Textual Research on Tibetan Medicinal Herb Lamiophlomis Herba / 中国实验方剂学杂志

Lamiophlomis Herba， one of the medicinal herbs commonly used by the Tibetan ethnic minority， has the effect of activating blood， stopping bleeding， dispelling wind， and relieving pain and is frequently used to treat trauma， traumatic bleeding， rheumatic arthralgia， and dampness and dampness-heat. By a review of ancient Tibetan medicine classics， materia medica classics， and modern literature， this paper systematically analyzed the name， nature， taste， original plant， medicinal part， harvest， processing， efficacy， and indications of Lamiophlomis Herba. The textual research showed that Lamiophlomis Herba was first recorded in the Somaratsa （《月王药诊》） in the middle of the eighth century. This medicinal herb was mainly recorded with a plain and warm nature and a sweet， bitter， and astringent taste. The herb was recorded as non-toxic in other books except the Tibetan Medicinal Plants in Gannan of Qinghai-Tibet Plateau （《青藏高原甘南藏药植物志》）. In the books of the past dynasties， the aboveground part of Lomiophlomis rotatd was used as medicine. In addition， there were also records of using the whole herbs of Ajuga ovalifolia， A. ovalifolia var. calantha， and Oreosolen wattii as the medicine. In ancient times， the herb was mainly harvested before Frost's Descent in the 8th-9th months of the lunar calendar， while it was mainly harvested during the flowering-fruiting stage in autumn in modern times. Due to the decreased reserve of Lamiophlomis Herba， the medicinal part evolved from whole herb or fresh leaves to the aboveground part. According to the ancient and modern records， this herb mainly has the functions of nourishing bone and marrow， eliminating dampness and dampness-heat， stopping bleeding， and relieving pain. In ancient times， it was used for treating parasitic diseases， menostaxis， spermatorrhea， diarrhea， and nourishing the body. This paper aims to provide a basis for further development and study of Lamiophlomis Herba through the textual research.

Effects of prenatal stress on learning and memory of the rat offspring / 西安交通大学学报(医学版)

Objective To determine the effect of chronic restraint stress during the late pregnancy on the behavior parameters of learning and memory of the rat offspring. Methods Stress was inflicted each day of pregnancy from day 13 to day 20. Pregnant females were individually restrained for 45 min three times a day. After delivery, one-month-old and three-month-old female and male rats were examined in Morris water maze to analyze the effect of exposure to prenatal restraint stress on the spatial learning and memory. Results In comparison to the control group, the latency to find the platform in the prenatally stressed group was significantly prolonged in both one-month-old and three-month-old offsprings (P

Effects of selenium and iodine on the expression of c-fos and c-jun mRNA and their proteins in cultured rat hippocampus neurons / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the effect of selenium (Se) and iodine (I) and the compound of both on the proto-oncogenes c-fos and c-jun mRNA and their protein expression in the cultured rat hippocampus neurons.</p><p><b>METHODS</b>Using the technique of serum free hippocampus neuron culture, different doses of Se and I and Se + I compound were added into the medium. The expression of the mRNA of c-fos, c-jun in hippocampus neurons cultured for 1, 3, 5, 7 and 10 d were studied using both in situ hybridization and SABC immunohistochemical technique.</p><p><b>RESULTS</b>Both Se and I could enhance the expression of c-fos, c-jun mRNA and their proteins, especially the combination of I and Se able to give a remarkable effect on c-jun mRNA expression.</p><p><b>CONCLUSIONS</b>Se and I may effect the expression of both c-fos and c-jun mRNA, especially the c-jun mRNA and its protein of hippocampus neurons, and thus may effect the differentiation and development of neurons.</p>

Studies on survival and outgrowth of processes of cultured rat hippocampus neurons in containing selenium and free serum medium / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effect of selenium on cultured newborn rat's hippocampus neurons survival and outgrowth development.</p><p><b>METHODS</b>Using the technique of primary culture of hippocampal neurons of newborn rat. The different dose of Se (62.5 microgram/L, 125.0 microgram/L, 182.5 microgram/L) were added into the medium at same time. We not only investigated the number of survival of neurons on 1 - 14 d and 1d to 10 d in with and without serum containing-Se medium, but also observed the length outgrowth of the neurite at 16 h, 24 h, 36 h, 48 h during culture.</p><p><b>RESULTS</b>Selenium could obviously enhance the outgrowth of early processed in 10% fetal serum medium and average length of neurite outgrowth is 15 - 20 micrometer more longer than control groups (P < 0.01) and selenium could also increase the livability of neurons and prolong survival time of cultured neurons in serum-free medium.</p><p><b>CONCLUSIONS</b>Selenium may play a very important role for early processed growth and development of hippocampal neurons.</p>

THE OVEREXPRESSION OF APOPTOSIS -RELATED GENES OF P53 AND BCL-2 IN CERVICAL CARCINOMA / 药物分析学报

Objective To investigate the significance of overexpression of P53 and bcl-2 protein in carcinogenesis of cervix. Methods 10 cases of cervical intraepithelial neoplasis(CIN) and 57 cases of invasive cancer were investigated with immunohistochemistry technique. Results The overexpresion of P53 protein in CIN and cervical cancer was significantly higher than that of control, respectively (P＜0.01). But there was no significant difference between CIN and cervical cancer(P＞0. 05). The immunoreactivity of bcl-2 in CIN was much more higher than that of control (P＜0.05). The positive rate and immunoreactivity of bcl-2 in cervical carcinoma were both remarkably higher than those of control (P＜0. 01) ,but there was no significant difference between CIN and cervical carcinoma (P＞0. 05). It was also found that there was a remarkably positive correlation between the overexpression of bcl-2 and P53 (P＜0.01). Conclusion Because of the loss of wtP53 function,the expression of bcl-2 can not be down-reguated,which is associated with the pathogenesis and development of cervical carcinoma.

CLONING AND EXPRESSION OF A GENE MEDIATINGγ-RADIATION-INDUCED APOPTOSIS IN HL-60 CELLS / 药物分析学报

Objective To identify the member of the caspase family proteases involved in γ-radiation-induced apoptosis in HL-60 cells and to study the expression of the caspase gene in normal, apoptotic cells and in immortal tu mor cells. Methods By using degenerate oligonucleotide primers encoding the highly conserved peptides that were pre sent in all known caspases, we performed RT-PCR on poly(A)RNA from γ-radiation-induced apoptotic HL-60 cells. Caspase-3 mRNA in apoptotic HL-60 cells and in human tumor cell lines was analyzed by Northern blot. Results The amplified DNA fragment was identified with caspase-3 cDNA by cloning and sequencing. The Northern blot analysis of caspase-3 mRNA of different human tumor cell lines showed that the caspase-3 gene transcript was more highly ex pressed in leukemia cell lines and the SH-SY5Y cell line than in HeLa and MCF-7 cells. It was more highly expressed in the radiation-induced apoptotic HL-60 cells than in control HL-60 cells. Conclusion These results indicated that caspase-3 was involved in γ-radiation-induced apoptosis in HL-60 cells. The high level of expression of caspase-3 may aid efforts to understand the insensitivity of some tumor cells to radiation, their inherent ability to survive, and apop tosis.

ANATOMICAL RELATIONSHIPS OF SEROTONINERGIC,DOPAMINERGIC,HYDROXYLASE,NEUROPHYSINERGIC AND ENDORPHINERGIC PROJECTIONS WITH THE GNRH SYSTEM IN THE DIAGONAL BAND OF THE RAT / 解剖学报

The distribution relationships of gonadotropin-releasing hormone (GnRH) (5-HT), tyrosine hydroxylase (TH), neurophysin (NP) and ?-endorphin (?-END) in the diagonal band of Broca (DBB) of rat brain were studied by immunohis- tochemistry. The results indicate that there was a close spatial overlap in the distribution between 5HT,TH,NP,?-END fibers and GnRH perikarya and fibers in the ventromedial area of VDBB and dorsomedial area of HDBB. In the dorsomedial area of VDBB close overlap in the distribution between 5-HT fibers and GnRH neurons was also observed. In addition, some TH and NP immunoreactive neurons were found in the medial areas of VDBB and HDBB, The close relationships in distribution and functional significance of GnRH,5-HT, TH, NP and ?-END were discussed.

IMMUNOHISTOCHEMICAL STUDY OF POSTNATAL DEVELOPMENT OF GNRH NEURONS IN THE RAT BRAIN / 解剖学报

The morphology of the GnRH neurons in the rat brain during postnatal development was studied quantitatively using the ABC immunostaining method and an image analyser. The pattern of distribution of the GnRH neurons in different ages animals used was similar. GnRH cells of rats aged 1-7 days were small in size with short processes, pale-stained, and smooth in outline The number of GnRH cells in 1 day rats was not significantly different from that of adult, but a significant decrease in the GnRH cell number appeared in 7 day rats. In postnatal day 14, Gn-RH neurons reached adult level in cell body size, number and staining intensity. The GnRH fibers and terminals in the median eminence were gradually increased and reached adult level up to 14 postnatal days. These results suggested that the number of GnRH neurons is determined shortly after birth; the second postnatal week is the critical period for the development of GnRH neuronal morphology; and the establishment of function of the GnRH neuronal system may be earlier than its morphological maturation.

Effect of prenatal restraint stress on the expressions of Bax, Bcl-2, Fas and FasL in hippocampus of the offspring / 西安交通大学学报(医学版)

Objective To study the effect of chronic restraint stress on the expressions of Bax, Bcl-2, Fas and FasL in hippocampus of the offsprings during the pregnancy. Methods Restraint stress was performed three times each day of pregnant women from day 13 to day 19 (each time for 45min). After delivery, the expression of Bax, Bcl-2, Fas and FasL in hippocampus of one-month-old female and male rats were respectively examined by immunohistochemical method. Results Compared with the control group, the expression level of Bax in stressed group is higher (P 0.05). Conclusion Restraint stress during the pregnancy can upgrade the expression of Bax and degrade the expression of Bcl-2 of the offsprings; no effect was found on the expressions of Fas and FasL of the offsprings.

GNRH NEURONS AND THEIR PATHMAYS IN THE BRAINS OF RATS AND MICE——AN IMMUNOHISTOCHEMICAL STUDY / 西安交通大学学报(医学版)

By means of ABC immunoenzyme technique, the distribution of GnRH neurons and fibers in the brains of rats and mice was observed, The results showed that more than 90% of GnRH neuronal perikarya were concentrated in the diagonal band-medial preoptic area near the organum vasculosum lamina terminalis, extending rostro-dorsally to the medial septic nucleus and caudo-laterally to the supraoptic nucleus region. No GnRH cell body could be found in the mediobasal hypothalamus, GnRH fibers terminated in the median eminence via the periventricular and lateral tracts. In the rostral median eminence, fibers were present in its entire layers and width, and caudally separated into two laterally-located bundles. The difference of GnRH system between rodents and primates, as well as its significance in reproduction are discussed.

CYTOLOGY OF MAGNOCELLULAR NEUROSECRETORY NEURON OF HYPOTHALAMUS IN RATS / 西安交通大学学报(医学版)

By means of chromalum-hematoxylin and thiosulfate aldehydefuchsin stains the magnocellular neurosecretory neuron of the rat hypothalamus was studied. The cell body of neuron containing various amounts of secretory granules was large in size, with a variety of shapes. The axon of neuron showed an beaded appearance; only one in each neuron was very fine in diameter and left hypothalamic nuclei to form the hypothalamo-neurohypophyseal tract. The dendrite, in general, thicker than the axon, did not project outside of the nucleus. Many of the thick short dendrites did not contain secretory granules or only few secretory granules. The thick processes with more secretory granules were considered as dendritic processes. A number of axons and thick processes containing secretory granules also contacted with the endothelium of vessels and the ependyma of the third ventricle. In addition, a group of magnocellular secretory neurons were found in the sub-choroid plexuses of the lateral ventricle.

ABNORMAL EXPRESSION OF TYROSINE HYDROXYLASE IN THE CEREBELLUM OF NIEMANN-PICK TYPE C MICE / 解剖学报

Objective To investigate the time, location and relationship to clinical manifestation of abnormal expression of tyrosine hydroxylase(TH) in the cerebellum of Niemann\|Pick type C disease (NPC). Methods Immunohistochemical staining was applied, by using antibodies to TH, to brain sections from NPC and normal mice of various ages. Some adjacent sections were stained for calbindin D28k. Results There was no marked decrease in number of cerebellar Purkinje cells(PC), which were negatively stained for TH, in the NPC mice aged 1\|3 weeks. The PC were decreased in number starting from week 5. During postnatal weeks 8\|11, PC were significantly lost, but considerable number of PC in the nodulus and uvula vermis survived. Some survived PC and their dendritic trees were TH immunoreactive after week 8. These PC showed axonal spheroids and irregular dendrites that were bent, broken, locally enlarged or atrophied. Conclusion Mutation of NPC1 gene induces severe loss of cerebellar PC and survived PC have been damaged morphologically and show abnormal gene activity. These may be the pathogenic basis of movement disorders of NPC.

EFFECTS OF GOSSYPOL ON THE ULTRASTRUCTURE OF THE CELLS OF RAT ADENOHYPOPHYSIS / 解剖学报

The ultrastruetural changes of the cells of adenohypophysis were observed in adult male rats following administration of gossypol at a daily dosage of 30 mg/kg for 5 weeks. The results indicated a remarkable increase in the secretory activity of gonadotrophs. In comparison with the control group, the number of the less actively functioning types Ⅰ and Ⅱ gonadotrophs of gossypol-treated rats decreased, while that of types Ⅲ and Ⅳ cells with developed Golgi complex and dilated rough endoplasmic reticulum increased. Moreover, castration-like cells (type Ⅴ) and degranulation cells (type Ⅵ) appeared in the experimental animals. No significant changes in the other tropic cells of adenohypophysis could be found. The classification of gonadotrophs and the possible mechanism of gossypol-induced changes in them are discussed.

IMMUNOCYTOCHEMICAL STUDY OF GONADOTROPIN-RELEASING HORMONE (GnRH) NEURONS IN DISSOCIATED CULTURES OF NEWBORN RAT HYPOTHALAMUS / 解剖学报

Gonadotropin-Releasing Hormone (GnRH) immunoreactive neurons in dissociated cell culture from newborn rat hypothalamus were investigated on days 1,3,5,and 7 in vitro by means of the im-munocytochemical method. The results showed that GnRH was expressed in the first day of culture. GnRH neurons accounted for 12.1-14. 8% of the total neurons in culture,and they were mainly bipolar in type. There were growth cones on the end of GnRH processes. Various patterns of intercellular contacts between GnRH neurons and between GnRH and other neurons were also observed. These findings indicate that the cultured GnRH neurons exhibit the morphological and functional characteristics of the GnRH neurons in vivo,and serve as morphological evidence for pulsatile secretion of GnRH and its regulation.

ULTRASTRUCTURAL OBSERVATIONS OF THE GOSSYPOL EFFECT ON THE RAT HYPOTHALAMUS / 解剖学报

Observations of the ultrastructural changes in neurons of hypothalamie arcuate, supraoptic and paraventricular nuclei were carried out in the rats following gossypol treatment at a dosage of 30mg/kg/day for 5 weeks. The results showed that after gossypol treatment no marked changes in supraoptic and paraventricular cells could be seen at electron microscopic level, and that there appeared a series of ultrastrucrural changes reflecting increased neurosecretory activity in some of arcuate neurons.They were as follows: Golgi complex and rough endoplasmic reticulum were slightlydilated and filled with homogeneous, low electron density materials in their cisternae; the amounts of neurosecretory granules, lysosomes, nucleolus-like bodies, and microtubules were increased. The nature, significance and cause of these changes are discussed.

ISOLATION,CULTURE AND VERIFICATION OF STROMAL AND GLANDULAR CELLS OF HUMAN ENDOMETRIUM / 解剖学报

Objective Stromal and glandular cells of human endometrium were successfully isolated and cultured in this experiment. Methods These two types of cells were observed by light microscopy and verified by immunocytochemical staining. Results The result showed that some of stromal cells became adherent after.0 5h of culture.Two shapes of stromal cells were found.One was spindle and the another was polygonal.Immunocytochemical staining method showed that both shapes of stromal cells were positive for vimentin,with 95% of positive rate,and negative for cytokeratin 1,indicating that these cells were endometrial stromal cells.Most of glandular cells became adherent after 24h of culture and formed tightly packed whorls after 4d.Individual cells were polygonal and had a large and round nucleus.Immunocytochemical staining method showed that glandular epithelial cells were positive for cytokeratin 1,with 90% of positive rate,suggesting their nature of epithelial cells. Conclusion Stromal and glandular cells of human endometrium were successfully isolated by using two series of filter and cultured in this experiment. [

IMMUNOHISTOCHEMICAL STUDY ON DISTRIBUTION OF Gn RH-ASSOCIATED PEPTIDE IN THE FOREBRAIN AND HYPOTHALAMUS OF RODENTS / 解剖学报

Gonadotropin-releasing hormone(GnRH)prohormone of human and rat consistsof GnRH and GnRH-associated peptide(GAP).In this study,three antisera againstN-terminal,mid-region and C-terminal of GAP,and ABC immunoenzyme methodwere used to observe the GAP neurons in the brains of the rat,mouse and guineapig.The distribution of GAP neurons in these animals was similar.GAP perikaryawere mainly present in the septo-preoptic area,with the largest concentration inthe diagonal band near the organum vasculosum of the lamina terminalis.SomeGAP perikarya were also seen in the brain area near the supraoptic nucleus.GAPfibers were widely present in the forebrain and hypothalamus,and terminated inthe organum vasculosum of lamina terminalis and median eminence.Of 3 GAP antisera,the one against N-terminal gave more immunoreactive elements and moreintense staining.The morphology and distribution of GAP perikarya,fibers andterminals were similar to those of GnRH.These results,combined with other relatedfacts,suggest that there is a common GnRH prohormone in mammals,and itsprocessing products,GAP(or cleavage fragments)and GnRH,are cosecreted intohypophysial portal system to regulate anterior pituitary hormone secretion.

EFFECT OF Caspase-3 mRNA ANTISENSE OLIGODEOXYNUCLEOTIDES ON ?-IRRADIATION-INDUCED APOPTOSIS OF HL-60 CELLS / 解剖学报

Objective To observe the inhibitory effect of caspase\|3 mRNA antisense oligodeoxynucleotides (ASODN) on the apoptosis of HL\|60 cells, and to screen the effective ASOND. Methods By means of the lipofectimine\|mediating transfection, caspase\|3 mRNA ASODNs of four different sequences were introduced into HL\|60 cells followed by ? irradiation.The gel electrophoresis was performed for the DNA ladder, Hoechst 33258\|propidium iodide fluorecent staining for the percentage of apoptotic cells, and FCM for quantitative analysis of apoptosis. Results When HL\|60 cells were transfected with caspase\|3 mRNA ASODNs targeting 5'noncoding region (sites -62 to -46) and initiative coding region (sites -1 to 16) at the final concentration of ≥3?? mol/L, the DNA ladder was not detected with the gel electrophoresis, nor was the apoptotic peak found with FCM. Fluorescent staining analysis showed that the percentage of apoptotic cells was significantly lower than that in the control groups including non\|transfection group and mismatched oligodeoxynuleotide group ( P

HISTOLOGICAL AND HISTOCHEMICAL OBSERVATION OF THE GOSSYPOL EFFECT ON THE SUPRAOPTIC,PARAVENTRICULAR AND ARCUATE NUCLEI IN RAT HYPOTHALAMUS / 解剖学报

In order to analyze whether gossypol has an effect or what effect it has on the hypothalamus, the general structure and morphologic indicators reflecting the neurosecretory function of the neurons of hypothalamic supraoptic, paraventricular and arcuate nuclei were observed in adult male rats following gossypol administration (30 mg/kg/day ? 5 weeks, and 30 mg/kg/day ? 5weeks + 60 mg/kg/week ? 5weeks). Results showed that no marked change in the light-microscopic morphology of the neurosecretory cells of the above mentioned nuclei was found after gossypol treatment for 5 or 10 weeks; and that no significant change appeared in the neurosecretory activity of the three nuclei, according to nuclear volume, contents of Nissl bodies and neurosecretory materials, and activity of thiamine pyrophosphatase and acid phosphatase, merely the Nissl body content in paraventricular nucleus increased after feeding gossypol for 5 weeks. The data indicated that the effective antifertility dose of gossypol had no direct influence on the hypothalamus of adult male rats. It was possible that the temporary increace in the secretory activity of paraventricular nucleus reflected physiologically regulating response of the magnocellular neurosecretory system to some change in the internal milieu of organism caused by gossypol.

EFFECTS OF ETHANE DIMETHANESULPHONATE ON THE HYPOTHALAMO-PITUITARY-TESTICULAR AXIS OF THE RAT / 解剖学报

By means of histological and immunohistochemical methods,effects of a singleintraperitoneal injection of ethane dimethanesulphonate(EDS,75 mg/kg bodyweight)on the hypothalamo-pituitary-testicular axis of the rat were observed inthis study.Three days after administration of EDS,Leydig cells were eliminated;some LH cells in pituitary became larger or decreased in immunoreactivity;but nosignificant change could be found in hypothalamic GnRH system.Seven days afterEDS,degeneration of spermatogenic epithelium was marked,the staining intensity ofLH cells was generally reduced,the number of GnRH immunorcactive neurons inthe hypothalamus as well as the density and staining intensity of GnRH fibers andterminals in the median eminence were significantly diminished.Testosteronereplacement increased the number of late spcrmatids in testis and restored theabove-mentioned changes in the hypothalamus and pituitary.These results indicate that following administration of EDS,1)spermatogenesisdamage is resulted from destruction of Leydig cells and cessation of testosterone secretion;2)respective hormone release from hypothalamic GnRH system andpituitary LH cells is increased;3)loss of testosterone negative feedback is themajor factor responsible for the enhancement of secretory activity of the hypotha-lamo-pituitary axis;and that 4)EDS can be a useful experimental tool forstudying hypothalamo-pituitary-testiculer axis and intratesticular local regulation.