Objective: To clarify the specific mechanisms of action of raw Phellodendron chinense Schneid. (RPC) and saltwater-processed PC (SPC) in the treatment of rats with a kidney-yin deficiency pattern (KYDP). Methods: Healthy rats were administered hydrocortisone to establish a KYDP model. The rats were divided into seven groups: blank control, model, positive control (Liuwei Dihuang pills), high-dose RPC, low-dose RPC, high-dose SPC, and low-dose SPC. Enzyme-linked immunosorbent assay was used to measure the levels of cAMP, cGMP, TRH, TSH, T3, T4, IFN-γ, TNF-α, and testosterone in the serum and the levels of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in the liver. TRH mRNA expression in the rat hypothalamus was measured using RT-PCR. THRα1+2 protein expression in the hypothalamus of rats was measured using Western blot. Immunohistochemistry was performed to determine the expression levels of FAS, FasL, and TSHR. Flow cytometry was used to determine CD4+ and CD8+ T lymphocyte levels. Illumina MiSeq sequencing technology was used to evaluate the diversity of intestinal flora in KYDP rats. Results: The cAMP/cGMP ratio was significantly higher in the model group than in the blank control group (P = .048). Compared with the model group, after administration, the levels of the above-mentioned serum and liver indexes decreased, except that of testosterone. The CD4+/CD8+ ratio also decreased. Compared with the RPC group, the levels of T3, IFN-γ, FAS, FasL, and TSHR in the SPC group decreased whereas that of testosterone increased. Additionally, immune function and intestinal flora diversity improved in the SPC group. SPC proved to be more effective in improving liver energy metabolism in KYDP rats than RPC. Conclusion SPC had a better therapeutic effect on KYDP than RPC. The underlying mechanism of action may be related to improvements in liver energy metabolism, immune function, and intestinal flora diversity.

Gluconacetobacter xylinus is a primary strain producing bacterial cellulose (BC). In G. xylinus, BcsD is a subunit of cellulose synthase and is participated in the assembly process of BC. A series of G. xylinus with different expression levels of the bcsD gene were obtained by using the CRISPR/dCas9 technique. Analysis of the structural characteristics of BC showed that the crystallinity and porosity of BC changed with the expression of bcsD. The porosity varied from 59.95%-84.05%, and the crystallinity varied from 74.26%-93.75%, while the yield of BC did not decrease significantly upon changing the expression levels of bcsD. The results showed that the porosity of bacterial cellulose significantly increased, while the crystallinity was positively correlated with the expression of bcsD, when the expression level of bcsD was below 55.34%. By altering the expression level of the bcsD gene, obtaining BC with different structures but stable yield through a one-step fermentation of G. xylinus was achieved.
Cellulose/chemistry* ; Clustered Regularly Interspaced Short Palindromic Repeats ; Fermentation ; Gluconacetobacter xylinus/metabolism*

Objective:To investigate the prognostic value of infarct size (IS) in patients with acute ST-segment elevation myocardial infarction (STEMI) underwent primary percutaneous coronary intervention (PCI).Methods:A total of 104 patients with acute STEMI who underwent primary PCI treatment in Shengjing Hospital of China Medical University from February 2017 to November 2018 were included in the present study. All patients underwent cardiovascular magnetic resonance (CMR) within one week after primary PCI treatment. The subjects were followed up for two years. Major adverse cardiac events (MACE) included new onset congestive heart failure and/or recurrent nonfatal myocardial infarction and/orcardiac death. The optimal IS cutoff value for MACE was determined by receiver operating character (ROC) curve. Based on the IS cutoff value, the patients were divided into the high IS group and the low IS group. Clinical characteristics between the two groups were compared. A cox regression model was used to analyze the prognostic value of IS in acute STEMI patients treated with primary PCI for the adverse events.Results:The IS cutoff value determined by ROC curve was 13.55%. 50 patients were in the high IS group (IS≥13.55%) and 54 patients were in the low IS group (IS<13.55%). More female patients [14 cases (28.0%) vs. 6 cases (11.1%)] were in the IS group, and a higher proportion of patients in the high IS group had anterior myocardial infarction [27 cases (54.0%) vs. 16 cases (29.6%)] or microvascular obstruction [32 cases (64.0%) vs. 16 cases (29.6%)]. White blood cell counts [11.25(8.90, 13.38) ×10 9/L vs. 9.25(7.58, 11.00) ×10 9/L], troponin I levels [50.63(16.56, 76.30)μg/L vs. 16.58(2.66, 38.42)μg/L] and brain natriuretic peptide levels [178.10(79.70, 281.95)μg/L vs. 79.60(42.83, 183.90)μg/L] in the high IS group were higher than those in the low IS group ( P<0.05), and left ventricular ejection fraction [(45.15±10.65)% vs. (51.95±12.91)%] in the high IS group was lower than that in the low IS group ( P<0.05). Multivariate Cox regression analyses showed that IS was independently associated with the risk of cardiac death in patients with acute STEMI two years after primary PCI( P=0.033, HR=1.075, 95% CI1.006-1.148). Every 1% increase in IS was associated with a 7.5% increase in cardiac death. Conclusions:Infarct size, measured by CMR within one week after primary PCI, is strongly associated with cardiac death in patients with acute STEMI two years after primary PCI. IS could be used as an index for the prognosis of patients with acute STEMI.

OBJECTIVE：To study the prepar ation technology of gastric floating tablets of Schisandra chinensis total lignans （SCTL），and evaluate the quality of prepared tablets. METHODS ：Based on single factor test ，the orthogonal experiment was conducted to optimize the formulation of SCTL gastric floating tablets with the contents of hydroxypropylmethylcellulose （HPMC） K15M，NaHCO3 and microcrystalline cellulose as the factors ，using starting time ，holding time and cumulative release rate of gastric floating tablets as evaluation indexes. The properties ，weight difference ，floatability and accumulative release rate of the prepared SCTL gastric floating tablets were determined. The gastric floating tablets were qualitatively identified by TLC ，and the contents of schisandrin A and total lignans were determined by HPLC and UV spectrophotometry. RESULTS ：The optimal formulation of SCTL gastric floating tablets was made up of 23％ SCTL extract ，20％ HPMC K 15M，40％ microcrystalline cellulose，15％ sodium bicarbonate ，1％ octadecyl alcohol and 1％ polyvinylpyrrolidone. The results of detection of this preparation were in line with the related provisions of “0101 tablet”stated in 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. TLC indicated that the chromatogram of the test sample showed the main spots of same color as the corresponding positions of the chromatogram of schizandrol A control ，Schisandra chinensis reference substance and raw material ，while the negative control has no interference. Content determination results shows that the average content of schizandrol A and total lignans in SCTL gastric floating tablets is 3.187，19.617 mg. It was preliminarily formulated that the content limitation of schizandrol A in one tablet should not be less than 2.50 mg，and the content of total lignans （calculated by schizandrol A ）should not be less than 15.50 mg. CONCLUSIONS：The preparation technology of SCTL gastric floating tablets is stable ，feasible and controllable in quality.

Objective To study the effect of deep inspiration breath-hold(DIBH)technique on the heart dose in whole breast irradiation(WBI)for left breast cancer after breast-conserving surgery, and to investigate the anatomical factors for heart dose. Methods Fifteen patients with left breast cancer who received WBI after breast-conserving surgery and met breathing control requirements were prospectively enrolled as subjects. Simulated CT scans were performed during free breathing(FB)and DIBH. The WBI plans were optimized based on DIBH images.The position,volume,and radiation doses to the heart and lung were compared between the status of FB and DIBH. Correlation of heart dose with various anatomical factors was analyzed in FB status. Between-group comparison of categorical data was made by nonparametric Wilcoxon rank test.A two-variable correlation analysis was made by the Pearson method.Results There was no significant difference in heart volume between the status of FB and DIBH(P=0.773).The volume of both lungs was significantly larger in DIBH status than in FB status(P=0.001). The mean and maximum doses and V5-V40for the heart,left anterior descending coronary artery,left ventricle,right ventricle,and left lung were significantly lower in DIBH status than in FB status(all P<0.05). The greater DIBH increased the lung volume,the greater the mean heart dose decreased. In FB status,the left breast volume,heart-to-lung volume ratio,distance between the inferior margins of breast and heart,and maximum heart margin distance showed a linear correlation with heart dose. Particularly, the heart-to-lung volume ratio and maximum heart margin distance were independently correlated with heart dose. Conclusions DIBH technique in WBI for left breast cancer after breast-conserving surgery significantly reduces heart and lung doses compared with FB. Changes in lung volume are the basis for improving the relative anatomical location of the heart. The heart-to-lung volume ratio and maximum heart margin distance may provide a reference for DIBH technique.

Objective To investigate the quantity and function of CD8+T cells in peripheral blood of pa-tients with repeated implantation failure(RIF).Methods Thirty-seven patients with RIF and 19 healthy controls were enrolled in this study.The peripheral blood and endometrium were collected during the mid-luteal phase.The percentage of peripheral CD8+T subsets and the levels of perforin and granzyme B of peripheral CD8+T cells were determined by flow cytometry assay.The percentage of endometrial CD8+T cells was detected by IHC,the produc-tion of perforin and granzyme B of endometrial CD8+T cells was detected by IF. Results Compared with the con-trol group,the percentage of peripheral CD8+T cells in patients with RIF was not significantly changed(37.22% vs. 37.15%,P>0.05).However,the porportion of endometrial CD8+T cells in the RIF group was higher than that in the control group(1.99% vs.3.77%,P<0.001).The levels of perforin and granzyme B in peripheral blood and en-dometrial CD8+T cells in patients with RIF were similar with those in the control group.Conclusions Compared to the control group,the percentage of endometrial CD8+T was markedly upregulated in patients with RIF.However, the production of perforin and granzyme B were similar between the control group and the RIF group.

Gluconobacter oxydans is known to oxidize glucose to gluconic acid (GA), and subsequently, to 2-keto-gluconic acid (2KGA) and 5-keto-gluconic acid (5KGA), while 5KGA can be converted to L-(+)-tartaric acid. In order to increase the production of 5KGA, Gluconobacter oxydans HGI-1 that converts GA to 5KGA exclusively was chosen in this study, and effects of carbon sources (lactose, maltose, sucrose, amylum and glucose) and nitrogen sources (yeast extract, fish meal, corn steep liquor, soybean meal and cotton-seed meal) on 5KGA production were investigated. Results of experiment in 500 mL shake-flask show that the highest yield of 5KGA (98.20 g/L) was obtained using 100 g/L glucose as carbon source. 5KGA reached 100.20 g/L, 109.10 g/L, 99.83 g/L with yeast extract, fish meal and corn steep liquor as nitrogen source respectively, among which the optimal nitrogen source was fish meal. The yield of 5KGA by corn steep liquor is slightly lower than that by yeast extract. For the economic reason, corn steep liquor was selected as nitrogen source and scaled up to 5 L stirred-tank fermentor, and the final concentration of 5KGA reached 93.80 g/L, with its maximum volumetric productivity of 3.48 g/(L x h) and average volumetric productivity of 1.56 g/(L x h). The result obtained in this study showed that carbon and nitrogen sourses for large-scale production of 5KGA by Gluconobacter oxydans HGI-1 were glucose and corn steep liquor, respectively, and the available glucose almost completely (85.93%) into 5KGA.
Bioreactors ; Carbon ; chemistry ; Culture Media ; chemistry ; Fermentation ; Gluconates ; metabolism ; Gluconobacter oxydans ; metabolism ; Industrial Microbiology ; Nitrogen ; chemistry

Asymmetric reduction of bulky diaryl ketones is still one of the challenging tasks in biocatalysis. By genomic data mining, a putative carbonyl reductase gene pascr was found in Pichia pastoris GS115. pascr was cloned and over-expressed in Escherichia coli Rosseta2 (DE3). The recombinant enzyme was purified to homogeneity by Ni-NTA column and its catalytic properties were studied. PasCR strictly used NADPH as cofactor, gel filtration and SDS-PAGE analysis suggested that the native form of PasCR was a dimmer. PasCR exhibited the highest activity at 35 degrees C in phosphate buffer at pH 6.5. The enzyme catalyzed the reduction of some bulky diaryl ketones, such as 4-methylbenzophenone, 2-methylbenzophenone and 4-chlorobenzophenone, especially for 4-methylbenzophenone, the product S--alcohol was obtained with 85% ee.
Alcohol Oxidoreductases ; genetics ; Amino Acid Sequence ; Catalysis ; Cloning, Molecular ; Ketones ; chemistry ; Molecular Sequence Data ; Pichia ; enzymology ; genetics ; Stereoisomerism

OBJECTIVETo investigate the role of N-Methyl-D-aspartic acid (NMDA)-type glutamate receptors in the central nucleus of the amygdale (CeA) in food and water intake.

METHODSMale Sprague-Dawley rats with stainless steel cannulae implanted unilaterally into the CeA were used. The prototypic NMDA receptor agonist NMDA, or the selective NMDA receptor antagonist D(-)-2-amino-5-phosphonopentanoic acid (D-AP-5) was microinjected into the CeA of satiated and euhydrated rats.

RESULTSIntra-CeA injection of 8.50, 17.00, or 34.00 nmol NMDA did not alter food intake but significantly increased water intake 0-1 h after the injection (F(3,32)=3.191, P=0.037) independent of food intake. Without affecting the food intake, injection of 6.34, 12.70, or 25.40 nmol D-AP-5 into the CeA significantly decreased water intake 0-1 h after the injection (F(3,28)=3.118, P=0.042) independent of food intake.

CONCLUSIONNMDA receptors in the CeA may participate in the control of water intake rather than food intake.

2-Amino-5-phosphonovalerate ; pharmacology ; Amygdala ; drug effects ; Animals ; Drinking ; drug effects ; Eating ; drug effects ; Excitatory Amino Acid Agonists ; pharmacology ; Excitatory Amino Acid Antagonists ; pharmacology ; Injections, Intraventricular ; Male ; N-Methylaspartate ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; agonists ; antagonists & inhibitors

OBJECTIVETo explore the role of µ-opioid receptors (MOR) in the central nucleus of the amygdala (CeA) in modulating sucrose solution intake in rats.

METHODSSprague-Dawley rats received intra-CeA injection of MOR agonist DAMGO or saline, and then underwent two bottle choice test between sucrose solution and distilled water. After intake of sucrose solution or distilled water, activated neurons in the CeA were labeled and identified with MOR/Fos-double labeling immunohistochemistry.

RESULTSCompared with saline injection, intra-CeA injection of DAMGO significantly increased sucrose solution intake in rats over a 3-h period. Sucrose solution intake induced significantly more c-Fos and MOR/Fos double-labeled neurons in the CeA than distilled water intake.

CONCLUSIONSThe CeA participates in modulation of sucrose intake in rats, and MOR may partly mediate this mechanism.

Amygdala ; metabolism ; Animals ; Male ; Neurons ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, mu ; metabolism ; Sucrose ; metabolism