Objective:To study the change rule of decocting quantity of the effective components in Amomi fructus and Amomi fructus rotundus with decocting time to determine whether or not decocted later and optimal decocting time. Methods:The herbs were extracted by the traditional water decoction, and at different time points, sampling was carried out. Using camphor and eucalyptol as the index components, the change rule of decocting quantity of the effective components with the decoction time under the condition of single and combined decoction was investigated. Results:When the decoction time of Amomi fructus was within the range of 3-6 min, the total amount of camphor in the decoction reached relatively high value, and the total amount lost more than 45% when the decoction time exceeded 10 min. Amomi fructus rotundus boiled for a short time below 2 min, and when the decoction time was more than 5 min, more than 50% eucalyptol lost. Conclusion:Amomi fructus and Amomi fructus rotundus should be decocted later with decocting time within 3-6min and below 2 min, respectively. The analytical method is reliable and precise in the quality control of relative decoction.

A series of monoclonal antibodies have been used to study the distribution of T and B lymphocytes and their subsets in human spleen (5 from normal human and 2 from patients with portal hypertension). The results indicate that the T cells are mostly located in the periarterial lymphatic sheath, and in which a few B lymphocytes can be seen. The B cells are concentrated in lymphoid follicles, but also contain some T lymphocytes such as Leu 1, Leu 3a and Leu 4 positive T cells, these cells are necessary for forming of germinal center. Whereas the marginal zone, is composed of a mixture of T and B cells as well as the T_(ac) positive cells. The red pulp is composed of a mixture of T and B ceils, but the T and B cells are distributed randomly. In this report, the LN-2 monoclonal antibody is used first to study the B lymphocytes in human spleen. So far it is a unique antibody to react with nuclear membrane of B lymphocytes, the activity of LN-2 antigen do not influenced by B-5 fixation and paraffin embedding. From our data, there is no difference in staining feature and charateristic distribution between the normal human spleen and spleen of portal hypertensive patients. Although the periarterial lymphatic sheath in cases with portal hypertension seems to be narrower than the normal.

The mucous glands (sublingual gland, pyloric gland, main pancreatic duct gland, duodenum gland and large intestinal gland) have been studied-by means of labelled lectins and histochemical methods. The results indicated that a large amount of mucopolysaccharide with sailic acid are found in sublingual glands and pyloric glands, while a large amount of sulfate mucopolysaccharide are observed in the other three glands. In addition, the reactivity of DBA and PNA in mucous glands are different, for example, the sublingual and large intestinal glands are negative for DBA and PNA staining. On the contrary, the pyloric glands, duodenum glands as well as main pancreatic duct glands are heavily stained with DBA, but various between the mucous glands stained with PNA. The goblet cells in duodenum, large intestine and main pancreatic duct are similar in their morphology, but the reactions of PAS, AB and lectins are different between them, even so in the same organ, the staining pattern also differ from their locations. The mucous glands and goblet cells contained different mucopolysaccharides, which might be concerr/ed with different functions in different organs.

Leu 7 monoclonal antibody and immunohistochemical technic were used to study the distribution of NK cells in human lymph node, tonsil, spleen and thymus. The results indicated that the NK cells predominately distributed in B cell area, such as germinal center of secondary follicle, which occured in lymph node and tonsil. A few NK cells were found in the paracortex and medulla of lymph node. In the spleen, the NK cells mainly located in the germinal center of splenic nodules and in the periphery of white pulp. However, the NK cells were never shown in the perarterial lymphatic sheath, some NK cells also scattered in the pulp cord and sinuses of red pulp. In the thymus, the NK cells were restricted in the medulla, and the number of NK cells reduced in the aged thymus. In addition, the Leu 7 antibody was reacted with epithelial cells, which located in the cortical periphery or thymic lobule. The morphology of NK cells in different lymphoid organs was similar. The staining intensity of Leu 7 antibody was identical both in the frozen and paraffin sections.

9 fetal and 7 juvenile thymuses were used to identify the asteroid cells on the frozen sections by immunohistochemical method. The results indicated that the thymic medulla, except B cells, contains a type of larger cells with small cytoplasmic processes, which has in common with B cell antigens, such as 1gM, 1gA, Leu14, BA-1, HLA-DR and light chain-Kappa, but they do not express OKB-2. Moreover, the asteroid cells do not reactive with T cell antibody and follicular dendritic cell antibody, and lack of ANAE activity. The asteroid cells in the fetal thymus are constantly located in the corticomedullary region, while, in the juveniles, the asteroid cells are mainly distributed within medulla or in the vicinity of Hassall's corpuscles. As regards the function of asteroid cells is not clear. Based on their distribution, it is probably associated with T cell maturation. On the morphologic ground, the asteroid cells are similar to interdigitating cells in thymus, the relationship between them has been discussed.

Using immunoenzymatic technique,we have observed the localization ofimmunoglobulins in human palatine tonsils,of patients suffering from chronictonsilitis and hyperplasia.The result as follows:1.Epithelium:The outer surface of tonsil is lined by stratified squamous epithelium,the epithelium rarely shows keratinization and lacks a well-defind and continuousbasement membrane.In the basal layer of epithelium,it is reticular structure andspaces between the cords of epithelium cells are filled with Ig~+ lymphocytes andplasma cells,the reticular structure is particularly pronounced near the crypts.Thisspecial structure is regarded as an expression of active immune function.In thesuperficial layer of epithelium,the predominate cells are Ig~- small lymphocytes,maybe T lymphocytes.2.Follicle:All follicles have germinal centres,especially hyperplastic tonsils.The follicle has polarity,each follicle may be divided into three Zones(a b c),twozones(a b)form the centre of the follicle and the thired(c)is the mass of smalllymphocytes“capping”and enclosing the centre.The upper limits of zone(c)areoften indistinct,from the presence of many lymphocytes in the epithelium.Thedistribution of Ig~+ cells in follicle are different from their three zones.In the centre,IgG cells are most numerous,next most numerous are IgA~+ cells,and considerablyfewer IgM positive cells.In the c zone,only a few Ig~+ cells are seen(IgG IgM).Itis found that the germinal centre may produce more than one type of Ig,it differsfrom the report of Sordat,but quite agrees with Curran's result.However the numbersof cells containing the various types of Ig show a great difference.3.Other lymphoid tissue:The tissue between the follicles are rich in small Tlymphocytes and a few Ig~+ containing cells,this area was called thymic dependentarea.At an older age,this region is always enlarged,but the density of the cellswill decrease,meanwhile much connective tissues are seen in the stroma.

A sensitive immunocytochemical technique has been used to study the effect of Cordyceps Sinesis on the IgM and IgG antibody-forming cells in mouse spleen. The results indicate that after the mice have been treated for 7 days with Cordyceps Sinesis, their splenic nodules were increasing, the germinal centers became prominent and the marginal zones were thickening. The immunocytochemical results disclosed that a huge amount of IgM and IgG lymphocytes appeared in the splenic nodules, especially in the marginal zones, which were thickening due to the proliferation of the IgM-lymphocytes. In the red pulp, a large amount of IgM- and IgG-plasma cells were peripherally located in the terminal arteries or sinuses, indicating that it may be convenient for the plasma cells to release the antibodies into blood circulation. The results suggest that the Cordyceps Sinesis could stimulate the IgM-and IgG-lymphocytts to proliferate and promote the forming of antibodies.

BACKGROUND:Studies confirm that electromagnetic field (EMF) can promote the synthesis and secretion of many bone growth factors,and some growth factors can induce the osteoblastic directional differentiation of bone marrow mesenchymal stem cells (MSCs).OBJECTIVE: To investigate the effect of power-frequency EMF on mRNA expressions of bone morphogenetic protein-2 (BMP-2) and transforming growth factor-beta 1 (TGF-β1) in mouse bone marrow MSCs cultured in vitro.DESTGN: Single sample, block design, observation and controlled animal trial.SETTING: Department of Traumatic Surgery, Tongji Hospital Affiliated to Tongji Medical .College, Huazhong University of Science and Technology.MATERIALS: This trial was carried out in the laboratory of Department of Traumatic Surgery, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology during February to December 2005. ①Twenty Kunming mice of clean grade were selected for harvest of bone marrow MSCs. ②Magnetic field generator,which could generate EMF with 0 to 100 mT field strength and successive adjustable 50 Hz sinusoidal wave, was developed by Wuhan Naval University of Engineering. ③ Primer was all synthesized by Saibaisheng Bioengineering Co.,Ltd., Beijing.NETHODS: ① The involved mice were sacrificed by cervical dislocation. Bilateral femora and tibia were harvested. Bone marrow MSCs were isolated and cultured in vitro, and the second generation of cells were used for the trial. ②Different intensities of EMF stimulation tests: Negative control group, positive control group, EMF 0.4, 0.8 and 1.6 mT stimulation groups were set. Five bottles of cells of the second generation were chosen from each group for test. The cells in the negative control group and positive control group were not exposed to EMF. But medium containing osteogenic inductor(10-8 mol/L dexamethasone, 10 mmol/L β-sodium glycerophosphate and 50 mg/L Vitamin C included) was added in the positive control group at passage. After adhering to the wall, the cells in the EMF 0.4,0.8 and 1.6 mT stimulation groups were exposed to EMF of 0.4,0.8 and 1.6 mT field strength, respectively, one hour per day, five days later, they were detected.③ EMF stimulation tests in the same field strength and different time: Negative control group, EMF 1.6 mT stimulation 15,30 and 60 minutes groups were set.Five bottles of cells of the second generation were chosen from each group for test. The cells in the negative control group were exposed to EMF. The cells in the EMF 1.6 mT stimulation 15,30 and 60 minutes groups were respectively given 15,30 and 60 minutes of EMF stimulation at 1.6 mT successively.Five days later, they were detected.④ The mRNA expressions of BMP-2 and TGF-β1 were detected in each group by reverse-transcriptase polymerase chain reaction (RT-PCR) technique.MAIN OUTCOME MEASURES: ① The effect of different field strength of exposure of 50 Hz EMF on mRNA expressions of BMP-2 and TGF-β1. ② The effect of the same field strength and different time of exposure of 50 Hz EMF on mRNA expressions of BMP-2 and TGF-β1.RESULTS: ①Five days after EMF stimulation, the mRNA expressions of BMP-2 and TGF-β1 in the positive control group and EMF 0.4,0.8 and 1.6 mT stimulation groups were significantly enhanced as compared with negative control group (all P ＜ 0.01), and the mRNA expression of BMP-2 in the EMF 1.6 mT stimulation group reached the peak [(57.74±0.23)％]and mRNA expression of TGF-β1 in the EMF 0.4 mT stimulation group also reached the peak [(126.20±0.21 )％].② Five days after EMF stimulation, the mRNA expressions of BMP-2 and TGF-β1 in the EMF 1.6 mT stimulation 15,30 and 60minutes groups were significantly enhanced (all P ＜ 0.01) as compared with negative control group, and the mRNA expressions of two factors in the EMF 1.6 mT stimulation 60 minutes group reached peak separately [(28.06±0.11 )％ and (75.20±0.16)％].CONCLUSION:Proper intensity and action time of exposure of power-frequency EMF can obviously promote the mRNA expressions of BMP-2 and TGF-β1 in mouse bone marrow MSCs cultured in vitro.

Objective To develop an instrument that detects the endurance of an animal hanging on a bar during medical and pharmaceutical scientific experiments .Methods One pair of opto sensors were used for signal conversion .A solenoid valve device was used for water level .A 51-Series microcontroller was used to control the experimental setup and record the results .Results We have developed a microcontroller-based experimental instrument that can automatically detects the endurance of an animal hanging on a bar during medical and pharmaceutical scientific experiments .Our experimental results with 28 mice indicated that the applicability of the instrument is good .Conclusion The instrument provides an innovative tool for detecting the endurance of an animal .

The morphology and distribution of pre-B cells and the changes of antigens during the differentiation from pre-B to B cells in fetal livers of different gestational ages were studied by means of immunohistochemical technique. The results revealed that at earlyfetal stage(between 9 and 29 weeks), the liver contained numerous pre-B cells, which were different in shapes and sizes, but their phenotypic expression was identical, for instance, 1gM, BA-1, HLA-DR and TdT all appeared positive. Most of pre-B cells were scattered in perisinusoidal space, only a few of them were present in sinusoids or around the blood vessels. After the 13the week, 1gD and 1gA positive cells began to appear, and the number of OKB-2 and Leul4 positive cells was markedly increased subsquently. Meanwhile, the number of the HLA-DR, Kappa and Lambda positive cells was increased accordingly, which indicated that the B cells became more mature. In view of absence of antigenic stimulation, the B cells failed to develop into plasma cells. Moreover, it was observed that the differentiation and maturation of the B cells in the fetal liver seemed to be independent on the T cells, because the B cells continued to generate even if the T cells were absent at early stage of fetal liver. It showed that the differentiation and the development of the B cells in the fetuses were mainly dependent on the liver microenvironment. This study provided an evidence for treatment of aplastic anemia and agammaglobulinemia with fetal liver between 9 and 20 weeks old.