.7%), 1 (3.3%) and 2 (5.4%) improved respectively, and 1 (2.7%) of Thoroughfare-Conception vessel maladjustment type failed. A two-years’ follow-up showed that 3 of the cured 92 cases had a relapse, and after 2 more courses of treatment, they were cured again. Conclusion: Acupuncture therapy is effective for the treatment of hyperplasia of mammary glands.

Research Progress on Classical Prescriptions in Treating Mammary Glands Hyperplasia

According to Zhongjing Zhang's academic thought,gynecological diseases mostly result from deficiency,accumulated cold,and qi stagnation,so the treatment of gynecological miscellaneous diseases should take qi and blood as the general principle,and focus on the liver.The pathogenesis of mammary glands hyperplasia(MGH)is liver and kidney deficiency,disharmony of the thoroughfare vessel and the conception vessel,the causes of which are emotional imbalance,liver qi depression,and failure of transportation due to spleen deficiency.The superficial causes include qi stagnation,phlegm coagulation,and blood stasis,which are mainly related to the liver,spleen,kidney,the thoroughfare vessel,and the conception vessel.In clinical treatment of MGH,Xiaochaihu Decoction or Sini Powder can be used for qi stagnation syndrome,Xiao Xianxiong Decoction for phlegm coagulation syndrome,Guizhi Fuling Pill or Dahuang Zhechong Pill for spleen deficiency syndrome,and Wenjing Decoction for the deficiency-cold syndrome.

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.

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.

DETECTION OF HUMAN HERPESVIRUS 6(HHV-6) DNA IN SALIVARY GLANDS BY THE POLYMERASE CHAIN REACTION

To assess the presence of HHV-6-Specific DNA in human salivary glands, eighteen specimens of salivary gland tissue were investigated using the polymerase chain reaction. Eight of nine parotid glands, five of seven submandibular glands and one of two sublingual glands were found to have amplification of the HHV-6-specific sequence. The findings suggest that salivary gland tissue is one of the potential sites for HHV-6 persistence following primary infection and that saliva is a vehicle for transmission of the virus.

ELECTROACUPUNCTURE TREATMENT OF 30 CASES OF HYPERPLASIA OF MAMMARY GLANDS

In the present paper,30 cases of hyperplasia of mammary glands were treated withelectroacupuncture in different times following menstruation,showing satisfactory results.The totaleffective rate was up to 96.67％.In addition,the symptom of mastalgla abated abviously after treat-ment.

Comparative Expression Profiling of Lactogenic Hormone Receptor and It’s Signaling Molecules of Bovine Mammary Glands during lactation

Milk synthesis is known to be modulated by peptide hormones such as prolactin (PRL), growth hormone (GH), and insulin-like growth factor I (IGF-I). Previous studies suggested that PRL and IGF-I acted directly on mammary epithelial cells and were involved in lactation. Meanwhile, GH is thought to be indirectly involved in lactation by stimulating the secretion of IGF-I. It is controversial as growth hormone receptors (GHR) is expressed in the mammary epithelial cells. In order to clarify whether GH acted directly on mammary gland tissue, we investigated the prolactin receptors (PRLR), IGF-I receptors (IGF-IR), and GHR as well as the gene expression levels of the downstream signaling molecule for each receptor in the mammary gland tissue of Holstein cows during different stages of lactation. The results revealed that the mRNA expressions of PRLR and IGF-IR were highest during early lactation, and the mRNA expression of the GHR was highest during mid-lactation. We also found that the expression profiling of the signal transducer and activator of transcription 5 (STAT5) genes was similar to that of the GHR gene. On the other hand, the expression profiling of the PRLR gene was similar to that of the SHP2 gene. These results suggest that GH acts on the mammary glands directly, milk synthesis and secretion are chiefly stimulated in mid-lactation, and the timing of the action is different for PRL and IGF-I.

Pregnancies Resulting from Transgenic Embryos with Human Lactoferrin Gene Produced by Somatic Cell Nuclear Transfer

[Objective] The aim of this study is to understand the effects of donor cell type,embryo stage,number and transfer position on the efficiency of goat transgenic clone.[Method] Using somatic cell nuclear transfer technology,the single goat fetal fibroblasts(GFF)and mammary gland epithelial cells(GMGE)harboring human lactoferrin(hLF)gene were transferred to the enucleated oocyte.Reconstructed karyoplast-cytoplast couplets were fused,activated,and cultured in vitro.Embryos at 2-8 cell stage were transferred into oviduct of synchronized recipients,and blastocysts were transferred into uterine horn.[Result] The pregnancy rate was similar between GFF and GMGE(oviduct transfer:26.47% vs.20.00%),and between oviduct transfer and uterine horn transfer(26.47% vs.25.00%)for GFF group;pregnancy rate in the group with the mean number of embryo transferred per recipient of 21.2 was significantly higher than in those the 5.93 group and 9.64 group(40.00% vs.26.67% and 21.43%).[Conclusion] These results indicate that pregnancy rate of goat transgenic clone couldn't be affected by donor cell type,embryo stage and transfer position but be done by the number of embryo transferred per recipient.In addition,the study also suggests the feasibility of making transgenic goat using GMGE as donor cells.

Salivary gland antigens of laboratory-bred Phlebotomus sergenti and their immunogenicity in human volunteers in laboratory condition

Objective:To investigate Phlebotomus(P.)sergenti Parrot,1917(Diptera:Psychodidae)salivary gland antigens and their immune response in human.Methods:Human volunteers were exposed to sand flies’bites in the laboratory,and following each exposure the size of induration was recorded.The mean protein concentration of salivary gland lysate and specific anti-P.sergenti saliva IgG was measured.Sand fly salivary proteins were separated by SDS-PAGE and their immunoreactivity was examined by Western blotting assays.Results:Individuals exposed to P.sergenti salivary gland lysate for 8 months showed both antibody and delayed type hypersensitivity responses,although exposure for one month did not provoke any immune responses.The trend of antibody fluctuated during the exposure time and dropped by the end of antigen loading.The mean protein content was(0.36±0.08)μg in each pair salivary glands.Salivary gland lysate showed 11 to 12 major protein bands and 3 to 6 of them were immunoreactive.Conclusions:Our study showed that the salivary gland components of P.sergenti provoked both cellular and humoral immune responses in human.Furthermore,there are some immunogenic proteins in P.sergenti saliva which could be subjected for further investigation as vector-based vaccine candidate/s against anthroponotic cutaneous leishmaniasis.

Benign lymphoepithelial cyst of parotid gland without human immunodeficiency virus infection:A case report

BACKGROUND Benign lymphoepithelial cyst(BLEC)of the parotid gland is a rare benign embryonic-dysplastic cystic tumor in the anterolateral neck that occurs most commonly in human immunodeficiency virus(HIV)-positive adults and rarely in non-acquired immune deficiency syndrome patients.The main presentation is a slow-growing,painless mass,and secondary infection may cause acute inflammatory symptoms.CASE SUMMARY A 44-year-old Chinese male patient presented with a 1-year history of a mass in the left side of the neck.On physical examination,a mass similar in size and shape to a quail egg was found in the left parotid gland.The mass was tough,without tenderness,and easily moveable.The results of HIV tests,including antibody and nucleic acid tests and CD4+T cell examination,were negative.Imaging examination revealed a left parotid gland mass.The patient underwent surgical treatment,and BLEC was diagnosed based on postoperative pathology.After 2years of follow-up,the patient survived well without related discomfort.CONCLUSION The detailed characteristics of a BLEC in a patient without HIV infection contribute to an improved understanding of this rare disease.

Salivary gland disease in human immunodeficiency virus/acquired immunodeficiency syndrome: A review

The effect of human immunodeficiency virus(HIV) infection on salivary glands has diagnostic and prognostic significance. HIV-salivary gland disease(HIV-SGD) is comprehensively ascertained amongst the major critical acquired immunodeficiency syndrome(AIDS)-relatedoral manifestation and causes substantial morbidity. Parotid gland swelling due to sicca syndrome, parotid lipomatosis, sialadenitis, diffuse infiltrative lymphocytosis syndrome, benign lymphoepithelial lesions, neoplasms(benign or malignant) of salivary gland, parotid gland inflammation, diminished flow rates of saliva and xerostomia have been documented that also affects the health- associated characteristics of life in subjects infected with HIV. There is a necessity for health care researchers to diagnose it, particularly as it might worsen if left undiagnosed. The precise characteristic of alterations in dynamics of salivary gland structure and functionality with long-standing usage of highly active anti-retroviral therapy still remains unknown. HIV positive children also present with bilateral parotid enlargement and the syndrome state with classical clinical and cytological features of predominated lymphoid hyperplasia. Though various case reports and studies have been extensively published on different aspects of HIV-SGD, it has not been described solely, thus leading to occasional confusion of nomenclature and clinical presentation of HIV-SGD. This article reviews the pathogenesis of HIV-related SGD and its components and various other miscellaneous disorders affecting the salivary glands in HIV/AIDS.

Expression of biologically active human clotting factor Ⅸ(hFⅨ) in the mammary gland of transgenic mice

The DNA of human factor Ⅸ (hFⅨ) gene vector pMCⅨm, which had been proven to be able to express in in vitro and living cells, was introduced into 586 zygotes of Kunming White Mice by positive pressure microinjection technique with manual operation. The 499 survival embryos after microinjection were then transferred into pseudopregnant recipient mice and 216 F 0 pups were born. The analysis of PCR and Southern blot hybridization showed that, of the 216, 6 (2 females and 4 males) were integrated with foreign DNA in their genomes, giving an integration frequency of 3% (6/216). Two F\-0 female transgenic mice could express hFⅨ protein in their milk and the content was over 100 ng/mL as measured with ELISA. The biological activities of hFⅨ in the milk of two F\-0 mice were 44 67% and 79 43%, respectively.

Human clotting factor Ⅸ (hF Ⅸ ) secretion in goat milk after direct transfer with hF Ⅸ minigene into mammary gland by using retroviral vectors

THE mammary gland bioreactor system of lactating animal, established by germ-linemicroinjection and embryo transplantation, has been one of the most exciting projects in thefield of biotechnology. However, this system is costly, laborious and tim-cosuming. In 1994,Johanna et al. reported that secretion of a foreign protein, human growth hormone (hGH),in milk had been achieved after direct introduction of the cDNA into the mammary gland ofgoats by replication-defective retroviral vector. In that work, the in vivo expression of

Highly efficient expression of human extracellular superoxide dismutase(rhEcSOD)with ultraviolet-B-induced damage-resistance activity in transgenic silkworm cocoons

Extracellular superoxide dismutase(EcSOD)protects tissues from oxidative stress,and thus is considered as a therapeutic agent for many diseases such as atherosclerosis,hypertension,and cancer.However,cost-effective production of bioactive recombinant human EcSOD(rhEcSOD)remains a challenge.Herein,we developed an efficient strategy for producing active rhEcSOD by transgenic silkworms.rhEcSOD was successfully synthesized as homodimers and homotetramers in the middle silk gland and spun into the cocoons with a concentration of 9.48±0.21 mg/g.Purification of rhEcSOD from the cocoons could be conveniently achieved with a purity of 99.50%and a yield of 3.5±0.5 mg/g.Additionally,N-glycosylation at the only site of N89 in rhEcSOD with 10 types were identified.The purified rhEcSOD gained the potent enzymatic activity of 4162±293 U/mg after Cu/Zn ions incorporation.More importantly,rhEcSOD was capable of penetrating and accumulating in the nuclei of cells to maintain cell morphology and attenuate ultraviolet B-induced cell apoptosis by eliminating reactive oxygen species and inhibiting the C-Jun N-terminal kinase signaling pathway.These results demonstrated that the transgenic silkworm could successfully produce rhEcSOD with enzymatic and biological activities for biomedical applications.

Efficacy of Lubeikangru formulation in mice with hyperplasia of the mammary glands induced by estrogen and progesterone

OBJECTIVE: To evaluate the protective effects of Lubeikangru formula(LF) on hyperplasia of the mammary glands(HMG) induced by estrogen and progesterone in mice.METHODS: Female mice were divided randomly into five groups: normal, model, tamoxifen(3 mg/kg),Rupixiao(900 mg/kg) and LF(900 mg/kg). All mice except those in the normal group were treated sequentially with estradiol and progesterone to induce HMG. From the tenth day of induction, mice in normal and model groups received distilled water and mice in the other groups were given the corresponding drugs by gavage, once a day, for 30 d.At the end of treatment, the mammary glands, ovaries, hypothalamus, and serum was collected for whole-mount and hematoxylin and eosin(HE)staining, enzyme-linked immunosorbent assays(ELISAs), or western blotting.RESULTS: Whole-mount and HE staining of mammary glands showed that LF rescued(at least in part) the hyperplasic morphology of the mammary glands, and the number of branch points decreased after LF treatment(P < 0.05). ELISAs revealed that levels of estrogen and progesterone were decreased following LF treatment, whereas levels of gonadotropin-releasing hormone, follicle-stimulating hormone, and luteinizing hormone were increased in serum and tissues. Western blotting confirmed that LF treatment led to a reduction in expression of phosphorylated(p)-Erk, p-p38 and p-c-Jun N-terminal kinase. LF was also confirmed to be safe by acute-toxicity tests.CONCLUSION: LF can protect the mammary glands of mice from estrogen-and progesterone-induced hyperplasia by adjusting hormone levels and regulating the mitogen-activated protein kinase pathway.