Cannabis sativa L. alleviates loperamide-induced constipation by modulating the composition of gut microbiota in mice.

MaZiRenWan (MZRW) is the most frequently used Traditional Chinese Medicine formula to treat chronic constipation, Cannabis sativa L. is regarded as a monarch drug in MZRW. However, the targets of Cannabis sativa L. that enhance colonic motility and improve constipation symptoms remain unknown. This study was designed to investigate the laxative effect and underlying mechanism of the water extract of Cannabis sativa L. (WECSL) using a loperamide-induced constipation mouse model. We found that WECSL treatment significantly improved intestinal motility and water-electrolyte metabolism, decreased inflammatory responses, prevented gut barrier damage, and relieved anxiety and depression in constipated mice. WECSL also structurally remodeled the composition of the gut microbiota and altered the abundance of bacteria related to inflammation, specifically Butyricicoccus and Parasutterella. Moreover, WECSL failed to relieve constipation symptoms following intestinal flora depletion, indicating that WECSL alleviates constipation symptoms depending on the gut microbiota. Our research provides a basis for WECSL to be further investigated in the treatment of constipation from the perspective of modern medicine.

[Contributions of medical practitioners of Xin'an area on acupuncture theory].

Contributions of Xin'an medical school and physicians to acupuncture theory were introduced in the article. Academic theories or characteristics of several physicians of Xin'an school such as YANG Xuan-cao, WU Kun, WANG Ji, WU Yi-ding, ZHENG Mei-jian and XU Chun-fu, et al were sorted out. Contributions of inheriting and illustrations on acupuncture theory were analyzed so as to expound its significance and value on modern acupucture clinic.

18α-Glycyrrhizin induces apoptosis and suppresses activation of rat hepatic stellate cells.

BACKGROUND: To investigate the potential mechanisms underlying the protective effects of 18α Glycyrrhizin (GL) on rat hepatic stellate cells (HSCs) and hepatocytes in vivo and in vitro. MATERIAL/METHODS: Sprague-Dawley (SD) rats were randomly divided into 5 groups: normal control group, liver fibrosis group, high-dose 18α GL group (25 mg/ kg/d), intermediate-dose 18α GL group (12.5 mg/kg/d) and low-dose 18α GL group (6.25 mg/ kg/d). The rat liver fibrosis model was induced by carbon tetrachloride (CCl4). The expressions of alpha-smooth muscle actin (αSMA) and NF-kappaB were determined by real-time PCR and immunohistochemistry. RESULTS: 18αGL dose-dependently inhibited the CCl4-induced liver fibrosis. There were significant differences in the mRNA and protein expressions of αSMA between the fibrosis group and 18α-GL treatment groups, suggesting that 18α GL can suppress the proliferation and activation of HSCs. Few HSCs were apoptotic in the portal area and fibrous septum in the liver fibrosis group. However, the double-color staining of a-SMA and TUNEL showed that 18α-GL treatment groups increased HSC apoptosis. NF-kappaB was mainly found in the nucleus in the fibrosis group, while cytoplasmic expression of NF-kappaB was noted in the 18αGL groups. In the in vitro experiments, 18α GL promoted the proliferation of hepatocytes, but inhibited that of HSCs. HSCs were arrested in the G2/M phase following 18α GL treatment and were largely apoptotic. CONCLUSIONS: 18α-GL can suppress the activation of HSCs and induce the apoptosis of HSCs by blocking the translocation of NF-kappaB into the nucleus, which plays an important role in the protective effect of 18α-GL on liver fibrosis.

[Immunoregulatory mechanisms of an optimal Chinese herbal monomer compound in mice with allergic rhinitis].

OBJECTIVE: To study the immunoregulatory effect of an optimal Chinese herbal monomer compound, which consists of three monomers, namely, icariin, baicalin and Astragalus saponin I, in a mouse model of allergic rhinitis. METHODS: A mouse model of allergic rhinitis was established by intraperitoneal injection of ovalbumin and aluminum hydroxide gel suspension. The splenic lymphocytes of the mice were separated, cultured in 96-well plates and divided into three groups: control group, concanavalin A group and compound group. Splenic lymphocyte proliferation was detected by cell counting kit-8 method at different time points. Cell cycle distribution was observed by flow cytometry (FCM) also at different time points. The changes of intracellular calcium concentration of splenic lymphocytes were measured by fluorescence microplate reader after the cells were incubated with fluorescence probe Fluo-3/AM. RESULTS: The Chinese herbal monomer compound could inhibit cell proliferation induced by concanavalin A (P<0.01). And the inhibition presented a time-effect relationship. With extending of the action time, the inhibition rate gradually increased and reached peak at the 48th hour. FCM test revealed the fact that concanavalin A could promote cells to enter into the mitosis by reducing the percentage of cells in G0/G1 phases while increasing the percentage of cells in S and G(2)/M phases. Compared with the concanavalin A, the compound could increase the percentage of cells in G(0)/G(1) phases and at the same time reduce the percentage of cells in S and G(2)/M phases at different time points, with the effect most significant at the 24th hour (P<0.05 or P<0.01). The results of the test taken by the fluorescence microplate reader revealed that the fluorescence value of the concanavalin A group increased with time in the previous 24 h while the compound could reduce this trend obviously, thus reduce the intracellular calcium concentration (P<0.01). CONCLUSION: The Chinese herbal monomer compound can inhibit the proliferation of cultured splenic lymphocytes of mice with allergic rhinitis. The effects of the compound of lowering intracellular calcium concentration and arresting cell cycle at G(0)/G(1) phases from entering into S and G(2)/M phases are responsible for its antiproliferation activity.

[Effects of Chinese herbal medicine Yiqi Zengmin formula on expression of glucose transporter 4 in skeletal muscle in type 2 diabetic rats].

OBJECTIVE: To investigate the effects of traditional Chinese compound recipe Yiqi Zengmin (YQZM) formula on expression of glucose transporter 4 (GLUT4) in skeletal muscle of rats with type 2 diabetes induced by high-fat diet combined with low-dose-streptozotocin injection. METHODS: Fifty male rats were randomly divided into two groups: model group fed with high-fat diet (n=40) and control group (n=10). After 4-week feed of high-fat diet, the rats of the model group were injected with streptozotocin at a dose of 35 mg/kg. Rats with plasma glucose over 16.67 mmol/L were randomly divided into 4 groups: diabetic model group, YQZM group, rosiglitazone group and losartan group and were treated with water, YQZM, rosiglitazone or losartan respectively by oral administration for 8 weeks. The expression of GLUT4 protein in the cytoplasm and the plasma membrane of isolated rats' skeletal muscles were detected by Western blotting. RESULTS: The expression of GLUT4 in the plasma membrane was more than that in the cytoplasm in skeletal muscle tissues of the normal rats, while the expression of GLUT-4 in the plasma membrane was less than that in the cytoplasm in skeletal muscle tissues of the type 2 diabetic rats. YQZM formula enhanced GLUT4 translocation from the cytoplasm to the plasma membrane in skeletal muscle tissues, similar to rosiglitazone. CONCLUSION: YQZM formula can enhance GLUT4 translocation from the cytoplasm to the plasma membrane in skeletal muscle tissues, and displays the insulin sensitization characteristic of rosiglitazone.

[Effects of Liangxue Tongyu Formula on brain edema and expressions of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in rats with intracerebral hemorrhage].

OBJECTIVE: To investigate the effects of Liangxue Tongyu Formula (LXTYF), a compound traditional Chinese herbal medicine, on brain edema in rats with intracerebral hemorrhage and to explore the mechanism. METHODS: Intracerebral hemorrhage was induced by using the intrastriatal autologous blood injection. Rats were randomized into sham-operated (SO) group, intracerebral hemorrhage (ICH) group and LXTYF group. Rats in the LXTYF group were intragastrically administered with LXTYF every day while the other two groups were given normal saline. Brain water content was determined at 24, 48, 72, and 120 h after intracerebral hemorrhage. Matrix metalloproteinase-9 (MMP-9) level, and MMP-9 and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA expressions in perihematoma area were detected by gelatin zymography and fluorescence quantitative real-time polymerase chain reaction at the four time points, respectively. RESULTS: Water content in the ICH group was highly elevated after intracerebral hemorrhage, and reached to the peak at 72 h. Compared with the ICH group, the LXTYF group had lower water contents at 48, 72 and 120 h after intracerebral hemorrhage (P<0.01). The difference in water content between the LXTYF and SO groups was significant only at 72 h (P<0.01). Although the pro-MMP-9 level and MMP-9 activity in the LXTYF and ICH groups were enhanced, they were still lower in the LXTYF group than in the ICH group (P<0.01 for 24, 48, 72 and 120 h, respectively). And there was no significant difference in them between the LXTYF group and the SO group at 120 h. Meanwhile, MMP-9 mRNA expressions were increased in the ICH and LXTYF groups, but the levels in the LXTYF group were significantly lower (P<0.01 for 48, 72 and 120 h, respectively) than those in the ICH group. Also, TIMP-1 mRNA expressions at 24, 48, 72 and 120 h after intracerebral hemorrhage were up-regulated in the LXTYF group, and there were significant differences in TIMP-1 expressions between the LXTYF group and ICH group after intracerebral hemorrhage (P<0.01). CONCLUSION: Liangxue Tongyu Formula ameliorates brain edema in rats after intracerebral hemorrhage by inhibiting MMP-9 expression and activity and up-regulating TIMP-1.

[Observation on therapeutic effect of acupoint application of Chinese medicine on bronchial asthma].

OBJECTIVE: To compare therapeutic effects of acupoint application of Chinese medicine and routine acupuncture on bronchial asthma. METHODS: One hundred cases were randomly divided into an observation group and a control group, 50 cases in each group. The observation group was treated with application of Chinese medicine at Dazhui (GV 14), Feishu (BL 13), Gaohuang (BL 43), Danzhong (CV 17), Qihai (CV 6) in the dog days; and the control group was treated with acupuncture at Zhongfu (LU 1), Tiantu (CV 22), Danzhong (CV 17), Dingchuan (EX-B 1), Fenglong (ST 40). The long-term therapeutic effects were observed. RESULTS: The total effective rate was 94.0% in the observation group and 76.0% in the control group with a significant difference between the two groups (P < 0.05). CONCLUSION: The therapeutic effect of acupoint application of Chinese medicine in the dog days on bronchial asthma is significantly superior to that of routine acupuncture.

[Effect of allyl glycoside extracted from Herba Rhodiolae on activity and proliferation of cultured Schwann cells under high glucose condition].

OBJECTIVE: To observe changes in activity and proliferation of cultured Schwann cells under the high glucose condition and effect of allyl glycoside extracted from Herba Rhodiolae (AG) on these changes for exploring the possible mechanism of diabetic peripheral nerve injury and the pharmacologic action of AG on them. METHODS: Adopting refined Brokes method, Schwann cells were isolated from the sciatic nerve tissue of newborn Wistar rats and purified. Their activity and proliferative capability were determined using XTT method and 3H-TdR incorporative method respectively. RESULTS: High glucose showed marked inhibitory effect on both activity and proliferative capability of Schwann cells, and the inhibition could be markedly improved by AG. CONCLUSION: Diabetic peripheral nerve injury is possibly related to inhibitory effect of high glucose on activity and proliferative capability of Schwann cells. Effect of AG in improving these inhibitory changes provides the experiment basis for clinical use of traditional Chinese medicine, Herba Rhodiolae.

[Effects of Pollen Typhae total flavones on mRNA expressions of peroxisome proliferator-activated receptors in 3T3-L1 adipocytes].

OBJECTIVE: To explore the effects of Pollen Typhae total flavones (PTF) on the mRNA expressions of peroxisome proliferator-activated receptors (PPARs) alpha,gamma, beta/delta so as to analyze its possible mechanism in improving the insulin sensitivity of 3T3-L1 adipocytes. METHODS: Adipocytes were treated with PTF, and expressions of PPARalpha, PPARgamma, PPARbeta/delta mRNAs relating to the adipocyte glucose and lipid metabolism were determined by reverse transcription polymerase chain reaction. RESULTS: PTF obviously up-regulated the expressions of PPARalpha and PPARgamma mRNAs, all showing significant differences as compared with those in the normal control group (P<0.01). CONCLUSION: With its function as an insulin sensitizer, PTF may enhance the PPARalpha and PPARgamma mRNA expressions in 3T3-L1 adipocytes.

[Effects of Yiqi Sanju Formula on non-alcoholic fatty liver disease: a randomized controlled trial].

OBJECTIVE: To observe the therapeutic effects of Yiqi Sanju Formula (YQSJF), a compound traditional Chinese herbal medicine, in treatment of non-alcoholic fatty liver disease (NAFLD). METHODS: Sixty-seven patients diagnosed with NAFLD were randomly divided into two groups: YQSJF-treated group (39 cases) and placebo group (28 cases). The NAFLD patients in the two groups were treated with YQSJF and placebo respectively for 3 months. Clinical symptoms, the CT ratio of liver-spleen, body mass index (BMI), waist circumference, homeostatic model assessment for insulin resistance (HOMA2-IR) and the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-alpha (TNF-alpha), high sensitivity C-reactive protein (hs-CRP), triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL) and low-density lipoprotein (LDL) were evaluated before and after treatment. RESULTS: After treatment, the clinical symptoms were improved and the levels of BMI, waist circumference, HOMA2-IR, ALT, AST, TG and TC were decreased significantly in the YQSJF-treated group (P<0.05). The CT ratio of liver-spleen in the YQSJF-treated group was increased significantly as compared with the placebo group (P<0.01).

[Pollen Typhae total flavones inhibit expression of interleukin-6 in C2C12 skeletal muscle cells cultured with palmitate].

OBJECTIVE: To observe the effects of Pollen Typhae total flavones (PTF) on expression of interleukin-6 (IL-6) mRNA and protein secretion in C2C12 cell strain of skeletal muscle cells cultured with palmitate, and to explore the mechanism of PTF in relieving insulin resistance (IR). METHODS: The IR of C2C12 cells was induced by co-culturing with palmitate. The C2C12 cells were divided into normal control group, untreated group, PDTC (a nuclear factor-kappaB inhibitor) treated group, rosiglitazone (ROS)-treated group, ROS+ PDTC-treated group, PTF-treated group and PTF+PDTC-treated group. Sixteen hours after culture, the transportation rate of glucose was observed by (3)H-deoxyglucose uptake method; IL-6 mRNA expression in C2C12 cells was assayed by real time polymerase chain reaction (Real-time PCR), and level of IL-6 protein secretion in culture supernatant was detected by enzyme linked immunosorbent assay. RESULTS: Compared with the normal control group, the transportation rate of glucose of cells in untreated group was decreased 30.43% after 16-hour palmitate culture, and was increased 32.39% in the PTF-treated group. Compared with the untreated group, the levels of IL-6 mRNA expression in cells and IL-6 protein secretion in supernatant were significantly decreased in the PTF-treated group (P<0.05). The levels of IL-6 mRNA expression in cells and IL-6 protein secretion in supernatant were increased in PTF+PDTC-treated group as compared with PFT-treated group (P<0.05). CONCLUSION: PTF can inhibit the IL-6 mRNA expression and IL-6 protein secretion via nuclear factor-kappaB pathway in C2C12 skeletal muscle cells, which may be one of its mechanisms in relieving inflammation conditions and insulin resistance in C2C12 skeletal muscle cells.

[A report on location of head acupoints by bone-length measurement in 100 persons].

OBJECTIVE: To compare the differences between bone-length measurements on human head and to probe proportion relation of the position of head points. METHODS: One hundred healthy adult volunteers, aged between 18-25 years, 50 males and 50 females, were randomly selected, and their height, body weight, the distances from Shenting (GV 24) to Toulinqi (GB 15), from Shenting to Touwei (ST 8), between bilateral Touwei (ST 8), between the two Mastoid, from Yintang (EX-HN 3) to front hairline, front hairline to Naohu (GV 17), Naohu to Fengfu (GV 16), Fengfu to the middle of rear hairline were measured respectively with standard measure instruments, and the proportion relation between bone-length measurement location and the point position were compared. RESULTS: There was a significant difference between the distance of bilateral Touwei and the distance of the two Wangu (P<0.05). The proportion relation of Shenting-Toulinqi, Shenting-Touwei, bilateral Touwei distances basically conformed to the bone-length measurement location, while in the proportion relation of the distances of Yintang-front hairline, front hairline-Naohu, Naohu-Fengfu, Fengfu-the middle of rear hairline and the bone-length measurement location there were some differences. CONCLUSION: Bilateral Touwei distance is not same as the bone-length measurements between two Mastoid, and Toulinqi can be located at the middle point of the connecting line of Shenting and Touwei. There is a certain deviation between marker location of Naohu, Fengfu and other points on body surface and the bone-length measurements.

[Study on changes of protein phosphorylation of p65, IkappaBalpha and IkappaBepsilon in lymphocytes of rats in progress of aging and interventional effect of Epimedium flavonoids].

OBJECTIVE: To study the characteristics of protein phosphorylation of p65, IkappaBalpha and IkappaBepsilon in lymphocytes of rats in the progress of aging and the the interventional effect of Epimedium flavonoids (EF). METHOD: We chose the lymphocytes derived from SD rat spleen. We divided the SD rats into five groups i. e. 4 months (4 m), 27 months (27 m), the 27 m EF treated group (27 m + EF), and we also observed the whole interventional effect of PDTC (a NF-kappaB inhibitor) on old rat groups (27 m PDTC, 27 m PDTC + EF ) when IkappaBepsilon, IkappaBalpha were detected. Through the western-blotting analyses, we studied the entire characteristics and distinctions of phosphorylation expression in molecules related to NF-kappaB signal transduction pathway p65, IkappaBepsilon and IkappaBalpha in lymphocytes across the age spectrum of rats in aging. We also observed the whole interventional effect of EF on lymphocytes of old rats. RESULT: With the increasing of age, the mean level of phosphorylation expressions of p65, IkappaBalpha and IkappaBepsilon in rat spleen lymphocytes decreased obviously, When inhibited NF-kappaB by PDTC, there was decreased evidently, while PDTC + EF can active NF-kappaB family and the above molecules were increased to a certain extent. CONCLUSION: The phosphorylation expressions of p65, IkappaBalpha and IkappaBepsilon in rat spleen lymphocytes were not enouphe, EF have a strong effect to upregulated the expression of them during aging.

[Effects of Astragalus polysaccharides on proliferation and differentiation of 3T3-L1 preadipocytes].

OBJECTIVE: To observe the effects of Astragalus polysaccharides (APS) on the proliferation and differentiation of 3T3-L1 preadipocytes and to elucidate its possible mechanism. METHODS: The proliferation of 3T3-L1 preadipocytes was detected by XTT method. Lipid droplets accumulated in cytoplasm of the differentiated preadipocytes were observed by using red O staining and quantified by colorimetry. The expressions of peroxisome proliferation activated receptor gamma (PPAR gamma) and CAAT/enhancer binding protein (C/EBP alpha) mRNAs and proteins were detected by real-time polymerase chain reaction (RT-PCR) and Western blotting respectively. RESULTS: APS at different concentrations (0.025-0.8 g/L) affected 3T3-L1 preadipocyte proliferation and differentiation dose-dependently. 3T3-L1 preadipocytes treated with 0.4 g/L APS had lots of lipid droplets in the cytoplasma, which were similar to cells treated with rosiglitazone (ROS). APS significantly increased the mRNA and protein expressions of PPAR gamma and C/EBP alpha (P<0.05, P<0.01, compared with the normal control group) in the course of 3T3-LI preadipocyte differentiation. CONCLUSION: APS can promote the proliferation of 3T3-L1 preadipocytes, enhance the accumulation of lipid drops, and increase the terminal differentiation of preadipocytes, which may be associated with its effects in increasing the expressions of PPAR gamma and C/EBP alpha mRNAs and proteins. The study suggests that APS has potential in the treatment of metabolic syndrome.

[Clinical research of Yiqi Sanju Formula in treating central obese men at high risk of metabolic syndrome].

OBJECTIVE: To observe the therapeutic effects of Yiqi Sanju Formula (YQSJF), a compound Chinese herbal medicine, on central obese men at high risk of metabolic syndrome (MS). METHODS: Compared with 30 healthy male volunteers, 45 central obese men were separated randomly into two groups and received the interventions with YQSJF and placebo respectively for 10 weeks. Baseline characteristics, insulin resistance, inflammation cytokines and plasminogen activator inhibitor-1 (PAI-1) and tissue plasminogen activator (t-PA) were evaluated before and after treatment. RESULTS: The score of homeostatic model assessment for insulin resistance (HOMA-IR) and the levels of C reactive protein (CRP), free fatty acid (FFA) and PAI-1 in obese men were higher than those in the control group, while t-PA was lower. After treatment, compared with placebo group, body mass index, waist, and waist-to-hip ratio were decreased significantly in subjects who received YQSJF (P<0.01). The score of HOMA-IR and the levels of CRP, FFA and PAI-1 were decreased significantly, and the level of t-PA was increased significantly (P<0.01). CONCLUSION: YQSJF can reduce obesity and insulin resistance in central obese men at high risk of MS and improve inflammation and fibrinolysis, which indicates that it can reduce the risk of atherosclerosis.

[Comparison of gene expression profiles between rats of Shen deficiency syndrome and Shen-yang deficiency syndrome differentiated according to therapeutic efficacy of drugs used].

OBJECTIVE: To explore similarity and difference of connotation between Shen deficiency syndrome (SDS) and Shen-yang deficiency syndrome (SYDS) on the molecular level. METHODS: The senescent SD rats and corticosterone-treated rats were adopted for models of SDS and SYDS respectively, their syndrome was differentiated according to the therapeutic efficacy of treatment with epimedium flavonoids (EF). The gene expression profiles of hypothalamas, pituitary, adrenal gland and lymphocytes (HPAT axis) were detected before and after EF treatment using gene chip provided by Affymetrix company. RESULTS: As compared with the young rats, the ageing rats and corticosterone-treated rats showed a significant down-regulation in highly consistent pattern, of various neurotransmitters of HPAT axis firstly, followed with that of growth and sex hormone related genes. EF could reverse the above genes expression in both models, and for SYDS model rats, it could also significantly up-regulate the gene expressions of heat shock protein, cytochrome P450 and thyroid stimulating hormone (TSH). CONCLUSION: Both SDS and SYDS model rats show connotation of Shen deficiency, and the substantial base of Shen-yang deficiency syndrome resides in the process of oxidative phosphorylation of energy metabolism accelerated by thyroid hormone.

[Effects of Pollen Typhae total flavone on glucose and lipid metabolism in 3T3-L1 adipocytes].

OBJECTIVE: To observe the effects of Pollen Typhae total flavone (PTF) on glucose and lipid metabolism in 3T3-L1 adipocytes. METHODS: The content of glucose which disappeared from the culture medium after incubation with drugs for 24 hours was determined as glucose consumption of the cells. The activity of cells was detected by XTT method. The transport of glucose was observed by (3)H-glucose uptake method. The efflux of free fatty acid (FFA) from adipocytes was observed by the concentration of FFA in the culture medium. RESULTS: The glucose concentration in culture medium was significantly decreased with a concentration-dependent effect, when PTF concentrations were from 0.025 g/L to 0.4 g/L. The toxic effect on cells appeared while PTF concentration was 0.4 g/L, and the MTT value decreased. PTF also significantly increased glucose transportation in the 3T3-L1 adipocytes as rosiglitazone (ROS) did. At the same time, FFA concentration in culture medium was significantly decreased as compared to the normal control group, while ROS-treated group did not show any difference. CONCLUSION: PTF can increase insulin sensitivity by increasing glucose transportation and consumption in the 3T3-L1 adipocytes as well as decreasing the FFA efflux from the cells.

[Exploration on molecular mechanism of Epimediun flavonoids in regulating adrenocortical regeneration in rats with inhibited hypothalamic-pituitary-adrenal axis using oligonucleotide microarrays].

UNLABELLED: To investigate the regulatory effects of epimedium flavonoids (EF) on adrenocortical regeneration in rats with inhibited hypothalamic-pituitary-adrenal (HPA) axis. METHODS: Cell distribution in cell cycle and cell apoptotic rate were measured with PI stain and flow-cytometry; apoptosis cells were showed by in situ terminal-deoxynucleotidyl transferase-mediated deoxy-uridine triphosphate-fluorescene nick end labeling assay (TUNEL), and the genome-wide gene mRNA expression was detected by oligonucleotide microarrays. RESULTS: Compared to the normal control, adrenal cells isolated from the HPA axis inhibited model group were arrested in Go/GI phase, and showed a higher apoptotic rate (P < 0.05). After treated with EF, cells in G0/G1 phase decreased and those in G2/M phase increased (P < 0.01), and the elevated apoptotic rate reduced significantly (P < 0.05). TUNEL assay showed the number of apoptotic cells per section was 4.67 1.53 in the normal control group, 70.67 +/- 9.29 in the model group, and 18.67 +/- 7.64 in the EF-treated group respectively (n=3). Gene expressions in adrenal were mostly restrained in the model group, including 7 cytocycle promoting genes, including V-ras, V-jun, etc., while after treatment with EF, 6 cytocycle promoting genes, 1 anti-apoptotic gene, and genes that closely related with adrenocortical regeneration as IGF-II and FGF7 and their receptors, as well as 7 steroid biosynthesis participated genes were all up-regulated. Conclusion EF can accelerate adrenocortical cell proliferation, inhibit its apoptosis, and promote steroid biosynthesis so as to enhance adrenocortical regeneration in HPA axis inhibited rats, which may contribute to the beneficial effects of EF in protecting adrenocortical function during glucocorticoid withdrawal.

Deguelin regulates nuclear pore complex proteins Nup98 and Nup88 in U937 cells in vitro.

AIM: To investigate the anticancer effects and the molecular mechanisms of deguelin on human U937 leukemia cells, and to explore the underlying mechanism regulating nucleoporin 98 (Nup98) and nucleoporin 88 (Nup88) in vitro. METHODS: The effects of deguelin on the growth of U937 cells were studied by MTT assay. The effect of deguelin on the cell cycle of U937 cells was studied by using a propidium iodide method. The localization of the nuclear pore complex proteins Nup98 and Nup88 was investigated by using immunofluorescence and immunoelectron microscopy. The expression of Nup98 and Nup88 in U937 cells was investigated by using flow cytometry and Western blot. RESULTS: The proliferation of U937 cells was inhibited in the deguelin-treated group, with a 24-h IC(50) value of 21.61 nmol/L and a 36-h IC(50) value of 17.07 nmol/L. U937 cells treated with deguelin had reduced percentages of cells in the G(0)/G(1) phase, whereas cells accumulated in the S and G(2)/M phases. Nup88 and Nup98 were found on both the nuclear and cytoplasmic sides of the U937 cells by using immunofluorescence and immunoelectron microscopy. The expression of Nup98 was upregulated and that of the Nup88 protein was downregulated in U937 cells treated with deguelin. CONCLUSION: Deguelin is able to inhibit the proliferation of U937 cells by regulating the cell cycle such that cells are arrested at the S and G(2)/M phases, so that the proportion of cells in the G(0)/G(1) phase decreases. The antitumor effects of deguelin are related to upregulating the expression of Nup98 and downregulating the expression of Nup88 protein in U937 cells.

[Study on the differentially expressed genes in bone tissue of ovariectomized rats after intervention with Chinese herbs].

OBJECTIVE: To explore the therapeutic effect and mechanism of jiangu granule (JGG) on postmenopausal osteoporosis. METHODS: Differential display reverse transcription PCR (DDRT-PCR) was used to detect the differentially expressed genes in bone tissues associated with therapeutic effect of JGG. RESULTS: Ten differentially expressed gene fragments were found, 9 of them, after cloning, sequencing and BLAST searching, were proved to be matched with highly homologous sequences in Genebank. Among them, two were known proteins: hyaluronan-mediated motility receptor (RHAMM) and ATPase, Na+, K+ transporting beta 3 polypeptide (ATP1b3). It was confirmed by semi-quantitative RT-PCR that these two gene fragments could be down- and up-regulated by JGG respectively. CONCLUSION: The therapeutic effect of JGG in treating postmenopausal osteoporosis might be related with its regulation on the two kinds of protein, RHAMM and ATP1b3.