Objective To investigate the effects of ketamine on the voltage-gated sodium channels in hippocampal pyramidal neurons, trying to elucidate the possible mechanism of general anesthesia with ketamine.Methods Hippocampal pyramidal neurons were isolated from Wistar rats of 2 weeks old. Whole-cell patch-clamp recordings were made from cultured hippocampal pyramidal neurons before and after the application of ketamine The effect of ketamine on the sodium current amplitude and the kinetics of the channel were studied. Results The sodium channels were reversibly inhibited by ketamine in a dose-dependent manner. IC50 of ketamine was (794?21) ?mol/L.The hyperpolarizing shift of both steady-activation and steady-inactivation was observed.Conclusion Ketamine inhibits the voltage-gated sodium channels to some degree. Sodium channel inhibition may be involved in the mechanism of general anesthesia induced with ketamine.

Ergosterol is an economically important metabolite produced by yeast. To improve the production of ergosterol by Saccharomyces cerevisiae YEH56 (pHXA42) from molasses, a cheap and regenerative material, different strategies were applied. First, Plackett-Burman design and central composite design were applied to screen the significant factors in fermentation medium using ergosterol yield (g/L) as the response value. Ergosterol yield reached 371.56 mg/L by using the optimal fermentation medium in shake-flask culture (total sugar in molasses 40 g/L, KH2PO4 1 g/L, K2HPO4 1.86 g/L, CuSO4 x 5H2O 17.5 mg/L, FeSO4 x 7H2O 13.9 mg/L, MgSO4 x 5H2O 12.3 mg/L, corn steep liquor 10 mL/L), which was increased by 29.5% compared with the initial culture. Second, ergosterol yield was increased by 62.1% using a pH-control strategy in a 5-L bioreactor. Third, ergosterol production was improved further by using molasses feeding strategy. After 38 h fermentation, ergosterol yield reached 1 953.85 mg/L, which was 3.2 times of that in batch fermentation. Meanwhile, ergosterol production rate was increased by 42.7% compared with that in the batch culture.
Culture Media ; Ergosterol ; biosynthesis ; Fermentation ; Industrial Microbiology ; Molasses ; Saccharomyces cerevisiae ; genetics ; growth & development ; metabolism

Objective To investigate the effect of targeted monitoring on the control of central venous catheter associated infection, study the difference of CVC?RI infection rate and the distribution of pathogenic bacteria in the general hospital and the maternal and child hospital, and analyse the main pathogenic bacteria drug resistance characteristics. Methods From January 2013 to December 2014,685 cases patients with central venous catheterization in the Port Hospital of Hebei Port Group Company Limited,and four cooperative hospitals ( Affiliated Hospital of North China University of Science and Technology, People′s Hospital of Tangshan, Tangshan Port Economic Development District Hospital, Maternal and Child Health Care Hospital of Laoting County) were selected as the research objects. The difference of infection rate before and after the targeted monitoring in 2014 was compared. The tip of the catheter and the peripheral blood were cultured in the patients with suspected infection,bacterial identification and drug sensitivity test were carried out using Compact VITEK2 automatic bacteria identification/drug sensitivity analyzer. Results The infection rate of CVC?RI of five hospitals was 18. 75%( 60/320 ) in 2013, 11. 51%( 42/365 ) in 2014, the difference of infection rate was statistically significant before and after the implementation of the target monitoring(χ2=7. 059,P<0. 05) . The CVC?RI infection rate was 15. 52%(81/522) in the general hospital,12. 88%(21/163) in the maternal and child hospital,the difference was not statistically significant(P>0. 05). In 102 cases of CVC?RI,105 bacterial strains were detected in 91 cases,gram positive bacteria accounted for 51. 43%( 54/105) ,gram negative bacilli accounted for 40. 95%( 43/105) ,fungi accounted for 7. 62%( 8/105) . There was no significant difference in the proportion of pathogenic bacteria in the general hospital and the maternal and child hospital(P>0. 05). Grampositive cocci were mainly staphylococcus aureus and coagulase negative staphylococci,they were completelysensitive to linezolid, quinupristin/ dalfopristin, tigecycline, and vancomycin. They were high sensitivity torifampicin,while highly resistant to gentamicin, tetracycline, penicillin; Gram negative bacilli were mainlypseudomonas aeruginosa and escherichia coli,they were high sensitivity to meropenem,imipenem,while resistantto multiple antimicrobia1s. Conclusion Adhere to the implementation of targeted monitoring help to reduce theoccurrence of CVC?RI. The isolation of pathogenic bacteria from multiple drug resistance should be paid moreattention.

Objective To explore the clinical characteristics and treatment in elderly patients with gastric bezoars. Methods Totally 49 patients with gastric bezoars were retrospectively analyzed,and among them,36 patients were aged 65-77 years, 48 cases underwent sequential therapy,1 elderly patient underwent surgical operation. Results There were differences in etiology, medical histories, clinical manifestations and therapeutic efficacy between elderly and non elderly patients with gastric bezoars.There were 42％ (15/36) of elderly patients eating glutinous rice cakes,4 cases with gastric surgery histories,6 cases with diabetes in elderly patients,but in non eldcrly patients only 8％ (1/13) of patients eating glutinous rice cake,and there were no cases with surgery histories and diabetes. Gastric bezoars in 48 cases were cleared away by sequential therapy. 1 elderly patient developed intestinal obstruction after surgical operation and died of infectious shock. Conclusions There are some different characteristics between elderly and nonelderly patients with gastric bezoars.Sequential therapy is safe and effective for elderly patients with gastric bezoars.

Objective To investigate the expression of SOX4 and C/EBPα mRNA in chronic myeloid leukemia (CML) and their clinical significances. Methods Bone marrow samples from 68 cases of CML including 57 newly diagnosed patients and 11 patients treated with imatinib were collected, and peripheral blood mononuclear cells from 30 healthy people were collected as healthy control. The expression of SOX4 and C/EBPαmRNA and protein levels were detected by RT-PCR and Western blot, respectively. The relations between the expression of SOX4 and C/EBPα and the influences of imatinib on SOX4 and C/EBPα were analyzed. Results The expression level of SOX4 mRNA was increased in newly diagnosed CML patients compared with that of normal control group (6.545 5±1.495 2 vs. 0.059 6±0.018 8, t=3.139, P=0.002 3), but the expression level of C/EBPαmRNA was significantly decreased (0.238 8±0.033 8 vs. 0.810 5±0.056 2, t=9.240, P<0.000 1). The expression levels of SOX4 and C/EBPαmRNA had no significant correlation with age, gender, white blood cell count (WBC) and bcr-abl of newly diagnosed patients (all P>0.05). The expression level of SOX4 mRNA in 5 patients treated with imatinib was decreased (0.120 6 ±0.044 9 vs. 0.557 9±0.144 8, t=2.885, P=0.020 4), and the expression level of C/EBPαmRNA was increased (0.330 3±0.042 4 vs. 0.150 5±0.046 5, t=2.855, P=0.021 3). The expression level of SOX4 mRNA in 6 patients who developed blast phase during the treatment of imatinib was increased (0.469 9±0.123 0 vs. 0.050 2±0.036 6, t=2.370, P=0.039 3), and the expression level of C/EBPα mRNA was decreased (0.197 9 ±0.064 7 vs. 0.378 7±0.042 9, t=2.327, P=0.042 3). The expression of SOX4 mRNA was negatively correlated with C/EBPα mRNA (t=-0.554 6, P=0.002 8). Conclusions In newly diagnosed CML, the expression level of SOX4 is increased, C/EBPα is decreased compared with that of healthy control, and both have negative correlation. In the patients in blast phase after imatinib treatment, SOX4 gene is up-regulated, and C/EBPα is down-regulated. C/EBPα-SOX4 axis may play a role in the occurrence and development of CML. SOX4 may be a new molecular target for the treatment of CML.

Objective:To summarize and compare the characteristics of oral microbiota in women during the preconception period and the third trimester.Methods:This retrospective cohort study involved 55 women who were recruited in the Preconceptional Offspring Trajectory Study (PLOTS) conducted by Fudan University and followed up to the third trimester in the Maternal and Child Health Care Hospital of Jiading District of Shanghai from September 2016 to December 2019. A total of 110 unstimulated saliva samples were collected in the preconception period ( n=55) and the third trimester ( n=55). Features of oral microbiota in the samples were analyzed by 16S rRNA gene-based sequencing. Moreover, the related factors were also analyzed. Paired t test or Wilcoxon matched-pairs signed-ranks test were used to analyze the differences in α-diversity during preconception and the third trimester; t test, analysis of variance (ANOVA), Kruskal-Wallis test and Mann-Whitney U test for comparison between groups with different characteristics and permutational multivariate analysis of variance (PerMANOVA) for β-diversity were used; Linear discriminant analysis (LDA) effect size (LEfSe 1.0) was used to identify the iconic oral flora. Results:(1) The Ace index of oral microbiota was significantly lower in the third trimester than that in the preconception period [661.14(578.15-752.85) vs 730.64 (632.40-911.00), T=1 077.00, P=0.010]. There was also a significance difference in β-diversity ( F=12.539, R2=0.104, P=0.001). Some species such as Saccharibacteria_TM7_G3, Prevotella_7, Absconditabacteria_SR1_G1, Porphyromonas, Ruminococcaceae_UCG_014, Prevotella, Peptostreptococcus, Prevotella_2, Alloprevotella, Parvimonas, Solobacterium and Eubacterium_nodatum_group in saliva were statistically more abundant in the third trimester than those in the preconception period (all P<0.05). (2) The third-trimester Shannon index was lower among those with lower income [5.44 (5.08-5.77) vs 5.75 (5.44-6.12), U=219.00, P=0.029] and those with gargle habit after meal or dessert [5.36 (4.91-5.48) vs 5.72 (5.44-6.05), U=374.00, P=0.046]. Conclusions:The features of oral microbiota vary in women during the preconception period and the third trimester. There is a significant increase in the abundance of oral pathogenic and opportunistic bacteria in the third trimester.

The heterologously expressed L1 protein of human papilomavirus 16 can assembly into virus-like particles (VLPs), which has been used as prophylactic vaccine for cervical carcinoma. To express L1 protein in Hansenula polymorpha, we analyzed the codon usage of the native gene of L1 protein and redesigned the encoding sequence according to the codon bias of H. polymorpha. We used assembly PCR to synthesize the native gene HPV16L1-N and the codon optimized gene HPV16L1. The synthesized genes were cloned into pMOXZa-A vector to generate plasmids pMOXZ-HPV16N and pMOXZ-HPV16. The expression cassettes MOXp-HPV16L1(N)-AOXTT were cloned into YEp352 vector and transferred into H. polymorpha. After methanol inducement, the expression of L1 protein in H. polymorpha was detected from the codon optimized gene HPV16L1 rather than the native gene HPVI6L1-N. The parameters for induced cultivation for strain HP-U-16L with HPV16L1 were investigated in shaking flask cultures. After induced cultivation in YPM (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 12 h at 37 degrees C for 72 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by H. polymorpha.
Capsid Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Codon ; genetics ; Genetic Vectors ; genetics ; Human papillomavirus 16 ; genetics ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

Yeast flocculation is described as a reversible, asexual and calcium dependent process, in which cells adhere to form flocs by interaction of specific cell surface proteins named flocculins on yeast cells with mannose residues present on the cell wall of adjacent yeast cells. Yeast flocculation provides a very economical and convenient pathway for separation of yeast cells from the fermentation broth or removal of heavy metal ions from effluent. A large number of tandem repeats have been found in genes encoding flocculins, which not only have great regulatory effect on the structure and function of flocculins, generating the diversity of flocculation characteristics, but lead to genetic instability in flocculation as well for driving slippage and recombination reactions within and between FLO genes. Here, the research progress in effect of variation of tandem repeats in FLO genes on flocculation characteristics and genetic stability were reviewed to direct and promote the controllable application of flocculation in industrial fermentation process and environmental remediation.
Fermentation ; Flocculation ; Mannose ; Membrane Proteins ; genetics ; Saccharomyces cerevisiae ; genetics ; growth & development ; Saccharomyces cerevisiae Proteins ; genetics ; Tandem Repeat Sequences

Objective:To investigate the value of ultrasound elastography combined with serum urokinase plasminogen activator (uPA) in differential diagnosis of benign and malignant thyroid nodules.Methods:From May 2018 to May 2019, 150 patients with thyroid nodules (160 nodules) were selected, 69 patients with benign nodules (76 benign nodules) and 81 patients with malignant nodules (84 malignant nodules) were diagnosed by postoperative pathology. In the same period, 40 healthy people underwent health examination were selected as the control group. All preoperative patients were subjected to ultrasound elastography. The results of pathological examination were diagnosed as "gold standard"; the characteristics of ultrasound elastography were observed and scored, and compared with the results of pathological examination. The level of serum uPA was measured by enzyme-linked immunosorbent assay (ELISA), the sensitivity, specificity, and the area under the receiver operating characteristic (ROC) curve (AUC) were compared among single real-time ultrasound elastography, serum uPA and combined diagnosis.Results:Among the 150 patients (160 nodules), for ultrasound elastography, the sensitivity was 97.37% (74/76), the specificity was 85.71% (72/84), the accuracy was 91.25% (146/160), and the Kappa value was 0.726. The ratio of 0 to 2 points in malignant group was significantly lower than that in the benign group, the ratio of 3 to 4 points was significantly higher than that in the benign group ( P < 0.05). Compared with the control group, the levels of serum uPA in the benign group and the malignant group increased significantly ( P < 0.05). Compared with the benign group, serum uPA level in the malignant group increased significantly ( P < 0.05). The AUC of combined diagnosis was 0.869, 95% confidence interval ( CI): 0.789 - 0.949, and the best cut-off point for the combined diagnosis was uPA = 982.16 mU/L, at this time, the diagnostic sensitivity was 0.646, and the diagnostic specificity was 0.575. The AUC of ultrasound elastography was 0.814, 95% CI: 0.721 - 0.907, and the AUC of combined diagnosis was higher than that of ultrasound elastography. Conclusion:The clinical value of ultrasound elastography combined with serum uPA in differentiating benign and malignant thyroid nodules is higher than that of ultrasound elastography.

Objective:To analyze the diagnosis of renal injury caused by chronic arsenism by color Doppler ultrasound and the observation of renal hemodynamics.Methods:From January 2018 to April 2019, 74 patients with occupational chronic arsenism admitted to Binzhou People's Hospital were selected and divided into two groups according to whether the patients were complicated with renal injury or not, 26 patients in renal injury group and 48 patients in non-renal injury group. The renal function indexes [serum creatinine (SCr), blood urea nitrogen (BUN), urine and blood β2-microglobulin (β2-MG)] were measured and the renal hemodynamics [peak blood flow velocity during systole (Vmax), minimum blood flow velocity during diastole (Vmin), resistance index (RI)] of the two groups were analyzed by color Doppler ultrasound, and the correlation between renal function indexes and renal hemodynamics was analyzed.Results:The levels of SCr, BUN, urine β2-MG and blood β2-MG in renal injury group were significantly higher than those in non-renal injury group [(136.28 ± 21.05) vs (108.42 ± 26.49) μmol/L, (8.03 ± 1.04) vs (5.36 ± 0.97) mmol/L, (157.48 ± 25.63) vs (127.42 ± 18.95) μg/L, (3.97 ± 1.12) vs (2.35 ± 0.84) mg/L, t = 4.625, 11.022, 5.740, 7.028, P < 0.05]. The Vmax and Vmin of the main renal artery and interlobular artery in renal injury group were significantly lower than those in non-renal injury group [cm/s: (50.34 ± 13.42) vs (75.32 ± 16.52), (18.13 ± 5.21) vs (29.83 ± 7.05), (12.31 ± 3.82) vs (17.22 ± 5.07), (4.08 ± 1.15) vs (6.03 ± 1.93), t = 6.833, 7.084, 4.657, 4.775, P < 0.05], and the RI of the main renal artery and interlobular artery in renal injury group was significantly higher than that in non-renal injury group [(0.71 ± 0.06) vs (0.62 ± 0.08), (0.68 ± 0.10) vs (0.56 ± 0.08), t = 5.017, 5.634, P <0.05]. Vmax and Vmin of the main renal artery and interlobular artery were negatively correlated with SCr, BUN, urine β2-MG and blood β2-MG ( P < 0.05). RI of the main renal artery and interlobular artery were positively correlated with SCr, BUN, urine β2-MG and blood β2-MG ( P < 0.05). Conclusions:Color Doppler ultrasound can effectively reflect the status of renal injury in chronic arsenism patients. The renal blood flow perfusion in renal injury patients is decreased, and the blood flow resistance is increased, which is closely related to the degree of renal injury and deserves clinical attention.