[Alteration and significance of serum lipid levels and nutritional status during BCMA-CAR-T-cell therapy in patients with refractory or relapsed multiple myeloma: a retrospective study based on LEGEND-2].

Objective: To explore the dynamic changes in serum lipid levels and nutritional status during BCMA-CAR-T-cell therapy in patients with refractory or relapsed multiple myeloma (R/R MM) based on LEGEND-2. Methods: The data of patients with R/R MM who underwent BCMA-CAR-T therapy at our hospital between March 30, 2016, and February 6, 2018, were retrospectively collected. Serum lipid levels, controlled nutritional status (CONUT) score, and other clinical indicators at different time points before and after CAR-T-cell infusion were compared and analyzed. The best cut-off value was determined by using the receiver operator characteristic (ROC) curve. The patients were divided into high-CONUT score (>6.5 points, malnutrition group) and low-CONUT score groups (≤6.5 points, good nutrition group), comparing the progression-free survival (PFS) and total survival (OS) of the two groups using Kaplan-Meier survival analysis. Results: Before the infusion of CAR-T-cells, excluding triglycerides (TG), patients' serum lipid levels were lower than normal on average. At 8-14 d after CAR-T-cell infusion, serum albumin (ALB), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and apolipoprotein A1 (Apo A1) levels dropped to the minimum, whereas CONUT scores reached the maximum. In addition to TG, apolipoprotein B (Apo B) levels increased compared with baseline. After CAR-T-cell therapy, the patients' serum lipid levels significantly increased with well-improved nutritional status. Spearman's related analysis showed that TC, HDL, and ApoA1 levels after CAR-T-cell injection were significantly negatively correlated with the grade of cytokine-release syndrome (CRS) (r=-0.548, P=0.003; r=-0.444, P=0.020; r=-0.589, P=0.001). Furthermore, survival analysis indicated that the CONUT score was unrelated to PFS, and the median OS of patients with R/R MM in the high-CONUT score group was shorter than that in the low-CONUT score group (P=0.046) . Conclusions: During CAR-T-cell therapy, hypolipidemia and poor nutritional status were aggravated, which is possibly related to CRS. The patients' serum lipid levels and nutritional status were significantly improved after CAR-T-cell treatment. The CONUT score affected the median OS in patients treated with CAR-T-cells. Therefore, specific screening and intervention for nutritional status in patients receiving CAR-T-cell therapy are required.

Effect of antioxidant procyanidin b2 (pcb2) on ovine oocyte developmental potential in response to in vitro maturation (ivm) and vitrification stress.

BACKGROUND: It was demonstrated that external stress, such as in vitro maturation (IVM) and vitrification process can induce significantly reduced development capacity in oocytes. Previous studies indicated that antioxidants play a pivotal part in the acquisition of adaptation in changed conditions. At present, the role of the natural potent antioxidant PCB2 in response to IVM and vitrification during ovine oocyte manipulation has not been explored. OBJECTIVE: To investigate whether PCB2 treatment could improve the developmental potential of ovine oocytes under IVM and vitrification stimuli. MATERIALS AND METHODS: The experiment was divided into two parts. Firstly, the effect of PCB2 on the development of oocytes during IVM was evaluated. Un-supplemented and 5 ug per mL PCB2-supplemented in the IVM solution were considered as control and experimental groups (C + 5 ug per mL PCB2). The polar body extrusion (PBE) rate, mitochondrial membrane potential (MMP), ATP, reactive oxygen species (ROS) levels and early apoptosis of oocytes were measured after IVM. Secondly, we further determine whether PCB2 could improve oocyte quality under vitrification stress. The survival rate, PBE rate and early apoptosis of oocytes were compared between fresh group, vitrified group and 5 ug per mL PCB2-supplemented in the IVM solution after vitrification (V + 5 ug per mL PCB2). RESULTS: Compared to the control group, adding PCB2 significantly increased PBE rate (79.4% vs. 62.8%, P < 0.01) and MMP level (1.9 +/- 0.08 vs. 1.3 +/- 0.04, P < 0.01), and decreased ROS level (47.1 +/- 6.3 vs. 145.3 +/- 8.9, P < 0.01). However, there was no significant difference in ATP content and early apoptosis. Compared to the fresh group, vitrification significantly reduced oocytes viability (43.0% vs. 90.8%, P < 0.01) as well as PBE rate (24.2% vs. 60.6%, P < 0.05). However, 5 ug per mL PCB2-supplemention during maturation had no effect on survival, PBE or early apoptosis in vitrified oocytes. CONCLUSION: PCB2 could effectively antagonise the oxidative stress during IVM and promote oocyte development. DOI: 10.54680/fr23210110412.

Neobavaisoflavone improves medial collateral ligament-induced osteoarthritis through repressing the nuclear factor -κB/hypoxia-inducible factor-2α axis.

Osteoarthritis (OA) is a chronic inflammatory joint disease. There have been some studies on the treatment of OA with traditional Chinese medicine (TCM). Neobavaisoflavone (NBIF) is an isoflavone isolated from TCM Psoralea corylifolia L (also called 'Buguzhi') and shows anti-inflammatory effects. This study aims to explore the potential role of NBIF in treating OA. The rat chondrocytes were dealt with interleukin-1beta (IL-1ß) for inducing an in-vitro OA model and a rat OA model was established by medial collateral ligament resection. Followed by NBIF treatment, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry were performed to evaluate chondrocyte proliferation and apoptosis. The expression of inflammatory factors and oxidative stress factors in chondrocyte medium and rat serum was tested by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining and Safranin O-Fast Green staining were carried out to examine the histopathological changes in knee joints. Caspase-3, nuclear factor-kappaB (NF-κB), and hypoxia-inducible factor-2alpha (HIF-2α) expressions were monitored by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blot, and/or immunohistochemistry. As indicated by the results, NBIF mitigated cartilage matrix degradation and chondrocyte apoptosis in the OA rat model. NBIF hampered IL-1ß-mediated cell viability inhibition, apoptosis, inflammatory reactions, and oxidative stress of chondrocytes. Moreover, NBIF suppressed NF-κB phosphorylation and HIF-2α expression. HIF-2α overexpression induced inflammation, oxidative stress, and apoptosis in chondrocytes, while NBIF reversed HIF-2α overexpression-caused chondrocyte damage. Overall, NBIF had antiapoptotic, anti-inflammatory, and antioxidative stress effects in OA models by impeding NF-κB/HIF-2α axis, suggesting that NBIF has potential therapeutic effects in OA.

Effects of Lonicera confusa and Astragali Radix extracts supplementation on egg production performance, egg quality, sensory evaluation, and antioxidative parameters of laying hens during the late laying period.

Lonicera confusa (LC) and Astragali Radix (AR) extracts have been shown to have antioxidant and anti-inflammatory activities in human. To determine whether LC or/and AR extracts had similar functions to improve the egg production and egg quality, 1,440 (52-wk-old) Lohmann pink-shell hens were randomly distributed into 4 treatments with nine replicates of 40 hens. The hens were fed a basal diet (CON) or the basal diet supplemented with 0.1% LC extracts, 0.1% AR extracts, or 0.1% LC extracts plus 0.1% AR extracts (LC-AR) for 12 wk. The eggs were collected on week 6 and 12 for analysis, and the plasma and ovaries were collected at end of trial. Dietary treatment did not influence (P < 0.05) egg production, egg weight, and feed conversion ratio. However, LC-AR addition increased (P < 0.02) yolk color and sensory quality of hard-boiled eggs compared to other groups. The LC-AR supplementation increased (P = 0.02), and LC addition tended to increase (P = 0.08) Haugh unit of eggs on week 12 compared to CON. The LC-AR supplementation decreased (P < 0.001), and LC or AR individual addition tended to decrease (P < 0.10) plasma malondialdehyde concentration compared to CON. Conversely, LC supplementation increased (P = 0.02) total superoxide dismutase activity, and LC or/and AR supplementation increased the activities of manganese-containing superoxide dismutase (MnSOD) (P < 0.08) and glutathione peroxidase (GSH-Px) (P < 0.01) in plasma, and the mRNA abundance of MnSOD, GSH-Px1, and catalase in ovaries (P < 0.05) compared to CON. The LC or/and AR supplementation decreased the concentrations of interleukin (IL)-6 and tumor necrosis factor alpha (TNF-α) in plasma (P < 0.05), and the mRNA abundance of IL-6 and TNF-α in ovaries (P < 0.04) compared to CON. These results suggested dietary inclusion of LC or/and AR improved the albumen quality, and a blend of LC and AR improved yolk color, which were associated with the enhancement of antioxidant capacity and the suppression of systemic inflammation in hens.

Effects of N-carbamylglutamate and L-arginine on gonadotrophin-releasing hormone (GnRH) gene expression and secretion in GT1-7 cells.

Recent studies have shown that N-carbamylglutamate (NCG) and arginine (ARG) supplementation improves reproductive performance in livestock. The objectives of the present study were to evaluate the effects of NCG and ARG on GT1-7 cell gonadotrophin-releasing hormone (GnRH) secretion, gene expression and cell proliferation. GT1-7 cells were treated in vitro with different concentrations of NCG (0-1.0mM) or ARG (0-4.0mM) in serum-free medium for 12 or 24h. For GnRH secretion and cell proliferation, GT1-7 cells were more sensitive to NCG than ARG. NCG treatment after 12h increased cell numbers and inhibited GnRH secretion in a dose-dependent manner (P<0.05), although there was no significant effect of NCG on these parameters after 24h culture. ARG treatment decreased GnRH secretion after 24h (P<0.05), whereas it had no effect after 12h. GT1-7 cells express GnRH, Kiss-1 metastasis-suppressor (Kiss1), G-protein coupled receptor 54 (GPR54), neuronal nitric oxide synthase (nNOS) and estrogen receptor α (ERα) genes. High concentrations of NCG (1.0mM) and ARG (4.0mM) inhibited (P<0.05) GnRH and nNOS mRNA abundance in GT1-7 cells. ARG treatment decreased Kiss1 and increased ERα mRNA abundance. Thus, high concentrations of NCG (1.0mM) and ARG (4.0mM) may act both directly and indirectly to regulate GnRH neuron function by downregulating genes related to GnRH synthesis and secretion to slow GnRH production while stimulating GT1-7 cell proliferation.

Effect of Shenqin biochemical extract on hypoxia-inducible factor-1α expression in ultraviolet B-irradiated HaCaT cells.

Hypoxia-inducible factor-1α (HIF-1α) is considered the main transcriptional regulator of the hypoxia-specific cellular and developmental response. This study was performed to investigate the effect of Shenqin biochemical extract (SQBE) on HIF-1α expression in ultraviolet B (UVB)-irradiated HaCaT cells and the possible action mechanisms of SQBE against UVB-induced skin cancer. HaCaT cells in logarithmic growth phase were seeded in Dulbecco's modified Eagle's medium with 10% fetal bovine serum, and conventionally cultured at 37°C with 5% CO2. Cells were divided into control group (administered the same amounts of dimethyl sulfoxide), SQBE1 group (12.5 µg/mL SQBE), SQBE2 group (25.0 µg/mL SQBE), and SQBE3 group (50.0 µg/mL SQBE). Four hours post administration, the control and treatment groups were irradiated with UVB (0, 20, 40, and 60 mJ/cm2). After 24 h, cell survival rate was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Expression levels of HIF-1α mRNA and protein were detected by polymerase chain reaction and western blotting, respectively. SQBE-treated, UVB-irradiated cells had improved survival rates. This increase was most significant in SQBE3 group (P < 0.01), which also had effectively reduced expression of intracellular HIF-1α mRNA and protein. Hence, SQBE had a protective effect on UVB-irradiated HaCaT cells and inhibited the UVB irradiation-induced expression of HIF-1α. This indicates that SQBE could prevent the occurrence of UVB radiation-induced skin cancer.

Effect of alpha-lipoic acid on relieving ammonia stress and hepatic proteomic analyses of broilers.

Ammonia in poultry houses not only affects worker health but also induces a variety of poultry diseases. Alpha-lipoic acid (LA) is an effective antioxidant that protects cells against oxidative injury during various toxic and pathological processes. This study was designed to evaluate the mitigating effects of LA supplementation on ammonia stress and hepatic proteome changes in broilers. Male broilers (22 d old) were allocated to 3 groups: (1) a control group without ammonia stress (CTRL); (2) exposure to 70 ppm ammonia (AM); and (3) exposure to 70 ppm ammonia and dietary administration of 300 mg/kg LA (AM+LA). Ammonia exposure significantly decreased broiler growth performance and plasma glutathione peroxidase activity (P < 0.05), and increased plasma malondialdehyde content and glutamic-pyruvic transaminase activity (P < 0.05). These negative effects were eliminated by LA supplementation. Comparative proteomic analyses revealed 291 differentially expressed proteins in the AM group compared to the CTRL and AM+LA groups. A total of 30 proteins were differentially expressed between the AM/CTRL and (AM+LA)/AM groups. The addition of LA restored 24 of these proteins to control levels; these proteins were mainly related to transcription regulation, detoxification, protein translation and degradation, and immune and stress responses. The differentially expressed proteins included the high mobility group box (HMGB) and glutathione S-transferase (GST), which is closely related to immune response and oxidative stress, and collagens, which are implicated in liver injury. The addition of LA to broiler diet may reduce ammonia toxicity by maintaining the antioxidant system, xenobiotic metabolism, and metabolic pathways.

Polycatechol nanosheet: a superior nanocarrier for highly effective chemo-photothermal synergistic therapy in vivo.

The integration of phototherapy and chemotherapy in a single system holds great promise to improve the therapeutic efficacy of tumor treatment, but it remains a key challenge. In this study, we describe our recent finding that polycatechol nanosheet (PCCNS) can be facilely prepared on a large scale via chemical polymerization at 4 °C, as an effective nanocarrier for loading high-density CuS nanocrystals as a photothermal agent. The resulting CuS/PCCNS nanocomposites exhibit good biocompatibility, strong stability, and a high photothermal conversion efficiency of ∼45.7%. The subsequent loading of anticancer drug doxorubicin (Dox) creates a superior theranostic agent with pH- and heat-responsive drug release, leading to almost complete destruction of mouse cervical tumor under NIR laser irradiation. This development offers an attractive theranostic agent for in vivo chemo-photothermal synergistic therapy toward biomedical applications.

Effects of chlortetracycline and copper supplementation on antimicrobial resistance of fecal Escherichia coli from weaned pigs.

Feed-grade chlortetracycline (CTC) and copper are both widely utilized in U.S. pig production. Cluster randomized experiment was conducted to evaluate the effects of CTC and copper supplementation in weaned pigs on antimicrobial resistance (AMR) among fecal Escherichia coli. Four treatment groups: control, copper, CTC, or copper plus CTC were randomly allocated to 32 pens with five pigs per pen. Fecal samples were collected weekly from three pigs per pen for six weeks. Two E. coli isolates per fecal sample were tested for phenotypic and genotypic resistance against antibiotics and copper. Data were analyzed with multilevel mixed effects logistic regression, multivariate probit analysis and discrete time survival analysis. CTC-supplementation was significantly (99% [95% CI=98-100%]) associated with increased tetracycline resistance compared to the control group (95% [95% CI=94-97%]). Copper supplementation was associated with decreased resistance to most of the antibiotics tested, including cephalosporins, over the treatment period. Overall, 91% of the E. coli isolates were multidrug resistant (MDR) (resistant to ≥3 antimicrobial classes). tetA and blaCMY-2 genes were positively associated (P<0.05) with MDR categorization, while tetB and pcoD were negatively associated with MDR. tetA and blaCMY-2 were positively associated with each other and in turn, these were negatively associated with both tetB and pcoD genes; which were also positively associated with one another. Copper minimum inhibitory concentration was not affected by copper supplementation or by pcoD gene carriage. CTC supplementation was significantly associated with increased susceptibilities of E. coli to copper (HR=7 [95% CI=2.5-19.5]) during treatment period. In conclusion, E. coli isolates from the nursery pigs exhibited high levels of antibiotic resistance, with diverse multi-resistant phenotypic profiles. The roles of copper supplementation in pig production, and pco-mediated copper resistance among E. coli in particular, need to be further explored since a strong negative association of pco with both tetA and blaCMY-2 points to opportunities for selecting a more innocuous resistance profile.

Effects of probiotic-supplemented diets on growth performance and intestinal immune characteristics of broiler chickens.

This study was conducted to investigate the effects of a probiotic product incorporating Lactobacillus fermentum and Saccharomyces cerevisiae on the growth performance and intestinal immune status in broiler chickens. A total of six hundred ninety-six 1-d-old male Cobb broilers were randomly allotted by BW in 1 of 4 treatments for 6-wk trial. The dietary treatments included the basal diet (NC), and the basal diets supplemented with an antibiotic (100 mg of chlortetracycline/kg of diet; PC), 0.1%, or 0.2% probiotic product (containing 1 × 10(7) cfu/g of Lactobacillus fermentum JS and 2 × 10(6) cfu/g of Saccharomyces cerevisiae). Each treatment had 6 replicates with 29 broilers each. The ADG and feed efficiency were improved (P < 0.05) in broilers fed the probiotic diet compared with NC, and were similar to the PC group during 1 to 21 d. However, there were no significant differences in growth performance of broilers during 22 to 42 d among different dietary treatments. Chicks fed probiotics had higher proportions of CD3+, CD4+, and CD8+ T-lymphocytes, whereas the antibiotic diet decreased the proportion of CD8+ T-lymphocytes in the foregut of broilers at 21 and 42 d compared with the NC group. No significant difference was observed in the mRNA expression level of chicken B-cell marker chB6 (Bu-1) in the foregut of chickens among different treatments. Probiotic-supplemented diets increased (P < 0.05) the mRNA expression levels of Toll-like receptor (TLR) 2 and TLR 4 at 21 d, and only the TLR2 mRNA level at 42 d in the foregut of chickens, but did not change (P > 0.05) TLR7 mRNA expression compared with NC or PC. There was no significant difference in the above TLR mRNA levels in the intestine of broilers between PC and NC. These results indicated that the probiotic product incorporating Lactobacillus fermentum and Saccharomyces cerevisiae could stimulate intestinal T-cell immune system without decreasing growth performance in broilers during 1 to 21 d.

Costs and outcomes over 36 years of patients with phenylketonuria who do and do not remain on a phenylalanine-restricted diet.

BACKGROUND: To quantify the costs and consequences of managing phenylketonuria (PKU) in the UK and to estimate the potential implications to the UK's National Health Service (NHS) of keeping patients on a phenylalanine-restricted diet for life. METHOD: A computer-based model was constructed depicting the management of PKU patients over the first 36 years of their life, derived from patients suffering from this metabolic disorder in The Health Improvement Network database (a nationally representative database of patients registered with general practitioners in the UK). The model was used to estimate the incidence of co-morbidities and the levels of healthcare resource use and corresponding costs over the 36 years. RESULTS: Patients who remained on a phenylalanine-restricted diet accounted for 38% of the cohort. Forty-seven per cent of patients discontinued their phenylalanine-restricted diet between 15 and 25 years of age. Of these, 73% remained off diet and 27% restarted a restricted diet at a mean 30 years of age. Fifteen per cent of the cohort had untreated PKU. Eleven per cent of patients who remained on a phenylalanine-restricted diet for 36 years received the optimum amount of prescribed amino acid supplements. Patients had a mean 12 general practitioner visits per year and one hospital outpatient visit annually, but phenylalanine levels were only measured once every 18 to 24 months. The mean NHS cost (at 2007/08 prices) of managing a PKU sufferer over the first 36 years of their life was estimated to range between £21 000 and £149 000, depending on the amount of prescribed nutrition they received. CONCLUSION: The findings suggest that the majority of patients with PKU were under-treated. The NHS cost of patient management should not be an obstacle to encouraging patients to remain on a restricted diet until further information becomes available about the long-term clinical impact of stopping such a diet. Nevertheless, patients require counselling and managed follow up regardless of the choices they make about their diet.

Effect of 2-mercaptoethanol and cysteine supplementation during in vitro maturation on the developmental competence of oocytes from hormone-stimulated lambs.

The objective was to determine the effect of 2-mercaptoethanol and cysteine on in vitro developmental competence of oocytes from lambs (4-8-week old) stimulated with eCG and pFSH. Oocytes were matured in medium (TCM199) with no supplement (Control group) or with 100 microM 2-mercaptoethanol and 600 microM cysteine (GSH group). Oocytes from adult sheep were also included (Adult group). The addition of 2-mercaptoethanol and cysteine did not improve nuclear maturation or microtubule configuration 12, 15, 18, or 24 h after placement in maturation medium. Sperm head decondensation and male pronucleus formation were evaluated at 6, 12, and 18 h after commencement of IVF; sperm decondensation appeared earlier in the GSH group (6h after the start of IVF). There were differences (P<0.05) between the Control group and the GSH and Adult groups for: fertilization rate at both 12 h (55.4, 77.0, and 80.6%, respectively) and 18 h (67.9, 86.9, and 88.7%); parthenogenesis rate at both 12 h (25.0, 10.8, and 5.6%) and 18 h (28.3, 9.8, and 4.5%); and polyspermy rate at 18 h (26.4, 4.9, and 5.7%). Blastocyst rate at 7d was higher in the GSH group than the Control group (23.9% vs. 14.9%, P<0.05), but both were lower (P<0.05) than the Adult group (38.3%). The addition of 2-mercaptoethanol and cysteine improved sperm decondensation and rates of fertilization and the blastocyst development to 7d, with no effect on blastocyst rate at 9 d.

Budesonide enema in pouchitis--a double-blind, double-dummy, controlled trial.

BACKGROUND: Pouchitis has been suggested to be a recurrence of ulcerative colitis in a colon-like mucosa. Topical steroids are a valid therapeutic alternative for distal forms of ulcerative colitis. AIM: To investigate the efficacy and tolerability of budesonide enema in the treatment of pouchitis compared with oral metronidazole. MATERIALS AND METHODS: Twenty-six patients with an active episode of pouchitis (defined as a pouchitis disease activity index score >or= 7) and no treatment during the previous month were randomized to receive either budesonide enema (2 mg/100 mL at bedtime) plus placebo tablets or oral metronidazole (0.5 g b.d.) plus placebo enema in a prospective, double-blind, double-dummy, 6-week, controlled trial. RESULTS: Based on the intention-to-treat principle, we detected a significant improvement in disease activity at the end of the first week with both drugs (P < 0.01). After that, improvement was moderated until stabilization at 4 weeks in both treatments. The per protocol analysis showed that both drugs had similar efficacy in terms of disease activity, clinical and endoscopic findings. Fifty-eight per cent and 50% of patients improved (decrease in pouchitis disease activity index >or= 3) with budesonide enema and metronidazole, respectively (odds ratio, 1.4; confidence interval, 0.2-8.9). Adverse effects were observed in 57% of patients given metronidazole and in 25% of patients given budesonide. CONCLUSIONS: Budesonide enemas are an alternative treatment for active pouchitis, with similar efficacy but better tolerability than oral metronidazole.

[Effects of musk glucoprotein on the function of rat polymorphonuclear leukocytes activated by IL-8 in vitro].

OBJECTIVE: To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on some functions of rat polymorphonuclear leukocytes activated by IL-8 in vitro. METHOD: An in vitro incubation system was used. Superoxide anion production was determined by cytochrome C reduction. beta-glucuronidase and lysozyme release was quantitated by enzyme reactions in which phenolph-thaleinglucuronic acid and Micrococcus Lysodeikticus were as the substrates, respectively. RESULTS: In comparison with control, musk-1 at concentration 1-100 micrograms.ml-1 can increase superoxide anion production by 91.7%-291%, and decrease beta-glucuronidase and lysozyme release by 2.2%-58.1% and 3.9%-39.8%, respectively. CONCLUSION: Inhibition of lysosomel enzyme release might be considered as one of mechanisms of antiinflammatory action of musk.

Binding of the maize cytosolic Hsp70 to calmodulin, and identification of calmodulin-binding site in Hsp70.

Using a gel-overlay technique of biotinylated calmodulin (CaM), we showed that maize cytosolic Hsp70 protein could bind to CaM in the presence of 1 mM CaCl2. The purified maize cytosolic Hsp70 inhibited the activity of CaM-dependent NADK in a concentration-dependent manner. A synthetic peptide, which possesses the 21 amino acid sequence, PRALRRLRTACERAKRTLSST, at positions 261-281 in maize cytosolic Hsp70, could associate with CaM in the presence of 1 mM calcium. The synthetic peptide inhibited CaM-dependent NADK activity and PDE activity. This indicates that the 21-amino acid sequence at positions 261-281 is the CaM-binding site. The binding of CaM to Hsp70 inhibited the ATPase activity of Hsp70. The possible regulator function of Hsp70 in cell signaling events in response to heat stress is discussed.

[Effects of musk glucoprotein on PAF production and cytosolic Ca2+ level in rat polymorphonuclear leukocytes in vitro].

OBJECTIVE: To investigate the effects of Musk glucoprotein on platelet activating factor (PAF) production and the concentration of cytosolic free Ca2+ in polymorphonuclear leukocytes of rat. METHODS: An in vitro incubation system was used, production of PAF and activity of acetyl transferase were measured by isotope incorporation, the concentration of cytosolic free Ca2+ was quantitated using the fluorescent Ca2+ indicator Fura-2. RESULTS: Musk-1 at concentration of 1-100 micrograms.ml-1 can significantly inhibit production of PAF, activity of acetyl transferase and the increase of cytosolic Ca2+ concentration in polymorphonuclear leukocytes of rat. CONCLUSION: Part of mechanisms underlying antiinflammatory action of Musk-1 is through inhibiting the synthesis of PAF and the increase of cytosolic Ca2+ level.

[The effect of integrin-like proteins on pollen germiantion and tube growth of Nicotiana tabacum].

The effects of antiserum against human VnR integrin and integrin antagonist GRGDSP peptides on Nicotiana tabacum pollen germination and tube growth both in in vitro and in semi-vivo conditions were studied. No obvious inhibitory effects on pollen germination and tube growth in vitro were observed when anti-VnR serum or GRGDSP peptides was added to BK culture medium, but the enhancement of pollen germination and tube growth in vitro promoted by calmodulin was depressed by adding anti-VnR serum or GRGDSP peptides to BK culture medium. In addition to that, pollen germination and tube growth on stigma, as well as tube growth in styles were also inhibited at some extent by treating stigma and microinjecting GRGDSP peptides or anti-VnR serum into styles. The role of integrin-like proteins in regulation of pollen germination and tube growth in situ was discussed.

Time course of nitric oxide synthase generation after gluten exposure in the rectal mucosa of gluten-sensitive patients.

BACKGROUND: Nitric oxide is thought to play an important role in modulating chronic inflammatory responses as well as in immune-mediated inflammation. We reproduced a gluten-mediated mucosal response in the rectum of celiac and control subjects in order to determine the role of inducible and constitutive nitric oxide synthases in the pathogenesis of this process. MATERIAL: Nine patients with confirmed celiac disease and five healthy controls underwent a long-term rectal gluten challenge (48 h) after an enema of 6 g of crude gluten, and constitutive and inducible nitric oxide synthase activity were determined in rectal biopsies. The histological localization of inducible nitric oxide synthase was determined by immunohistochemistry. RESULTS: Activity of both isoforms of nitric oxide synthase in control subjects did not change significantly after gluten instillation. In celiac patients, constitutive nitric oxide synthase on rectal mucosa also showed no significant changes after challenge with gluten. Inducible nitric oxide synthase isoform exhibited a modest increase 4 h after gluten instillation in celiac patients (mean increase 35% compared with baseline levels) but, 8 h after challenge, generation of iNO synthase was significantly higher: 54% more than pre-challenge production (P < 0.05) and higher than control values (P < 0.05). Inducible nitric oxide synthase staining was mostly localized in mononuclear cells of the epithelium and the lamina propria. After gluten instillation, the enhanced staining was mainly localized in subepithelial areas of the lamina propria. CONCLUSION: Our data suggest a role for nitric oxide, generated by inducible nitric oxide synthase, in the process of rectal mucosa injury by local gluten instillation in sensitized patients. We could not, however, determine if the role of nitric oxide in the ensuing injury of this gluten-induced immune inflammation model is a protective one, or merely a by-product generated by the activation of the inflammatory cells.

[Relationship between serum insulin, C-peptide in hypertension and syndrome differentiation-typing in TCM].

OBJECTIVE: To study the relationship between insulin-resistance (IR), hyperinsulinemia and TCM Syndrome Differentiation-typing. METHODS: The serum insulin, C-peptide level of the four Syndrome-types of hypertension (30 cases each type) and the control group (30 cases) were determined. RESULTS: The serum insulin level in hypertension patients were significantly higher than that of control group, and there were obvious difference among the four types of Syndrome. The following order was: The abundant phlegm-dampness type > exuberant Liver-Fire type > both Yin-Yang deficiency type > Yin deficiency and Yang-Excess type > control type. CONCLUSIONS: The Excess Syndrome was severe and deficiency Syndrome was mild in hyperinsulinemia. The pattern of change was in accordance with etiology and pathogenesis of TCM. It has the guiding significance to the clinical practice and research of TCM and integrated TCM-WM.