A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18β-glycyrrhetinic acid(1), 3-oxo-15β-hydroxy-18β-glycyrrhetinic acid (2), 7β,15α-dihydroxy-18β-glycyrrhetinic acid (3), 3-oxo-7β, 15α-dihydroxy-18β-glycyrrhetinic acid (4), 7β-hydroxy-18β-glycyrrhetinic acid(5) and 15α-hydroxy-18β-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation
Biotransformation ; Cunninghamella ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; metabolism ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Apoptosis of abnormal oocytes is essential for defective oocyte elimination during prepubertal ovary development, and the ubiquitin system regulates the cell apoptosis via the degradation of specific proteins. Ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is a component of the ubiquitin system, and the UCH-L1-dependent apoptosis is important for spermatogenesis. In the present study, the change in the number of follicles and the expression of UCH-L1 in oocytes were determined in prepubertal mouse ovaries by immunohistochemical techniques. A significant decrease in the follicular pool was found in prepubertal mouse ovaries during the period of day 21 to day 28 after birth, and accordingly, the UCH-L1 protein expression was increased, to some degree in association with Jun activation domain-binding protein 1 (Jab1) and cyclin-dependent kinase inhibitor p27(Kipl). The increased UCH-L1 protein, together with the corresponding changes of Jab1 was detected in morphologically abnormal oocytes of prepubertal ovaries. Through the immunofluorescent colocalization, UCH-L1 was shown concentrating in abnormal oocytes, and a parallel change in Jab1 was also seen. The affinity analysis confirmed the interaction between UCH-L1 and Jab1 in ovaries. These results suggest that UCH-L1 plays an important role, possibly in association with Jab1 and p27(Kipl), in selective elimination of abnormal oocytes during mouse prepubertal development.
Animals ; Apoptosis ; Female ; Mice ; Oocytes ; cytology ; Ovary ; enzymology ; Ubiquitin Thiolesterase ; metabolism

OBJECTIVETo study the alkaloidial constituents of the leaves of uncaria hirsuta.

METHODSome chromatographic methods were applied to isolate pure compounds and their structures were elucidated by spectroscopic methods.

RESULTEleven compounds were isolated and identified as 19-epi-3-iso-ajmalicine (1), 3-isoajmalicine (2), harman (3), mitraphylline (4), isomitraphylline (5), isorhynchophylline (6), corynoxine (7), rhynchophylline (8), isomitraphyllic acid (9), uncarine A (10) and uncarine B (11).

CONCLUSIONCompounds 1-9 were firstly isolated from this plant.

Alkaloids ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Plant Leaves ; chemistry ; Uncaria ; chemistry

OBJECTIVETo investigate the effect of air pollution on stroke mortality in Tianjin, China, and to provide basis for stroke control and prevention.

METHODSTotal data of mortality surveillance were collected by Tianjin Centers for Disease Control and Prevention. Meteorological data and atmospheric pollution data were from Tianjin Meteorological Bureau and Tianjin Environmental Monitoring Center, respectively. Generalized additive Poisson regression model was used in time-series analysis on the relationship between air pollution and stroke mortality in Tianjin. Single-pollutant analysis and multi-pollutant analysis were performed after adjustment for confounding factors such as meteorological factors, long-term trend of death, "days of the week" effect and population.

RESULTSThe crude death rates of stroke in Tianjin were from 136.67 in 2001 to 160.01/100000 in 2009, with an escalating trend (P = 0.000), while the standardized mortality ratios of stroke in Tianjin were from 138.36 to 99.14/100000, with a declining trend (P = 0.000). An increase of 10 µg/m³ in daily average concentrations of atmospheric SO₂, NO₂ and PM₁₀ led to 1.0105 (95%CI: 1.0060 ∼ 1.0153), 1.0197 (95%CI: 1.0149 ∼ 1.0246) and 1.0064 (95%CI: 1.0052 ∼ 1.0077), respectively, in relative risks of stroke mortality. SO₂ effect peaked after 1-day exposure, while NO₂ and PM₁₀ effects did within 1 day.

CONCLUSIONAir pollution in Tianjin may increase the risk of stroke mortality in the population and induce acute onset of stroke. It is necessary to carry out air pollution control and allocate health resources rationally to reduce the hazard of stroke mortality.

Air Pollutants ; analysis ; Air Pollution ; analysis ; China ; epidemiology ; Environmental Monitoring ; Humans ; Models, Theoretical ; Particulate Matter ; analysis ; Poisson Distribution ; Stroke ; epidemiology ; mortality ; Survival Rate ; Time Factors

OBJECTIVETo investigate the development of nitric oxide synthase (NOS)-positive neurons in the frontal lobe of the cerebrum of human fetus in midanaphase.

METHODSThe positive expression of the NOS-positive neurons in the frontal lobe of cerebrum of human fetus was observed by immunohistochemistry.

RESULTSBy the 7th to 8th month of gestation, NOS-positive neurons in the cortical plate of frontal lobe demonstrated themselves inequality of sizes and morphological difference in the deeper layers with interspersed distribution and increased NOS response, and the distribution of beaded nerve fiber was observed between neurons of cerebral tissues. By the 9th to 10th month of gestation, NOS-positive neurons in the deeper layers of cortical plate of frontal lobe developed slightly in size of the cell body with richer cytoplast, full shape and deeper dyeing and extrusive beaded nerve fibers, and the NOS-positive neurons scattered in the shallow layer of cortical plate presented with round or oval shape. The nucleus developed bigger but with sparse cytoplasm and clear nerve process.

CONCLUSIONNOS-positive neurons in the deeper layer of cortical plate of lobus frontal consist of largely network of neural system and produce micro-environment with higher concentration of NO, which favors the differentiation, proliferation, migration, and development of various neurons.

Cerebrum ; cytology ; embryology ; Female ; Fetus ; cytology ; enzymology ; Frontal Lobe ; cytology ; embryology ; Humans ; Immunohistochemistry ; Neurons ; cytology ; enzymology ; Nitric Oxide Synthase Type I ; metabolism ; Pregnancy

OBJECTIVETo elucidate the relationship between surgical time and postoperative complications in senile patients with hip fractures, and try to find out other factors which are related to these complications.

METHODSSixty-two patients, 28 males aged from 65 to 72 years with a mean age of 76.3 years and 34 females aged from 65 to 95 years with a mean age of 78.1 years, who had undergone orthopedic surgery because of hip fractures, were enrolled in a retrospective cohort study. The surgical time and pattern, the type of fracture, preoperative comorbidities, American Society of Anesthesiologists (ASA) score and the volume of blood transfusion during operation were obtained from these patients who were followed up by telephone calls for postoperative complications. All the patients were followed up at least for 1 year and were divided into subgroups according to their clinical characteristics and the results were analyzed by the Statistical Analysis System software.

RESULTSThere was no significant difference in the morbidity of postoperative complications with the gender, age, surgical time and pattern, or ASA score. There was significant difference in the morbidity of postoperative complications related to preoperative comorbidities and the volume of blood transfusion. There was a significant causality between preoperative comorbidities and postoperative complications. The morbidity of postoperative complications was 1.651 times higher in patients with preoperative comorbidities than those without.

CONCLUSIONSThere is no relationship between the surgical time and postoperative complications in senile patients who received surgery for hip fracture within 1 year. No correlation is found between the postoperative complications and gender, age, type of fracture, surgical pattern, ASA score and the volume of blood transfusion. Preoperative comorbidities are an independent predictor for postoperative complications.

Aged ; Aged, 80 and over ; Cohort Studies ; Female ; Hip Fractures ; surgery ; Humans ; Logistic Models ; Male ; Morbidity ; Postoperative Complications ; epidemiology ; Retrospective Studies ; Time Factors

OBJECTIVETo study the interactive effect of job stress and genetic susceptibility (or gene polymorphism) on hypertension.

METHODSA cross-sectional epidemiological study was conducted in 452 workers from a thermal power plant in China. Extrinsic effort, occupational reward, and over-commitment were measured. Hypertensive patients were defined by three phases of screening, reexamination, and final diagnosis. beta2-AR genotypes and allele frequencies at amino acid positions 16 (beta2-AR-16: Arg-->Gly) and 27 (beta2-AR-27: Gln-->Glu) were identified by PCR-RFLP.

RESULTSJob stress was related with the prevalence of hypertension in males (P < 0.05), whereas no significant relationship was found in females (P > 0.05). Differences in genotypes and allele frequencies of the beta2-AR-16 were statistically significant between the hypertension and control groups (P < 0.05), whereas those of beta2-AR-27 were not (P > 0.05). The prevalence of hypertension was higher in individuals carrying Gly16 allele than in those carrying Arg16 allele of the high job stress group (P < 0.01 or 0.05).

CONCLUSIONHigh job stress and polymorphism of beta2-AR-16 have an interactive effect on the prevalence of hypertension in male workers.

Adult ; China ; epidemiology ; Employment ; psychology ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Hypertension ; epidemiology ; genetics ; Male ; Polymorphism, Genetic ; Power Plants ; Prevalence ; Receptors, Adrenergic, beta-2 ; genetics ; Stress, Psychological ; complications ; Young Adult

OBJECTIVETo investigate the growth and development of nitric oxide synthase (NOS)-positive neurons in the cerebellum of human fetus in the midanaphase.

METHODThe positive expression of the NOS-positive neurons in the cerebellum of midanaphase human fetus was observed by immunohistochemistry.

RESULTSBy the sixth to seventh month of gestation, NOS-positive neurons were seen in the ependymal layer of the cerebellum. The nucleus was oval-shaped and the neurons had short and small processes. By the eighth to ninth month, NOS-positive neurons were found in the central layer of the cerebellum and the nucleus was round-, oval-, or fusiform-shaped; meanwhile, the neurons grew larger in size with richer cytoplast and heavier staining. The beaded nerve fibers reached the marginal layer and the layer became thickened on the tenth month, which generally was composed of 5 to 6 layers of NOS-positive neurons that were tightly aligned. Some NOS-positive neurons were in smaller size with the cell body and the nerve fibers grew well.

CONCLUSIONNitric oxide generated by NOS of the NOS-positive neurons in the cerebellum plays an important role in the differentiation, proliferation, and migration of neurons and gliacytes.

Cerebellar Cortex ; Fetus ; physiology ; Humans ; Immunohistochemistry ; Nerve Fibers ; Neurons ; cytology ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Nitric Oxide Synthase Type I

Objective:To explore the role of macrophage polarization on pericyte-to-myofibroblast transition and renal allograft fibrosis after kidney transplantation(KT).Methods:Allograft tissues were harvestedfrom recipients with chronic allograft dysfunction(CGD)and normal kidney tissues.The expression and distribution of M1/M2 macrophages in kidney tissues were detected by routine and immunofluorescent staining; mRNA of CD68, CD206 and iNOS detected by polymerase chain reaction(PCR); Murine vascular pericytes subjected to TGF-β1 in vitro and the expressions of α-SMA and PDGFR-β in perivascular cells detected by immunoblotting and cellular fluorescence; The co-culturing models of vascular pericytes and M1/M2 macrophages were constructed.The expressions of α-SMA and PDGFR-β in pericytes were detected by immunoblotting, cellular fluorescence and PCR.Results:A marked infiltration of CD68+ iNOS+ M1 macrophages was present in allograft tissues of recipients with CGD while no obvious infiltration of CD68 + CD206 + was observed.The mRNA levels of CD68, iNOS and CD206 were significantly higher in CGD group than those in control group( P<0.05); In CGD allograft tissues, protein expressions of α-SMA and PDGFR-β spiked markedly( P<0.05)while cells with double staining of α-SMA and PDGFR-β were markedly infiltrated in interstitial area of CGD allograft.TGF-β1 could induce a marked elevation of PMT-related markers in a time-dependent manner( P<0.05); Immunoblotting and cellukar fluorescence indicated that M1 macrophages could promote the elevations of α-SMA and PDGFR-β in pericytes in vitro while M2 macrophages showed no effect on pericyte-to-myofibroblast transition in pericytes. Conclusions:M1 macrophage polarization may promote the formation of renal allograft interstitial fibrosis through promoting PMT.

Healthy Beagle dogs were administrated with batroxobin by intravenous infusion at high, medium and low doses. The study of pharmacodynamics and pharmacokinetics was intended to clarify the relevance of them and provided strong evidence for clinical use of batroxobin. The blood samples were collected after injection based on the time schedule and samples were tested by ELISA method to get the concentration of batroxobin. At the same time, changes of prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (Fib) and D-dimmer were tested. The results showed that the concentration of D-D increased significantly after administration compared with that of before administration. The main pharmacokinetic parameters were as follows: t1/2 were (2.27 +/- 0.42) h, (10.65 +/- 2.19) h and (11.01 +/- 3.51) h; C(max) were (11.9 +/- 1.72) ng x mL(-1), (154.53 +/- 12.38) ng x mL(-1) and (172.14 +/- 47.33) ng x mL(-1); AUC(last) were (29.38 +/- 3.69) ng xh x mL(-1), (148.43 +/- 72.85) ng x h x mL(-1) and (599.22 +/- 359.61) ng x h x mL(-1). The elimination of batroxobin was found to be in accord with linear kinetics characteristics. The results of pharmacodynamics showed that D-dimmer level increased significantly after the administration of batroxobin, which was similar with the changes of batroxobin plasma concentration. Simultaneously, Fib concentrations in Beagle dog blood decreased significantly after the iv administration of batroxobin, while recovered to base level after 48 hours. PT, TT and APTT significantly became longer after administration, which returned to normal level after 48 hours. Especially, the D-dimmer levels and the batroxobin concentration in plasma after intravenous infusion of the drug were synchronized in Beagle dogs. Changes between PD/PK results had obvious correlation, and the D-dimmer levels in plasma can be one of the important monitoring indicators of batroxobin in thrombolytic medication.
Animals ; Area Under Curve ; Batroxobin ; administration & dosage ; blood ; pharmacokinetics ; pharmacology ; Dogs ; Enzyme-Linked Immunosorbent Assay ; Fibrin Fibrinogen Degradation Products ; metabolism ; Fibrinogen ; metabolism ; Fibrinolytic Agents ; administration & dosage ; blood ; pharmacokinetics ; pharmacology ; Infusions, Intravenous ; Male ; Partial Thromboplastin Time ; Prothrombin Time ; Thrombin Time