Objective To explore the prophylactic effect of methylprednisolone combined with granisetron on postoperative nausea and vomiting.Methods Two hundred patients scheduled for lumpectomy of breast were randomly divided into four groups with 50 cases each.The patients in group M1 received a pre-anesthesia intravenous doses of methylprednisolone 25 mg,the patients in group M2 were injected methylpredsisolone 25 mg repeatedly four hours later,in group D received a pre-anesthesia doses of dexamethasone 5 mg,in group N normal saline 2 ml.All the four groups of patients received granisetron 3 mg intravenously at the end of surgery.The incidence of nausea and vomiting in the 24 hours were observed.Results The PONV incidences of group M1,M2,D,N were 36%,18%,38% and 58%.Both group M1,M2 and D significantly decreased the total inci-dence of PONV (P <0.05)in the 24 h.The incidence of PONV was significantly lower in group M2, compared with group M1 and group D respectively (P <0.05).Conclusion Methylprednisolone-gran-isetron combination is as equally effective as dexamethasone-granisetron combination for preventing PONV in lumpectomy,but repeated methylprednisolone after 4 h is more effective than dexametha-sone and single-used methylprednisolone.

Objective To analyze the effect of trametes robiniophila on the apoptosis and the expressions of invasion and metastasis related matrix metalloproteinase 2 (MMP-2) and MMP-9 in human gastric carcinoma poorly differentiated cell line MKN-45 and medium differentiated cell line SGC-7901.Methods Human gastric carcinoma cell lines MKN-45 and SGC-7901 incubated with trametes robiniophila at the different concentrations [0 (the negative control group), 5, 10, and 20 mg/ml] for 24 h. When bifluorescently stained with acridine orange-ethidium bromide (AO-EB), morphology was observed by using microscopy. Flow cytometry was used to test the apoptosis ratio after 24, 48, and 72 h. Reverse transcription polymerase chain reaction (RT-PCR) was applied to analyze the expressions of mRNA of MMP-2 and MMP-9 after 24 h, and the absorbance values of the bands after gel electrophoresis were used as their expression levels. Results Under the fluorescence microscope, the cells of the negative control group were green (normal cells), and the membrane was even in size and integrity. The proportion of apoptotic and necrotic cells was increased with the concentration enrichment of trametes robiniophila. The apoptotic cells were yellow, and their cell membrane lost integrity. Apoptotic bodies were found in cancer cells, and cell membrane showed bud projection. The nuclei of the apoptotic cells showed brightly condensed chromatin or fragmented. Chromatin was strongly stained and located in karyotheca. The necrotic cells were dyed red with integrity of size. The apoptotic ratio of MKN-45 cell line induced by trametes robiniophila after 24 h was (6.5 ±0.8)％, (14.6±1.0)％, (18.0±1.1)％, and (23.1±1.2)％, respectively (F= 333.972, P< 0.01) in negative control group and 5, 10, and 20 mg/ml trametes robiniophila group. After 48 h, the apoptotic ratio was (7.3 ±1.2)％, (18.3 ± 1.6)％, (24.5±1.3)％, and (27.2±1.7)％, respectively (F= 528.432, P= 0.001); after 72 h, the apoptotic ratio was (7.5 ±0.9)％, (50.2 ±1.6)％, (58.0 ±1.9)％, and (69.0 ±1.4)％, respectively (F= 3814.238, P< 0.01). After SGC-7901 cell line induced by trametes robiniophila for 24 h, the apoptotic ratio was (12.9 ±1.0)％, (19.4 ± 1.2)％, (22.0±1.7)％, and (23.0±1.9)％, respectively (F= 120.190, P< 0.01) in negative control group and 5, 10, and 20 mg/ml trametes robiniophila group. For 48 h, the apoptotic ratio was (10.2 ±1.3)％, (40.9 ±1.4)％, (51.6 ±1.9)％, and (66.2 ±1.9)％, respectively (F= 1281.342, P< 0.01). For 72 h, the apoptotic ratio was (27.4 ±1.8)％, (49.7 ±1.4)％, (65.1 ±1.4)％, and (69.0 ±2.0)％, respectively (F= 1112.767, P< 0.01). The induction of apoptosis showed time and dose dependence (both P< 0.01). There was a trendency that the apoptotic ratio of SGC-7901 cell line was higher than that of the MKN-45 cell line at the same condition. RT-PCR showed that mRNA relative expression level of MMP-2 was 0.64±0.02, 0.49±0.01, 0.36±0.02, and 0.32±0.01, respectively (F= 274.321, P< 0.01) of negative control group and 5, 10, and 20 mg/ml trametes extract group after the effect of trametes extract on gastric cancer MKN-45 cell line. The relative expression level of MMP-9 in MKN-45 cell line was 0.71±0.01, 0.54±0.02, 0.47±0.02, and 0.39±0.02, respectively (F=203.948, P< 0.01). The expression level of MMP-2 in SGC-7901 cell line was 0.64±0.01, 0.42±0.02, 0.34± 0.20, and 0.29±0.01, respectively (F= 305.877, P< 0.01), while the mRNA expression level of MMP-9 was 0.65 ±0.15, 0.47 ±0.01, 0.44 ±0.01, and 0.39 ±0.02, respectively (F= 265.259, P< 0.01). Compared with the negative control group, the expression levels of MMP-2 and MMP-9 of the two cell lines were both decreased, after incubated with trametes robiniophila at the different concentrations (all P< 0.05), and with the addition of concentration, the expression levels of MMP-2 and MMP-9 were decreased. Conclusion Trametes robiniophila can induce the apoptosis of human gastric carcinoma cell lines MKN-45 and SGC-7901 in vitro and can inhibit the expressions of MMP-2 and MMP-9 and the effect of trametes robiniophila may be related with the differentiation degree.

8-prenylnaringenin (8-PN) is a potent estrogen with high medicinal values. It also serves as an important precursor for many prenylated flavonoids. Microbial synthesis of 8-PN is mainly hindered by the low catalytic activity of prenyltransferases (PTS) and insufficient supply of precursors. In this work, a SfN8DT-1 from Sophora flavescens was used to improve the efficiency of (2S)-naringenin prenylation. The predicted structure of SfN8DT-1 showed that its main body is comprised of 9 α-helices and 8 loops, along with a long side chain formed by nearly 120 amino acids. SfN8DT-1 mutants with different side-chain truncated were tested in Saccharomyces cerevisiae. A mutant expressing the truncated enzyme at K62 site, designated as SfND8T-1-t62, produced the highest 8-PN titer. Molecular docking of SfN8DT-1-t62 with (2S)-naringenin and dimethylallyl diphosphate (DMAPP) showed that K185 was a potentially crucial residue. Alanine scanning within a range of 0.5 nm around these two substrates showed that the mutant K185A may decrease its affinity to substrates, which also indicated K185 was a potentially critical residue. Besides, the mutant K185W enhanced the affinity to ligands implied by the simulated saturation mutation, while the saturated mutation of K185 showed a great decrease in 8-PN production, indicating K185 is vital for the activity of SfN8DT-1. Subsequently, overexpressing the key genes of Mevalonate (MVA) pathway further improved the titer of 8-PN to 31.31 mg/L, which indicated that DMAPP supply is also a limiting factor for 8-PN synthesis. Finally, 44.92 mg/L of 8-PN was produced in a 5 L bioreactor after 120 h, which is the highest 8-PN titer reported to date.
Dimethylallyltranstransferase/metabolism* ; Flavonoids/metabolism* ; Molecular Docking Simulation ; Prenylation ; Saccharomyces cerevisiae/metabolism* ; Sophora/metabolism*

ObjectiveTo explore the effect of oleanolic acid （OA） on water metabolism in mice with water-dampness retention caused by spleen deficiency and the mechanism. MethodThe 60 SPF Kunming （KM） mice were randomized into blank group （n=10） and modeling group （n=50）. Through long-term living in damp place and irregular diet， water-dampness retention caused by spleen deficiency was induced in modeling mice. Then the model mice were randomly classified into model group， natural recovery group， and low-dose， medium-dose， and high-dose OA groups. The mice in the blank group， model group， and natural recovery group were given （ig） 10 mL·kg^-1·d^-1 normal saline， and mice in the low-dose， medium-dose， and high-dose OA groups received 50， 100， 200 mg·kg^-1·d^-1 OA， respectively. The intervention lasted 7 days. Before and after modeling and administration， the general conditions of the mice were observed and body weight of mice was measured. The water content in feces and tissues was detected with the oven-drying method， and water load index and organ coefficient were measured with the weighing method. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the urinary D-xylose excretion， serum gastrin （GAS）， total protein （TP）， albumin （ALB）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， interleukin-6 （IL-6）， antidiuretic hormone （AVP）， aquaporin 1 （AQP1） in renal medulla， and liver Na⁺-K⁺-ATPase. At the same time， OA was docked with ALB， IL-6， AQP1， and Na⁺-K⁺-ATPase. ResultCompared with the blank group， the model group showed withered hair， emaciation， laziness， bradykinesia， slow weight growth， infrequent spontaneous activities， high water content in feces and tissues， low weight loss after water loading， high coefficient of each organ （P<0.05， P<0.01）. Moreover， the model group had less urinary D-xylose excretion， lower serum levels of GAS， TP， ALB， and HDL-C， higher levels of TC， LDL-C， AVP， and IL-6， lower expression of Na⁺-K⁺-ATPase in the liver， and higher expression of AQP1 in renal medulla than the blank group （P<0.05， P<0.01）. The three OA groups demonstrated better general conditions， faster weight gain， more frequent spontaneous activities， lower water content in feces and tissues， larger weight loss after water loading， and lower coefficient of each organ than the model group （P<0.05， P<0.01）. Moreover， compared with the model group， the three OA groups had high D-xylose excretion， high serum levels of GAS， TP， ALB， and HDL-C， low serum levels of TC， LDL-C， AVP， and IL-6， high expression of Na⁺-K⁺-ATPase in liver， and low expression of AQP1 in renal medulla （P<0.05， P<0.01）. The recovery in each OA group was better than that in natural recovery group. Molecular docking results also confirmed that OA had high binding affinity with ALB， IL-6， AQP1， and Na⁺-K⁺-ATPase. ConclusionOA can alleviate the abnormal water metabolism in mice with water-dampness retention caused by spleen deficiency， which lays a basis for its potential clinical application.