Objective To construct a subtractive cDNA library of genes differentially expressed in human lymphoma cell line Jurkat cells treated with glycyrrhizin (GL), and to clone genes associated with its immunological regulation, and to further elucidate the molecular immune mechanism of GL. Methods The mRNA was isolated from Jurkat cells treated with either GL or 0.9 percent sodium chloride as a control, then cDNA was synthesized. Suppression subtractive hybridization (SSH) method was employed to analyze the differentially expressed DNA sequence between the two groups. The cDNA was sequenced and analyzed in GenBank with Blast search after the amplification of the subtractive library by PCR. Results The amplified library contained 28 positive clones. Colony PCR analysis showed that there were 22 clones containing 200-1 000 bp inserts. Sequence analysis was performed, and the full length sequences were obtained with bioinformatics method. Altogether 11 kinds of encoding sequences were achieved, including interleukin-12, interleukin-18, and thymosin ?1, etc. Conclusions A subtractive cDNA library of genes differentially expressed in Jurkat cells treated with GL using SSH technique was constructed successfully, and it might give some new clues for the study of the immune regulation mechanism of GL.

Objectives: To reveal the morphological features of the stratum corneum lipids in hairless mouse epidermis. Methods: Ruthenium tetroxide and osmium tetroxide were compared as post fixative in the preparation of hairless epidermis for transmission electron microscopic examination. Results: Both reagents reveled characteristic membrane coating granules within the granular layer. Whereas, the transformation of the membrane coating granule contents into multiple lamellae at the interface between the granular and cornified layers and the persistence of these lamellae through all levels of the stratum corneum were demonstrated only by ruthenium tetroxide fixation. Conclusions: The distinctive patterning of the intercellular lamellae reflects the nonrandom organization of the stratum corneum lipids. In addition, the ruthenium tetroxide postfixation technique is a useful method in the investigation of the morphological features of the stratum corneum lipids.

Objective To explore the characteristics and treatment of pain after inflammatory spinal demyelination. Methods 271 patientssuffered from inflammatory spinal demyelination with pain were analyzed retrospectively. Results Acute radicular pain and Lhermitte'ssign were common in the acute pain syndromes. Individual therapy showed a benefit of decreased pain. Conclusion Pain is a commonclinical symptom of inflammatory spinal demyelination. Individualized therapeutic decisions could relieve symptom and improve outcome.

Objective To observe the therapeutic effects of nerve blockade combined with millimeter wave on patients with head acute herpes zoster (HZ) following serious pain.MethodsForty-two patients with head acute HZ following serious pain were randomly assigned to two groups: the combined group which treated with nerve blockade combined with millimeter wave, and the control group which received only nerve blockade. The pain degree (Numeric Rating Scales, NRS), blisters and scabbing were evaluated after treatment.ResultsNRS significantly decreased in both groups after treatment. The blister was earlier to disappear and scabbing was better in the combined group than in the control group (P<0.01).ConclusionNerve blockade combined with millimeter wave can effectively relieve the pain and shorten course of head acute HZ following serious pain.

Objective To investigate the incidence, possible mechanisms, clinical features of severe headache after posterior fossa craniotomy. Methods 119 neurosurgical patients undergoing selective posterior fossa craniotomy were analyzed retrospectively. Results 45 （37.8%） patients experienced severe postoperative pain, which could be identified as neuralgia (19 cases), cervicogenic headache (9 cases) and incision of scalp pain （17 cases）. Conclusion In addition to neuralgia and incision of scalp pain, cervicogenic factor plays an important role in severe headache after posterior fossa craniotomy.

Objective:To explore the effect of Leptin on the apoptosis of thymocytes induced by lipopolysaccharide(LPS) in mice.Methods:The toxicity of LPS on thymocytes was evaluated using MTT colorimetric assay.Thymocytes apoptosis was detected using Annexin V-FITC/PI double staining flow cytometry.caspase3 mRNA were measured by RT-PCR.Results:After cultured for 6 h,apoptosis of thymocytes was identified by flow cytometry.Apoptosis rate in LPS group (55.12?0.656)% was significantly higher than that in control group (9.58?0.605)% (P

Reproductive toxicity research takes an important place in traditional Chinese medicine pre-clinical safety evaluation. Modern reproductive toxicity experiment includes drug-related miscarriage, fetal death, teratism, and adverse effects on fertility, genital system, embryonic development and fetus, which is different from contraindicated in pregnancy in traditional Chinese medicine theory. Now the three-phases reproductive toxicity study is the method mainly applied in traditional Chinese medicine reproductive toxicity evaluation. Besides that, alternative methods of whole embryos culture and embryonic stem cell test are also used in traditional Chinese medicine embryo toxicity evaluation. This article reviews research progress and pre-clinical evaluation on reproductive toxicity of traditional Chinese medicine.

Objective　To study the histopathologic features and pathogenesis of mucosa-associated lymphoid tissue lymphoma (MALT-oma) of salivary glands. Methods　Clinical data, paraffin-embedded sections, immunohistochemical slides (SP method) and electron microscopic features of surgical specimens of 32 cases of salivary gland MALT-oma were studied. Results　The patients were 27 males and 5 females, with a mean age of 54.76 years. The lesions were located in the parotid area in 17 cases, and in the submandibular gland in the remaining 15 cases. Much of the MALT-oma was replaced by infiltration of a great amount of centrocyte-like cells (CCL) as background and occasional large cells (centroblast- or immunoblast-like). In MALT-omas “lymphoepithelial lesions" were present. Immunohistochemically, CD20 expression was found to be positive and CD45RO expression was negative in all MALT-omas. Conclusion　Most of the MALT-omas are low grade malignant tumors and have a “homing back" phenomenon. The cases were managed by surgery and chemotherapy. In a few MALT- omas which turned into high grade malignant tumors, the prognosis was poor. Acquired MALT may develop as a reaction to autoimmune disease and infection. Hyper-immune reaction and MALT hyperplasia under stimulation may result in myoepithelial sialadenitis and lead to MALT-oma of the salivary gland.

OBJECTIVETo explore the differences in sensitivity to rapamycin of five esophageal squamous cell carcinoma cell lines with different differentiation and the changes of sensitivity of the cells after siRNA-interfered expression of p70S6K.

METHODSEffects of rapamycin on proliferation of ESCC cell lines with different differentiation, EC9706, TE-1, Eca109, KYSE790 and KYSE450 cells, were investigated using cell counting kit 8 (CCK-8) assay, and according to the above results, the EC9706 cells non-sensitive to rapamycin were chosen to be transfected with p70S6K-siRNA. The changes in sensitivity of cells to rapamycin were measured in vitro and in vivo using CCK-8 kit, flow cytometry and tumor formation in nude mice.

RESULTSCCK-8 results showed that all the five cell line cells were sensitive to low concentration of rapamycin (<100 nmol/L), but TE-1 and EC9706 cells, which were with poor differentiation, showed resistance to high concentration of rapamycin. After EC9706 cells were treated with 50, 100, 200, 500 and 1 000 nmol/L rapamycin and p70S6K-siRNA, the proliferation rates of EC9706 cells were (48.67 ± 1.68)%, (15.45 ± 1.54)%, (14.00 ± 0.91)%, (10.97 ± 0.72)% and (2.70 ± 0.32)%, respectively, and were significantly lower than that of cells treated with 50, 100, 200, 500 and 1 000 nmol/L rapamycin and control siRNA [(74.53 ± 1.71)%, (68.27 ± 1.35)%, (71.74 ± 2.44)%, (76.23 ± 1.02)% and (80.21 ± 2.77)%] (P<0.05 for all). The results of flow cytometry showed that the ratios of cells in G1 phase of the p70S6K-siRNA, rapamycin and p70S6K-siRNA+ rapamycin groups were (53.82 ± 1.78)%, (57.87 ± 4.01)% and (73.73 ± 3.68)%, respectively, significantly higher than that in the control group (46.09 ± 2.31)% (P<0.05 for all). The results of tumor formation test in vivo showed that the inhibitory effect of rapamycin on tumor growth was stronger after the cells were transfected with p70S6K-siRNA, and the inhibition rate was 96.5%.

CONCLUSIONESCC cells with different differentiation have different sensitivity to rapamycin, and p70S6K-siRNA can improve the sensitivity of cells to rapamycin in vitro and in vivo.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; Carcinoma, Squamous Cell ; drug therapy ; metabolism ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; drug therapy ; metabolism ; pathology ; Humans ; Mice ; Mice, Nude ; RNA, Small Interfering ; Ribosomal Protein S6 Kinases, 70-kDa ; genetics ; metabolism ; Signal Transduction ; Sirolimus ; pharmacology ; Transfection

OBJECTIVETo investigate the relationship between specific immunocyte and pseudoanaphylactoid reactions (PR) induced by Shuanghuanglian injection (SHLI).

METHODICR mice, SCID mice and BALB/C athymic mouse were divided into different test groups, the mice were intravenously injected with solutions of different concentration of SHLI, positive control Compound 48/80 and normal sodium. All test substances were mixed with 0. 4% Evans blue. The reaction and vascular permeability of the ears were observed and measured 30 min after SHLI injected.

RESULTSHLI of 300, 600 mg x kg(-1) caused obvious vascular hyperpermeability in ICR mice, but the same dose of SHLI didn't cause vascular hyperpermeability in SCID mice and BALB/C athymic mouse.

CONCLUSIONSHLI in equivalent and 2 times the clinical dose can cause PR in ICR mice, but the same dose of SHLI can't cause PR in SCID mice and BALB/C athymic mouse, so specific immunocyte maybe take part in the SHLI-induced PR.

Anaphylaxis ; chemically induced ; immunology ; Animals ; Drugs, Chinese Herbal ; toxicity ; Immune System ; drug effects ; Injections ; methods ; Mice ; Mice, Inbred BALB C ; Mice, Inbred ICR ; Mice, SCID