The chemical constituents of Poria cocos were studied by means of silica gel, ODS column chromatography, Sephadex LH-20 and preparative HPLC. Thirteen compounds were isolated from this plant. By analysis of the ESI-MS and NMR data, the structures of these compounds were determined as tumulosic acid (1), dehydrotumulosic acid (2), 3beta, 5alpha-dihydroxy-ergosta-7, 22-dien-6-one (3), 3beta, 5alpha, 9alpha-trihydroxy-ergosta-7, 22-diene -6-one (4), ergosta-7, 22-diene-3-one (5), 6, 9-epoxy-ergosta-7,22-diene-3-ol (6), ergosta-4,22-diene-3-one (7), 3beta, 5alpha, 6beta-trihydroxyl-ergosta-7,22-diene (8), ergosta-5, 6-epoxy-7,22-dien-3-ol (9), beta-sitosterol (10), ribitol (11), mannitol (12), and oleanic acid 3-O-acetate (13), respectively. Compounds 3-13 were isolated from the P. cocos for the first time.
Drugs, Chinese Herbal ; chemistry ; Organic Chemicals ; analysis ; Poria ; chemistry

OBJECTIVE Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disease with a high morbidity around 1/1000-1/400, characterized by progressive enlargement of fluid-filled cysts derived from renal tubular epithelial cells. Massive cysts gradually compress renal parenchyma destroying normal renal structures and compromising renal functions. Unfortunately, it will cause end-stage renal disease in most of the patients but without effective therapy now, who have to live on hemodialysis or kidney transplantation. Based on this present situation, it is of great significance to find early intervention to inhibit renal cyst development. The projective of this study was to investigate whether Ganoderma triterpenes (GT) can inhibit renal cyst development and study the related mechanism. METHODS and RESULTS First, we used MDCK cyst model, cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells. GT inhibited MDCK cyst formation significantly, and inhibited cyst enlargement dose-dependently proving GT cyst inhibition in vitro. Then we used an embryonic kidney cyst model, wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-cAMP. GT inhibited embryonic kidney cyst development significantly in a dose-dependent and reversible manner proving GT cyst inhibition at organ level. Furthermore, we used two ADPKD mouse models with severe cystic kidney disease phenotypes. GT dramatically inhibited renal cyst development, decreased ADPKD mouse kidney volume and the cyst index inside proving GT cyst inhibition in vivo. By Western blot, we proved GT down-regulated Ras/MAPK signal pathway without detectable effect on mTOR signal pathway both in MDCK cells and two ADPKD mouse kidneys. CONCLUSION GT retard renal cyst development both in vitro and in vivo significantly. The related mechanisms were involved in GT promoting renal tubular epithelial cell differentiation, down-regulating intracellular cAMP level and Ras/MAPK signal pathway.

OBJECTIVE To investigate the protective effect and potential mechanism of desmethylbellidifolin (DMB) in chronic alcoholic fatty liver disease. METHODS C57BL/6 mice were randomly divided into five groups. Control, meta?doxine and DMB group (high dose and low dose) mice were fed with Lieber-DeCarli liquid diet containing 5％alcohol for six weeks. Pair-fed group mice were fed with a liquid diet containing the same calories. After treatment, serum GOT, GPT, TG and hepatic T-CHO, TG, GSH, GSH-Px, SOD and CAT levels were measured. Ectopic liver lipid deposition was determined by oil red O and hematoxylin-eosin (HE) staining. Lipid metabolism and autophagy related genes expression were determined by real-time PCR and Western blotting. Electron microscope and laser scanning confocal microscope were used to detect autophagosome and autophagy flux. RESULTS DMB treatment markedly reduced serum TG, GOT and GPT levels in alcohol-induced mice, as well as hepatic levels of T-CHO, TG and MDA, while increased the GSH, GSH-Px, SOD and CAT levels in the liver. Oil red O and HE staining showed that the alcohol-induced lipid accumulation and hepatocyte morphology changes were significantly improved by DMB treatment. Mecha?nistically, the expression of stearoyl-CoA desaturase 1 and fatty acid synthase were significantly decreased, while lipoly?sis related hormone-sensitive lipase was elevated in mouse liver by DMB treatment. In addition, DMB could inhibit the phosphorylation of Akt and mTORC1, and activate autophagy process by inducing autophagy related genes expression, such as LC3, ATG5 and ATG7. Moreover, treatment with DMB notably increased the number of autolysosome and promote the autophagy flux, which may therefore induce the lipolysis and oxidation of lipids and prevent the alcohol-induced excessive lipid accumulation in the liver. CONCLUSION DMB exerts a protective role in alcoholic fatty liver dis?ease by regulating the Akt-mTORC1 pathway mediated autophagy activation.

OBJECTIVETo compare therapeutic effects of acupuncture and Western medicine for promoting ovalation on endocrine dysfunctional infertility.

METHODSTwo hundred and forty cases of infertility were randomly divided into an acupuncture group (n = 160) and a Western medicine group (n = 80). They were treated with acupuncture and clomiphene respectively and their therapeutic effects were compared.

RESULTSThe pregnancy rate was 65.0% in the acupuncture group and 45.0% in the Western medicine group with a significant difference between the two groups (P < 0.05).

CONCLUSIONAcupuncture can cure endocrine dysfunctional infertility.

Acupuncture Points ; Acupuncture Therapy ; Clomiphene ; Humans ; Infertility ; Medicine, Chinese Traditional ; Pregnancy Rate

Carcinoma, Renal Cell ; Humans ; Kidney Neoplasms ; Nomograms ; Prognosis ; Progression-Free Survival ; Retrospective Studies

OBJECTIVETo examine modulations caused by cyclooxygenase-2 (COX-2) inhibitors on altered microenvironments and overbalanced neurotransmitters in pilocarpine-induced epileptic status rats and to investigate possible mechanisms.

METHODSCelecoxib (a COX-2 inhibitor) was administered 45 min prior to pilocarpine administration. The effects of COX-2 inhibitors on mIPSCs (miniature GABAergic inhibitory postsynaptic currents) of CA3 pyramidal cells in the hippocampus were recorded. Expressions of COX-2, c-Fos, newly generated neurons, and activated microgliosis were analyzed by immunohistochemistry, and expressions of alpha-subunit of gamma-amino butyric acid (GABA(A)) receptors and mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MAPK/ERK) activity were detected by Western blotting.

RESULTSPretreatment with celecoxib showed protection against pilocarpine-induced seizures. Celecoxib prevented microglia activation in the hilus and inhibited the abnormal neurogenesis and astrogliosis in the hippocampus by inhibiting MAPK/ERK activity and c-Fos transcription. Celecoxib also up-regulated the expression of GABA(A) receptors. NS-398 (N-2-cyclohexyloxy-4-nitrophenyl-methanesulfonamide), another COX-2 inhibitor, enhanced the frequency and decay time of mIPSCs.

CONCLUSIONThe COX-2 inhibitor celecoxib decreased neuronal excitability and prevented epileptogenesis in pilocarpine-induced status epilepticus rats. Celecoxib regulates synaptic reorganization by inhibiting astrogliosis and ectopic neurogenesis by attenuating MAPK/ERK signal activity, mediated by a GABAergic mechanism.

Animals ; Blotting, Western ; Celecoxib ; Cyclooxygenase 2 ; metabolism ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Disease Models, Animal ; Fibrocystic Breast Disease ; metabolism ; Hippocampus ; drug effects ; enzymology ; pathology ; Immunohistochemistry ; MAP Kinase Signaling System ; drug effects ; Male ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Nitrobenzenes ; pharmacology ; Pilocarpine ; Proto-Oncogene Proteins c-fos ; metabolism ; Pyrazoles ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, GABA-A ; biosynthesis ; Status Epilepticus ; chemically induced ; enzymology ; pathology ; Sulfonamides ; pharmacology ; Synapses ; drug effects ; pathology

To investigate the effect of treadmill running on the ability of learning in young rats, male Sprague-Dawley rats (5 weeks of age) were used for the experiment. Animals were randomly divided into the control and running groups (n=15 in each group). The rats in running group were made run on a motor-driven treadmill for 1 week at a speed of 2 m/min for the first 5 min, at a speed of 5 m/min for the next 5 min, then at a speed of 8 m/min for the last 20 min. Then the Morris water maze was used to observe learning and memory ability of rats in both groups. The tests consisted of place navigation and spatial probe test. We found that, in place navigation training, the latency of rats in running group was less than that in control group (P<0.05); and from the third training session on, there was significant difference between the rats in control and running groups in swimming velocity (P<0.01); furthermore, it was observed that the rats in running group had stronger motivation and more exact orientation in searching for platform, which could be indicated by the index of turn angle and angular velocity. In spatial probe test, there was no significant difference between the two groups in swimming velocity, percentage of swimming distance and frequency of crossing platform in D quadrant, where the platform situated (P>0.05). These findings suggest that low speed treadmill running can enhance the ability of learning in young rats.
Age Factors ; Animals ; Male ; Maze Learning ; physiology ; Memory ; physiology ; Physical Conditioning, Animal ; physiology ; Rats ; Rats, Sprague-Dawley ; Spatial Behavior ; physiology ; Swimming ; physiology

Objective·To discover lead compounds with 6,11-dihydro-5H-benzo[a]carbazole as core scaffold that can inhibit the proliferation of cervical cancer cells. Methods?·?A series of 6,11-dihydro-5H-benzo[a]carbazole derivatives and analogs were synthesized using Fischer indole synthesis method, and their anticancer activity against HeLa cells was tested in vitro by CCK8 test. Results?·?2-Methoxy-6,11-dihydro-5H-benzo[a]carbazole and 8-chloro-2-methoxy-6,11-dihydro-5H-benzo[a]carbazole could significantly inhibit the proliferation of HeLa cells with the half maximal inhibitory concentration ( IC50) values of 9.61?μmol/L and 16.52?μmol/L, respectively. Conclusion?·?Two objective lead compounds were found. Among 6,11-dihydro-5H-benzo[a]carbazole derivatives, compounds with methoxy group at the C-2 position of the core scaffold show better activity against proliferation of cervical cancer cells.

Objective The neuronal ceroid lipofuscinosis (CLN are a group of severe lysosomal storage diseases.The patients present with clinically and genetically heterogeneous neurodegenerative disorders.This study aims to investigate the clinical characteristics and the gene mutations of a rare Chinese family with 3 siblings affected by CLN7.Methods The proband,a 5-year-old girl,visited us because of intermittently seizures and mental retardation for 2 years and a half in December,2015.Clinical investigation,brain magnetic resonance imaging(MRI),biochemical and the gene analysis were performed for the etiological study.Results The proband had seizures at the age of 2 and a half years,with the progressive motor deterioration,speech disturbance,mental regression and vision loss.Her brain MRI showed diffusive cerebral atrophy.The blood aminoacids,acylcarnitine and urine organic acid profiles were normal.Lysosomal palmitoyl protein thioesterase and tripeptidyl peptidase activities of peripheral leukocytes were normal.A compound heterozygous mutation of c.1351-1G ＞ A and c.300T ＞ G was detected on her MFSD8 gene,supporting the diagnosis of CLN7.Both of the 2 mutations were novel.Each of her parents carried one of the mutations.Two brothers of the proband had similar clinical process.Her elder brother died at the age of 7 due to severe encephalopathy of unknown etiology.The younger brother showed dyskinesia from the age of 2 years and seizures from the age of 4 years.A compound heterozygous mutation on MFSD8 gene,c.1351-1G ＞ A and c.300T ＞ G,was found from the younger brother,as same as the proband.Conclusions CLN7 is a rare disorder of CLN.In this study,the diagnosis of the 3 siblings with similar clinical process were much delayed.Gene analysis was key for the diagnosis.Two novel mutations were found on MFSD8 of the family.There is still no effective treatment for neurol ceroid lipofuscinosis.The prognosis is poor.Based on the mutation diagnosis,prenatal diagnosis for the next sibling is possible to the prevention of the disease.

OBJECTIVETo investigate the cellular immune regulation of the long-term intervention of moxa smoke.

METHODSThirty-two Wistar rats were randomly divided into a blank group, a low concentration group, a medium concentration group and a high concentration group, 8 cases in each group. In addition to the blank group, rats in the other groups were exposed to the corresponding concentration moxa smoke for 20 min every day, the T lymphocyte subsets and proportion of the CD4+ CD25+ Treg in CD4+ T cells in peripheral blood were tested by flow cytometry after 6 months.

RESULTSCompared with the blank group, the proportions of CD3+ CD4+, CD3+ CD8+ T cells and CD3+ CD4/CD3+ CD8+ in the other 3 moxa smoke groups were not significantly different (P > 0.05), while the proportions of the CD4+ CD25+ Treg in CD4+ T cells were significantly lower (P < 0.05), but no statistically significant differences among those 3 moxa smoke intervention groups (P > 0.05).

CONCLUSIONLong-term moxa smoke intervention has no significant effect on the proportions of CD3+ CD4+, CD3+ CD8+ T cells and CD3+ CD4+/CD3+ CD8+, but it can decrease the proportions of the CD4+ CD25+ Treg in CD4+ T cells in peripheral blood of rats. The way produced by pretreatment with moxa smoke may play immunomodulatory effect.

Animals ; Lymphocyte Count ; Male ; Moxibustion ; Rats ; Rats, Wistar ; Smoke ; analysis ; T-Lymphocyte Subsets ; drug effects ; immunology ; T-Lymphocytes, Regulatory ; drug effects ; immunology ; Time Factors