Pharmacological activity and clinical progress of Triptolide and its derivatives LLDT-8, PG490-88Na, and Minnelide: a narrative review.

Triptolide, a compound isolated from a Chinese medicinal herb, has potent antitumor, immunosuppressive, and anti-inflammatory properties. Due to its interesting structural features and diverse pharmacological activities, it has attracted great interest by the Society of Organic Chemistry and Pharmaceutical Chemistry. However, its clinical potential is greatly hampered by limited aqueous solubility and oral bioavailability, and multi-organ toxicity. In recent years, various derivatives of Triptolide have made varying degrees of progress in the treatment of inflammatory diseases, autoimmune diseases, and cancer. The most researched and potentially clinically valuable of them were (5R)-5-hydroxytriptolide (LLDT-8), PG490-88Na (F6008), and Minnelide. In this review, we provide an overview of the advancements made in triptolide and several of its derivatives' biological activity, mechanisms of action, and clinical development. We also summarized some prospects for the future development of triptolide and its derivatives. It is hoped to contribute to a better understanding of the progress in this field, make constructive suggestions for further studies of Triptolide, and provide a theoretical reference for the rational development of new drugs.

[Interventional effect and mechanism of fermentation liquid of Dendrobium officinale leaves on alcoholic hepatitis mice].

Objective: To explore the intervention effect and mechanism of Dendrobium officinale leaf fermentation liquid on alcoholic hepatitis (AH) mice. Methods: Seventy inbred C57BL/6J male mice aged 6-8 weeks were selected and randomly divided into normal group (NG), model group (MG), liquid feed control group (CG), silybum group (SI), low-dose group (DL), medium-dose group (DM), and high-dose group (DH) of Dendrobium officinale fermentation liquid, with 10 mice in each group. NG group was given common feed, CG group was given control feed (LB alcoholic liquid control feed), SI group was given LB alcoholic liquid feed and silybum by gavage, DL, DM and DH groups were given LB alcoholic liquid feed and 25%, 50% and 100% concentration of Dendrobium officinale leaf fermentation liquid by gavage. An AH model was established by feeding LB alcoholic liquid feed for 8 weeks.At week 8, alanine Transaminase (ALT), triglyceride (TG), transferrin (TRF), interleukin (IL)-6, IL-10, and IL-1ß, tumor necrosis factor-α(TNF-α), interferon-γ(IFN-γ) were detected in eye blood of mice. Liver tissues were stained with HE, Oil Red O, Prussian blue and immunofluorescence ROS. The contents of glutathione(GSH) and malondialdehyde (MDA) in liver tissue homogenate were detected. To analyze the intervention effect and mechanism of Dendrobium officinale leaf fermentation solution on AH mice, the mRNA and protein relative expression levels of adenylate activated protein kinase (AMPK), AMPKß1, phosphorylated AMPKß1 (p-AMPKß1), tumor suppressor gene p53 (p53), solsolic vector family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GXP4) were detected by polymerase chain reaction (PCR) and Western blot. Results: Compared with MG group, the serum ALT and TG levels in the DL, DM, and DH groups were all reduced [ALT: (45.94±19.85), (45.73±22.62), and (41.68±7.13) vs (75.51±17.76) U/L, respectively; TG: (0.90±0.23), (0.69±0.22) and (0.41±0.20) vs (1.28±0.19) mmol/L, respectively, all P<0.05]; IL-6, IL-1ß, TNF-α, IFN-γ were decreased (all P<0.05). The serum TRF and IL-10 levels in the DM and DH groups were increased (all P<0.05). Compared with MG group, the liver tissue MDA of mice in DL, DM and DH groups was decreased [(0.41±0.05), (0.40±0.03), and (0.43±0.14) vs (0.64±0.06)µmol/g, respectively], GSH was increased (all P<0.05). Compared with MG, mRNA expression levels of AMPK (1.36±0.11, 1.61±0.17, 1.68±0.11 vs 0.80±0.12, respectively), SLC7A11 (0.91±0.12, 0.97±0.12, 0.99±0.13 vs 0.60±0.14, respectively) and GPX4 (0.51±0.11, 0.63±0.17, 0.83±0.15 vs 0.42±0.14, respectively) in the liver tissue of DL, DM and DH groups were all increased (all P<0.05). Compared with MG group, DL, DM and DH groups showed the relative expression levels of AMPKß1, p-AMPKß1, SLC7A11 and GPX4 were increased in the liver tissue of mice, while the relative expression levels of p53 protein were decreased (all P<0.05). Compared with MG group, DL, DM and DH groups reduced the degree of hepatic steatosis and inflammation in the lobules, while the iron and ROS staining in the liver tissue became lighter. Conclusion: Dendrobium officinale leaf fermentation liquid can alleviate the severity of AH in mice, and its mechanism may be related to the up-regulation of AMPK to inhibiting the p53/SLC7A11/GPX4 mediated Ferroptosis pathway.

Acupuncture for menopausal symptoms in Chinese women: a systematic review.

Menopause-related symptoms are common problems of middle-aged women that can seriously affect their quality of life. Menopausal hormone therapy (MHT) for climacteric symptoms is the first choice recommended by the International Menopause Society and likewise by other societies and institutions covering this field. However, non-hormonal therapies can be an alternative effective option, especially for women who are not suitable for MHT. Acupuncture is one of the most important methods. With deepening experience of the use of traditional Chinese acupuncture and moxibustion in the improvement of menopause symptoms, more clinical evidence has been obtained to support the effectiveness and safety of this treatment concept that is very often used in China. This review summarizes the evidence for effective treatment of climacteric complaints by acupuncture in recent years, shares the clinical experience of the authors of this review, all of whom head or work in units with daily large numbers of outpatients, and includes, in particular, results from studies performed in the Department of Acupuncture--Moxibustion of Tsinghua University Chuiyangliu Hospital, Beijing, China. In addition, there is a summary about the safety of acupuncture treatment in traditional Chinese medicine.

A single psychotomimetic dose of ketamine decreases thalamocortical spindles and delta oscillations in the sedated rat.

BACKGROUND: In patients with psychotic disorders, sleep spindles are reduced, supporting the hypothesis that the thalamus and glutamate receptors play a crucial etio-pathophysiological role, whose underlying mechanisms remain unknown. We hypothesized that a reduced function of NMDA receptors is involved in the spindle deficit observed in schizophrenia. METHODS: An electrophysiological multisite cell-to-network exploration was used to investigate, in pentobarbital-sedated rats, the effects of a single psychotomimetic dose of the NMDA glutamate receptor antagonist ketamine in the sensorimotor and associative/cognitive thalamocortical (TC) systems. RESULTS: Under the control condition, spontaneously-occurring spindles (intra-frequency: 10-16 waves/s) and delta-frequency (1-4 Hz) oscillations were recorded in the frontoparietal cortical EEG, in thalamic extracellular recordings, in dual juxtacellularly recorded GABAergic thalamic reticular nucleus (TRN) and glutamatergic TC neurons, and in intracellularly recorded TC neurons. The TRN cells rhythmically exhibited robust high-frequency bursts of action potentials (7 to 15 APs at 200-700 Hz). A single administration of low-dose ketamine fleetingly reduced TC spindles and delta oscillations, amplified ongoing gamma-(30-80 Hz) and higher-frequency oscillations, and switched the firing pattern of both TC and TRN neurons from a burst mode to a single AP mode. Furthermore, ketamine strengthened the gamma-frequency band TRN-TC connectivity. The antipsychotic clozapine consistently prevented the ketamine effects on spindles, delta- and gamma-/higher-frequency TC oscillations. CONCLUSION: The present findings support the hypothesis that NMDA receptor hypofunction is involved in the reduction in sleep spindles and delta oscillations. The ketamine-induced swift conversion of ongoing TC-TRN activities may have involved at least both the ascending reticular activating system and the corticothalamic pathway.

[Distribution of allergens and associated risk factors of patients with allergic rhinitis in Qingdao area].

Objective:To analyze the distribution of allergens and the risk factors in patients with allergic rhinitis in Qingdao area. Method:Two hundred and sixteen patients diagnosed with allergic rhinitis and 49 healthy volunteers were admitted to the outpatient clinic of the Otolaryngology Department of the three branches of Qingdao University Affiliated Hospital in Qingdao from August 2018 to March 2019. A total of 271 patients（5-84 years old） were included in the epidemiological investigation. The study conducted a skin stinging test of inhalation allergens. In addition, the questionnaire of allergic disease and self-examination of dust mite allergy were carried out for each group of patients, and the results were analyzed and counted. Result:①In 216 patients with allergic rhinitis, the top six inhalation allergens were dust mites（55.0%）, house dust mites（46.1%）, tropical mites（21.8%）, dog hair（16.6%）, Artemisia halodendron（13.3%）, and German cockroaches（10.3%）. ②Inhalation allergens, the degree of positive of house dust mites and dust mites were mainly（+++）, the degree of positive of Artemisia halodendron were mainly（+++） and（++++）, and the degree of positive of other allergens were mainly（++）. ③There were statistically significant differences in the positive rates of inhalation allergens in different age groups, and the positive rates of inhalation allergens gradually decreased with age after adulthood. ④The single factor and multi-factor logical regression analysis all showed that the history of smoking, family history, age less than 18 years, and 18-40 years old were closely related to the occurrence of allergic rhinitis（P<0.05）. ⑤Using Spearson-related analysis and loess regression analysis, it was found that with the increase of dust mite self-examination score, the positive rate of dust mite increased significantly. Conclusion:The most important inhalation allergens of patients with allergic rhinitis in Qingdao area are dust mites, dog hair and artemisinin. The positive rate of inhalation allergens is influenced by age and there are certain rules in them. Smoking history, family history, age and other factors are risk factors for allergic rhinitis. The dust mite self-examination score can predict the occurrence of dust mite allergy.

Deep insemination with sex-sorted Cashmere goat sperm processed in the presence of antioxidants.

Flow cytometrically sex-sorted sperm have been widely used for improving reproductive management in the dairy industry. However, the industrial application of this technology in other domestic species is largely limited by the lower fertility after insemination. The aim of this study was to investigate effects of antioxidant supplementation during the sex-sorting and freezing process on the quality and functions of sorted sperm from Liaoning Cashmere goats. We tested the effects of antioxidant supplementation during sex-sorting and freezing process, including ascorbic acid-2-glucoside AA-2G, glutathione, melatonin and vitamin C (VC), on the quality and functions of sex-sorted fresh and frozen-thawed sperm. Based on these experiments, we performed deep insemination with sex-sorted sperm using our improved strategy, in comparison to unsorted sperm. In Experiment 1, compared with control group and other antioxidants, AA-2G supplementation significantly alleviated the degradation of motility and viability of fresh sperm after sorting and showed the highest percentage of sperm with normal morphology. In addition, AA-2G supplementation showed an evident protection against the sorting process-induced membrane and acrosome damage. In Experiment 2, AA-2G supplementation was most effective in protecting motility, while melatonin supplementation appears to facilitate the degradation of quality of frozen-thawed sex-sorted sperm. In Experiment 3, we performed deep insemination with sperm that were sorted and frozen in the presence of AA-2G and obtained a satisfying pregnancy rate comparable to that from unsorted sperm. The results showed that AA-2G supplementation efficiently protects quality and function of both fresh and frozen-thawed sex-sorted sperm of Cashmere goats, thus obtaining a satisfying pregnancy outcome.

RN1, a novel galectin-3 inhibitor, inhibits pancreatic cancer cell growth in vitro and in vivo via blocking galectin-3 associated signaling pathways.

Galectin-3 (Gal-3) has been implicated in pancreatic ductal adenocarcinoma (PDAC), and its candidacy as a therapeutic target has been evaluated. Gal-3 is widely upregulated in tumors, and its expression is associated with the development and malignancy of PDAC. In the present study, we demonstrate that a polysaccharide, RN1, purified from the flower of Panax notoginseng binds to Gal-3 and suppresses its expression. In addition, RN1 markedly inhibits PDAC cells growth in vitro, in vivo and in patient-derived xenografts. Mechanistically, RN1 binds to epidermal growth factor receptor (EGFR) and Gal-3, thereby disrupting the interaction between Gal-3 and EGFR and downregulating extracellular-related kinase (ERK) phosphorylation and the transcription factor of Gal-3, Runx1 expression. Inhibiting the expression of Runx1 by RN1, suppresses Gal-3 expression and inactivates Gal-3-associated signaling pathways, including the EGFR/ERK/Runx1, BMP/smad/Id-3 and integrin/FAK/JNK signaling pathways. In addition, RN1 can also bind to bone morphogenetic protein receptors (BMPR1A and BMPR2) and block the interaction between Gal-3 and the BMPRs. Thus, our results suggest that a novel Gal-3 inhibitor RN1 may be a potential candidate for human PDAC treatment via multiple targets and multiple signaling pathways.

[Two cases with generalized intracranial calcification due to hereditary folate malabsorption and literature review].

Objective: This study aimed to investigate the clinical, biochemical and genetic features of two Chinese children with hereditary folate malabsorption. Method: Clinical features, laboratory examinations, treatment and SLC46A1 gene of two cases were studied. Reports on hereditary folate malabsorption utill September of 2016 were searched and the clinical and genetic characteristics of reported cases were summarized. Result: The two patients presented with megaloblastic anemia from their infant period and seizures, psychomotor retardation and regression. In case1, mean corpuscular volume (MCV) was 100 fl. Serum folate was 9.96 nmol/L. Folate and 5-methylenetetrahydrofolate in cerebrospinal fluid were 0 and 0.01 separately. In case 2, MCV was 93.9 fl. Serum folate was 4.49 nmol/L. The concentration of folate and 5-methylenetetrahydrofolate in cerebrospinal fluid were both zero. On their brain CT, progressive bilateral symmetrical calcification was observed. On their SLC46A1 gene, four mutations were identified. Case 1 had one novel mutation, c. 1238T>C (L413P) and c. 194-195insG (p.Cys66LeufsX99). From Case 2, two reported mutations, c. 1A>T (M1L) and c. 194-195insG (p.Cys66LeufsX99) were identified. The administration of folinic acid (60 to 120 mg per day) was initiated after diagnosis. Clinical improvement and normalized hematologic markers were observed after treatment. Totally 37 cases were reported in reviewed English literature, including 30 cases with mutations on SLC46A1 gene (only one Chinese patient). All the cases had the onset in infancy. The ratio of boys to girls was 1 to 1.5. Main manifestations were characterized by megaloblastic anemia (77%), failure to thrive (50%), diarrhea (27%), psychomotor retardation (63.6%), epilepsy (27%), and infection of respiratory system (45.5%). The concentration of folate in both serum and cerebrospinal fluid was decreased (72.7% and 63.6% respectively). Hypoimmunoglobulinemia accounted for 27.3%. Most of mutations in HFM were distributed between p. 65 and p. 68 (c.194-c.204), mainly due to insertion- or deletion-related frame shifts or generation of stop codons. Oral and parenteral folinic acid treatment was effective. Conclusion: Hereditary folate malabsorption often presented with megaloblastic anemia, abnormalities of digestive and nervous system, and hypoimmunoglobulinemia with recurrent infections. Low level of serum and CSF folate and screening SLC46A1 gene are keys to the etiologic study of the patients. Early supplement with folinic acid is beneficial to the prognosis.

Production of succinic acid from sugarcane molasses supplemented with a mixture of corn steep liquor powder and peanut meal as nitrogen sources by Actinobacillus succinogenes.

The potential of using corn steep liquor powder (CSLP), peanut meal (PM), soybean meal (SM), cotton meal (CM) and urea as the substitute of yeast extract (YE) as the nitrogen source was investigated for producing succinic acid (SA). Actinobacillus succinogenes GXAS137 was used as the fermenting bacterium and sugarcane molasses was used as the main substrate. None of these materials were able to produce SA as high as YE did. The CSLP could still be considered as a feasible and inexpensive alternate for YE as the yield of SA produced using CSLP was second only to the yield of SA obtained by YE. The use of CSLP-PM mixed formulation (CSLP to PM ratio = 2·6) as nitrogen source produced SA up to 59·2 g l(-1) with a productivity of 1·2 g l(-1) h(-1). A batch fermentation using a stirred bioreactor produced up to 60·7 g l(-1) of SA at the same formulation. Fed-batch fermentation that minimized the substrate inhibition produced 64·7 g l(-1) SA. These results suggest that sugarcane molasses supplemented with a mixture of CSLP and PM as the nitrogen source could be used to produce SA more economically using A. succinogenes. Significance and impact of the study: Succinic acid (SA) is commonly used as a platform chemical to produce a number of high value derivatives. Yeast extract (YE) is used as a nitrogen source to produce SA. The high cost of YE is currently the limiting factor for industrial production of SA. This study reports the use of a mixture of corn steep liquor powder (CSLP) and peanut meal (PM) as an inexpensive nitrogen source to substitute YE. The results showed that this CSLP-PM mixed formulation can be used as an effective and economic nitrogen source for the production of SA.

Genetic diversity of endangered Polyporus umbellatus from China assessed using a sequence-related amplified polymorphism technique.

Polyporus umbellatus (Pers.) Fries is an endangered medicinal fungus in China with in vivo anticancer activity, but its genetic information is lacking. Eight natural P. umbellatus strains collected from 7 provinces in China were subjected to sequence-related amplified polymorphism markers to estimate the level and pattern of genetic diversity. Forty-nine primer combinations generated 1219 highly reproducible and discernible loci, among which 1023 were polymorphic. The percentage of polymorphism varied from 35.71 to 96.30 with an average of 83.92. Genetic identity among all strains ranged from 0.15 to 0.78 with an average of 0.46. The unweighted pair group method with arithmetic mean dendrogram clustered 8 strains into 3 clusters, and the clustering pattern showed 3 groups. Principal coordinate analysis further indicated that the genetic diversity of P. umbellatus strains was unevenly distributed and instead displayed a clustered distribution pattern. A relatively high level of genetic diversity was maintained in 8 natural P. umbellatus strains, but its abundance might be subjected to environmental heterogeneity, and the population structure of co-evolved Armillaria species may be selected by nature under the specific microenvironment.

Modulation of growth and angiogenic potential of oral squamous carcinoma cells in vitro using salvianolic acid B.

BACKGROUND: Our previous studies showed that Salvianolic acid B (Sal B) inhibited 7,12-dimethylbenz[a]anthracene (DMBA)-induced oral carcinogenesis in hamsters and such anti-cancer effects might be related to the inhibition of angiogenesis. This study was aimed to further investigate the anti-proliferative effect of Sal B on the most common type of oral cancer, oral squamous cell carcinoma (OSCC) and the possible mechanisms of action with respect to angiogenesis inhibition. METHODS: Two well-characterized oral squamous cell carcinoma cell lines, CAL27 and SCC4, and premalignant leukoplakia cells were treated with different concentrations of Sal B. Cytotoxicity was assessed by MTT assay. cDNA microarray was utilized to evaluate the expression of 96 genes known to be involved in modulating the biological processes of angiogenesis. Real-time reverse transcription-polymerase chain reaction analysis was conducted to confirm the cDNA microarray data. RESULTS: Sal B induced growth inhibition in OSCC cell lines but had limited effects on premalignant cells. A total of 17 genes showed a greater than 3-fold change when comparing Sal B treated OSCC cells to the control. Among these genes, HIF-1α, TNFα and MMP9 are specifically inhibited, expression of THBS2 was up-regulated. CONCLUSIONS: Sal B has inhibitory effect on OSCC cell growth. The antitumor effect can be attributed to anti-angiogenic potential induced by a decreased expression of some key regulator genes of angiogenesis. Sal B may be a promising modality for treating oral squamous cell carcinoma.

Whole body radioprotective activity of an acetone-water extract from the seedpod of Nelumbo nucifera Gaertn. seedpod.

Procyanidins extracted with acetone-water from lotus (Nelumbo nucifera Gaertn.) seedpod (LSPCs) were evaluated for in vivo radioprotective activity against whole body gamma irradiation in Swiss albino mice. Pretreated with LSPCs 200 mg/kg by intragastric (i.g.) for 15 days was found to be the most effective dose in preventing radiation sickness, reducing radiation-induced mortality, increasing mean survival time and elevating radiation median lethal dose (LD(50)) from 8.9 to 10.5 Gy, indicating a dose modifying factor (DMF) of 1.18. Further, administered LSPCs at a dose of 200 mg/kg could effectively maintain spleen index close to normal, stimulate endogenous spleen colony forming units, promote the levels of red blood cells (RBC), white blood cells (WBC), platelets and hemoglobin in peripheral blood, and prevent spleen and skin damage in irradiated mice, reduce the level of radiation-induced micronucleated polychromatic erythrocytes in bone marrow, maintain the polychromatic erythrocytes (PCE) and normochromatic erythrocytes (NCE) ratio (P/N ratio) and significantly decrease bone marrow chromosomal damage. Alternatively, pretreated with LSPCs (200 mg/kg) significantly decreased the lipid peroxidation (LPO) level, and elevated the activities of endogenous antioxidant enzymes in liver after irradiation. Thus LSPCs possess a strong whole body radioprotective activity, and it may be used as a radioprotector.

Dynamic skeletal muscle stimulation and its potential in bone adaptation.

To identify mechanotransductive signals for combating musculoskeletal deterioration, it is essential to determine the components and mechanisms critical to the anabolic processes of musculoskeletal tissues. It is hypothesized that the interaction between bone and muscle may depend on fluid exchange in these tissues by mechanical loading. It has been shown that intramedullary pressure (ImP) and low-level bone strain induced by muscle stimulation (MS) has the potential to mitigate bone loss induced by disuse osteopenia. Optimized MS signals, i.e., low-intensity and high frequency, may be critical in maintaining bone mass and mitigating muscle atrophy. The objectives for this review are to discuss the potential for MS to induce ImP and strains on bone, to regulate bone adaptation, and to identify optimized stimulation frequency in the loading regimen. The potential for MS to regulate blood and fluid flow will also be discussed. The results suggest that oscillatory MS regulates fluid dynamics with minimal mechanical strain in bone. The response was shown to be dependent on loading frequency, serving as a critical mediator in mitigating bone loss. A specific regimen of dynamic MS may be optimized in vivo to attenuate disuse osteopenia and serve as a biomechanical intervention in the clinical setting.

The effects of Rhodiola rosea extract on 5-HT level, cell proliferation and quantity of neurons at cerebral hippocampus of depressive rats.

The purpose of this study was to investigate the effects of Rhodiola rosea extract and depression on the serotonin (5-HT) level, cell proliferation and quantity of neurons at cerebral hippocampus of depressive rats induced by Chronic Mild Stress (CMS). Seventy male Sprague-Dawley rats were divided into seven groups (10 per group): normal control group, untreated depressive rat model group, negative control group, positive control group, low dosage Rhodiola rosea extract (1.5g/kg) group, medium dosage Rhodiola rosea extract (3g/kg) group and high dosage Rhodiola rosea extract (6g/kg) group. After the depressive rats induced by CMS had received Rhodiola rosea extract for 3 weeks, the 5-HT levels at cerebral hippocampus were detected by high performance liquid chromatography. Bromodeoxyuridine (BrdU) was injected in vivo to label the proliferating cells at hippocampus, and morphometry was used to count the hippocampal neurons. The results showed that the 5-HT level of the three experimental groups had recovered to normal status. The immunohistochemistry of hippocampus BrdU positive cells had returned to the normal level in the group of depressive rats with low dosage Rhodiola rosea extract. In conclusion the results demonstrated that Rhodiola rosea extract could improve 5-HT level in hippocampus in depressive rats, and low dosage Rhodiola rosea could induce neural stem cell proliferation at hippocampus to return to normal level, repairing the injured neurons at hippocampus.

Histological evaluation of the safety of toluidine blue-mediated photosensitization to periodontal tissues in mice.

Our previous in vitro study has shown that toluidine blue (TB)-mediated lethal photosensitization of periodontal pathogens (PPs) from periodontal patients is possible. The purpose of this study was to investigate whether TB-mediated photosensitization exerted damaging effects on periodontal tissues in mice. Twenty-four mice were randomly divided into four groups; the experimental photodynamic therapy (PDT) group was treated with 1 mg/ml TB and light irradiation (60 J/cm(2), 635 nm, 337 s). Those in control groups were subjected to 140 J/cm(2) laser irradiation alone or to 2.5 mg/ml TB alone or received neither TB nor light exposure. All the mice were killed 72 h after they had been subjected to PDT, and periodontal tissue samples were taken for histological examination. During the 72 h observation period, no mice showed any distress. No necrotic or inflammatory changes were found in the gingiva, dentin, dental pulp or alveolar bone of the mice in any of the groups in this study. The results suggest that TB-mediated PDT is a safe antimicrobial approach for the treatment of periodontopathy without damaging effects to adjacent normal tissues.

Localization of arabinogalactan proteins in anther, pollen, and pollen tube of Nicotiana tabacum L.

Western blot analysis indicated the presence of two epitopes recognized by the anti-arabinogalactan protein antibodies JIM13 and LM2 and the absence of the JIM4 epitope in mature tobacco anthers. Immunoenzyme localization of arabinogalactan proteins (AGPs) with JIM13 showed that AGPs accumulate mainly at the early stages of anther development. AGP content and distribution were also investigated at the ultrastructural level in pollen tubes grown in vivo and in vitro. Abundant AGPs were present in the transmitting tissue of styles, and the AGP content of the extracellular matrix changed during pollen tube growth. In pollen tubes, immunogold particles were mainly distributed in the cell wall and cytoplasm, especially around the peripheral region of the generative-cell wall. beta-D-Glucosyl Yariv reagent, which specifically binds to AGPs, caused slow growth of pollen tubes and reduced immunogold labeling of AGPs with JIM13 in vitro. These data suggest that AGPs participate in male gametogenesis and pollen tube growth and may be important surface molecules in generative and sperm cells.

Cytotoxicity induced by grape seed proanthocyanidins: role of nitric oxide.

Grape seed proanthocyanidin extract (GPSE) at high doses has been shown to exhibit cytotoxicity that is associated with increased apoptotic cell death. Nitric oxide (NO), being a regulator of apoptosis, can be increased in production by the administration of GSPE. In a chick cardiomyocyte study, we demonstrated that high-dose (500 microg/ml) GSPE produces a significantly high level of NO that contributes to increased apoptotic cell death detected by propidium iodide and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. It is also associated with the depletion of intracellular glutathione (GSH), probably due to increased consumption by NO with the formation of S-nitrosoglutathione. Co-treatment with L-NAME, a NO synthase inhibitor, results in reduction of NO and apoptotic cell death. The decline in reduced GSH/oxidized GSH (GSSG) ratio is also reversed. N-Acetylcysteine, a thiol compound that reacts directly with NO, can reduce the increased NO generation and reverse the decreased GSH/GSSG ratio, thereby attenuating the cytotoxicity induced by high-dose GSPE. Taken together, these results suggest that endogenous NO synthase (NOS) activation and excessive NO production play a key role in the pathogenesis of high-dose GSPE-induced cytotoxicity.

[Locating the impairment of human cognitive function during hypoxia].

Objective. To find the location where the human brain cognitive function impairs under hypoxia. Method. 14 healthy males, aged 18-20 years, performed auditory Oddball test of two different intensities (55 dB, 80 dB) during exposure to 5000 m by breathing low oxygen mixture. EEG and Reaction Time (RT) were recorded. P3, the component of ERP, was extracted from EEG. P3 latency and RT were used as indices of the experiment. Additive Factors Method was used to analyse the result of the experiment. Result. Interaction between hypoxia and stimulus intensity was found for P3 latency and RT. Conclusion. Acute hypoxia influences the preprocessing stage of information processing.

[Effect of conventionally prepared rabbit serum on human umbilical vein endothelial cell-304].

OBJECTIVE: To study the effect of conventionally prepared rabbit serum, usually used in the field of serum pharmacology of TCM, in various concentration on cultured system of human umbilical vein endothelial cell (VEC)-304 cell line. METHODS: Prepared conventionally from healthy rabbit, the effect of uncontaminated and nonhemolytic serum of six concentrations (5%, 10%, 20%, 40%, 60% and 80%) on VEC-304 cells was observed using parameters such as cell morphologic examination, trypan blue exclusion test, methyl thiazolyl tetrazolium (MTT) test, lactate dehydrogenase (LDH) and angiotensin-converting release test. RESULTS: Serum of 5% and 10% concentration showed no effect in all the above-mentioned tests. Serum of 20% concentration also showed no effect in the tests except for LDH releasing (P < 0.01). But when the concentration of serum reached up to 40% or above, distinct cytotoxicity and serious injury appeared in a dose-dependent manner. CONCLUSION: High concentration of conventionally prepared rabbit serum could injure the incubated cell. Therefore, in order to lower the background effect of serum as far as possible in TCM serum pharmacological study, the cytotoxicity of rabbit serum should be tested first of all.

Determination of edible bird's nest and its products by gas chromatography.

A specific gas chromatographic (GC) detection method for edible bird's nest (EBN) based on identifying the composition of the oligosaccharide chain combined with glycoprotein in EBN is developed. Five monoses (D-mannitose, D-galactose, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, and N-acetyl neuraminate) that constitute the oligosaccharide chain are detected using GC and GC-mass spectrometry techniques; their characteristic GC spectrum can reliably be regarded as EBN's fingerprint. The peak-area ratios in GC spectrum of those five monoses are found to be fixed; therefore, the GC technique developed in this work can conveniently be used to determine various raw EBNs and their products both qualitatively and quantitatively, distinguishing between fake and genuine EBN rapidly.