A preliminary study on salivary microbiota of patients with laryngopharyngeal reflux / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the characteristics of salivary microbiota in patients with laryngopharyngeal reflux (LPR). Methods: A case-control study was applied to enroll 60 patients and healthy subjects who were outpatients of the Department of Otorhinolaryngology Head and Neck Surgery of the Eighth Medical Center of the PLA General Hospital from December 2020 to March 2021, including 35 males and 25 females, aged from 21 to 80 (33.75±11.10) years. Thirty patients with suspected laryngopharyngeal reflux were selected as study group and thirty healthy volunteers without pharyngeal symptoms were selected as control group. Their salivary samples were collected, and the salivary microbiota was detected and analyzed by 16S rDNA sequencing. SPSS 18.0 software was used for statistical analysis. Results: There was no significant difference in the diversity of salivary microbiota between the two groups. At the phylum classification level, the relative abundance of Bacteroidetes in the study group was higher than that in the control group[37.86(31.15, 41.54)% vs 30.24(25.51, 34.18)%,Z=-3.46,P<0.01]. And the relative abundance of Proteobacteria in the study group was lower than that in the control group [15.76(11.81, 20.17)% vs 20.63(13.98, 28.82)%, Z=-1.98,P<0.05]. At the genus level, the relative abundance of Prevotella, Lactobacillus, Parascardovia and Sphingobium in the study group was higher than that in the control group(Z values were-2.92, -2.69, -2.05, -2.31, respectively, P<0.05).And the relative abundance of Streptococcus, Cardiobacterium, Klebsiella and Uruburuella of study group was lower than that of control group(Z values were -2.43, -2.32, -2.17, -2.32, respectively, P<0.05). LEfSe difference analysis showed that there were 39 bacteria with significant differences between the two groups, including Bacteroidetes, Prevotellaceae and Prevotella, which were enriched in the study group, and Streptococcaceae, Streptococcus and other taxa, which were enriched in the control group(P<0.05). Conclusion: The changes of the microflora in the saliva between LPR patients and healthy people suggest that the dysbacteriosis might exist in LPR patients, which may play an important role in the pathogenesis and development of LPR.

Combination of Geniposide and Eleutheroside B Exerts Antidepressant-like Effect on Lipopolysaccharide-Induced Depression Mice Model / 中国结合医学杂志

OBJECTIVE@#To study the antidepressant-like effect and action mechanism of geniposide and eleutheroside B combination treatment on the lipopolysaccharide (LPS)-induced depression mice model.@*METHODS@#Depression mice model was established by lipopolysaccharide (LPS) injection. Totally 48 mice were randomly divided into 6 groups (8 rats per group) according to a random number table, including normal, model, fluoxetine (20 mg/kg), geniposide (100 mg/kg) + eleutheroside B (100 mg/kg), geniposide + eleutheroside B + WAY 100635 (0.03 mg/kg), geniposide + eleutheroside B+ N-methyl-D-aspartic acid receptor (NMDA, 75 mg/kg) groups, respectively. After continuous administration for 10 days, autonomic activity tests after 30 min of administration were performed on the 10th day. On the 11th day, except for the normal group, the mice in the other groups were intraperitoneally injected with LPS (1 mg/kg), and the behavioral tests were performed 4 h later. Enzyme linked immunosorbent assay was used to detect tumor necrosis factor alpha (TNF- α) and interleukin-1 β (IL-1 β) levels in mice serum. The mRNA expression of indoleamine 2,3-dioxygenase (IDO) and nuclear transcription factor (NF- κB) were detected by real-time quantitative polymerase chain reaction. Western-blot analysis was used to detect IDO and NF- κB protein expressions in hippocampus tissue.@*RESULTS@#Compared with the normal group, a single administration of LPS increased the immobility time in the forced swimming test (FST) and tail suspension test (TST, P<0.01), without affecting autonomous activity. Compared with the model group, fluoxetine and geniposide + eleutheroside B administration significantly improved the immobility time of depressed mice in the FST and TST, decreased serum IL-1 β content, inhibited the expression levels of NF- κ B gene and protein in hippocampus tissues (P<0.05 or P<0.01). Compared with the model group, geniposide + eleutheroside B treatment significantly reduced serum TNF-α content and inhibited IDO mRNA and protein expressions in hippocampus (P<0.05 or P<0.01). In addition, NMDA partly prevented the inhibition of IDO mRNA expression by geniposide + eleutheroside B; NMDA and WAY-100635 also partly prevented the reduction of IL-1 ß content induced by geniposide + eleutheroside B treatment (P<0.05 or P<0.01).@*CONCLUSIONS@#The combination of geniposide and eleutheroside B showed a certain antidepression-like effect. Its main mechanism of action may be contributed to inhibiting the activation of NF- κB, decreasing the proinflammatory cytokines such as TNF-α, IL-1 β, and inhibiting in the neuroinflammatory reaction. Additionally, it also affects tryptophan metabolism, reduces the expression of a key enzyme of tryptophan metabolism, IDO. And this antidepressant-like effect may be mediated by 5-hydroxytryptamine and glutamate systems.

Optimized experimental method for determination of hydroxyproline in lung tissue of mice / 医学研究生学报

ObjectiveThere are many methods for the detection of hydroxyproline (HYP), but few of them are suitable for the detection of lung tissue in mice. We intend to establish an accurate and reliable method for measuring HYP levels based on mouse lung tissues to assess the degree of fibrosis development more effectively.MethodsBased on the alkali hydrolysis method, the effects of the concentration of alkali hydrolysate and hydrolysis time on the determination results of HYP level in mice lung tissue were compared; the effects of the changes of experimental conditions on the determination results of HYP standard were compared; and the results of the determination of HYP level in mice lung tissue under dry and wet conditions were compared on the basis of the above experimental results.ResultsThe optimum concentration of alkali hydrolysate is 2 mol/L and the optimum hydrolysis time is 20 min. The optimum pH value of citric acid buffer is 6.0-6.5. The optimum solvent for chloramine T is methanol, the optimum reaction time for chloramine T solution is 15 min, the optimum reaction time for perchloric acid solution is 5 min, and the optimum reaction time for 4-(dimethylamino) benzoyl toluene is 5 min. The optimum condition of aldehyde solution color development is that it is bathed in water at 85 for 3 minutes. Some related reagents are stored in suitable environment after preparation, and the experimental data will not be affected within 7 days. Dry lung tissue of mice can improve the detection level of HYP. The improved experimental protocol was applied to the bleomycin-induced mouse pulmonary fibrosis model, and the HYP measurement results were significantly higher than that of the original protocol.ConclusionAn accurate and reliable method for the determination of hydroxyproline in lung tissue of mice was established.

Program Design of Motor Imagery with Regular Rhythm / 中国康复理论与实践

Objective:To propose a motor imagery paradigm with regular rhythm to improve the accuracy of classification in brain-computer interface based on motor imagery. Methods:Ten untrained healthy subjects were asked to image the movement guided by the picture accompanied with regular rhythm. The common spatial pattern feature extraction algorithm and Fisher classifier were used for classification, and compared with the traditional paradigm and the motor imagery paradigm accompanying with irregular rhythm. Results:More desynchronization was found as motor imagery with regular rhythm, while the classification accuracy improved. Conclusion:Motor imagery with regular rhythm gives a new idea for active rehabilitation training based on brain-computer interface.

Program Design of Motor Imagery with Regular Rhythm / 中国康复理论与实践

Objective:To propose a motor imagery paradigm with regular rhythm to improve the accuracy of classification in brain-computer interface based on motor imagery. Methods:Ten untrained healthy subjects were asked to image the movement guided by the picture accompanied with regular rhythm. The common spatial pattern feature extraction algorithm and Fisher classifier were used for classification, and compared with the traditional paradigm and the motor imagery paradigm accompanying with irregular rhythm. Results:More desynchronization was found as motor imagery with regular rhythm, while the classification accuracy improved. Conclusion:Motor imagery with regular rhythm gives a new idea for active rehabilitation training based on brain-computer interface.

DREAM: a multifunctional transcriptional regulator / 生理学报

DREAM (downstream regulatory element antagonist modulator), Calsenilin and KChIP3 (potassium channel interacting protein 3) belong to the neuronal calcium sensor (NCS) superfamily, which transduces the intracellular calcium signaling into a variety of activities. They are encoded by the same gene locus, but have distinct subcellular locations. DREAM was first found to interact with DRE (downstream regulatory element) site in the vicinity of the promoter of prodynorphin gene to suppress gene transcription. Calcium can disassemble this interaction by binding reversibly to DREAM protein on its four EF-hand motifs. Apart from having calcium dependent DRE site binding, DREAM can also interact with other transcription factors, such as cAMP responsive element binding protein (CREB), CREB-binding protein (CBP) and cAMP responsive element modulator (CREM), by this concerted actions, DREAM extends the gene pool under its control. DREAM is predominantly expressed in central nervous system with its highest level in cerebellum, and accumulating evidence demonstrated that DREAM might play important roles in pain sensitivity. Novel findings have shown that DREAM is also involved in learning and memory processes, Alzheimer's disease and stroke. This mini-review provides a brief introduction of its discovery history and protein structure properties, focusing on the mechanism of DREAM nuclear translocation and gene transcription regulation functions.

Gonioscopy and ultrasound biomicroscopy in the detection of angle closure in patients with shallow anterior chamber / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To assess the agreement between gonioscopy and ultrasound biomicroscopy (UBM) in detecting angle closure in Chinese patients with shallow anterior chamber.</p><p><b>METHODS</b>An observational comparative study of the two different examination methods was conducted. Patients with normal intraocular pressure and temporal peripheral anterior chamber depth less than a quarter of corneal thickness based on slit lamp examination were included in this study from December 2007 to May 2009 in the outpatient clinic of First Hospital of Tsinghua University. Gonioscopy was performed with a Goldman goniolens in dark room first and followed by full beam light and indentation. If the filtering trabecular meshwork was invisible or any peripheral anterior synechia was found, that quadrant of the angle was considered closed. UBM was first undertaken in a darkened room then repeated with normal room lighting. If iridotrabecular apposition was showed, that quadrant of the angle was considered closed. The status of angle closure of each quadrant with different methods was recorded.</p><p><b>RESULTS</b>85 eyes of 46 patients were included in this study. The agreement between gonioscopy and UBM was poor (Κ<0.4) with Kappa analysis in both dark and light conditions in each quadrant. The accordance of agreement between gonioscopy and UBM was hardly affected by age or sex, while in dark condition, eyes with deeper anterior chamber (P=0.005) or plateau iris configuration tended to produce different results (P=0.075) in the 2 methods.</p><p><b>CONCLUSION</b>Gonioscopy and UBM are both indispensable methods for detecting angle closure, neither can completely replace the other.</p>

Construction and preliminary applications of a Saccharomyces cerevisiae detection plasmid using for screening promoter elements / 药学学报

Synthetic biology of natural products is the design and construction of new biological systems by transferring a metabolic pathway of interest products into a chassis. Large-scale production of natural products is achieved by coordinate expression of multiple genes involved in genetic pathway of desired products. Promoters are cis-elements and play important roles in the balance of the metabolic pathways controlled by multiple genes by regulating gene expression. A detection plasmid of Saccharomyces cerevisiae was constructed based on DsRed-Monomer gene encoding for a red fluorescent protein. This plasmid was used for screening the efficient promoters applying for multiple gene-controlled pathways. First of all, eight pairs of primers specific to DsRed-Monomer gene were synthesized. The rapid cloning of DsRed-Monomer gene was performed based on step-by-step extension of a short region of the gene through a series of PCR reactions. All cloned sequences were confirmed by DNA sequencing. A vector named pEASYDs-M containing full-length DsRed-Monomer gene was constructed and was used as the template for the construction of S. cerevisiae expression vector named for pYeDP60-Ds-M. pYeDP60-Ds-M was then transformed into S. cerevisiae for heterologous expression of DsRed-Monomer gene. SDS-PAGE, Western blot and fluorescence microscopy results showed that the recombinant DsRed-Monomer protein was expressed successfully in S. cerevisiae. The well-characterized DsRed-Monomer gene was then cloned into a yeast expression vector pGBT9 to obtain a promoter detection plasmid pGBT9Red. For determination efficacy of pGBT9Red, six promoters (including four inducible promoters and two constitutive promoters) were cloned by PCR from the S. cerevisiae genome, and cloned into pGBT9Red by placing upstream of DsRed-Monomer gene, separately. The fluorescence microscopy results indicated that the six promoters (GAL1, GAL2, GAL7, GAL10, TEF2 and PGK1) can regulate the expression of DsRed-Monomer gene. The successful construction of pGBT9Red lays the foundation for further analysis of promoter activity and screening of promoter element libraries.

IL-1β promotes the hematopoietic support of human umbilical cord mesenchymal stem cells / 中国实验血液学杂志

This study was aimed to investigate the effect of IL-1β on hematopoietic support of human umbilical cord mesenchymal stem cells (hUC-MSC). 2×10(6) hUC-MSC were seeded in 75 cm(2) flasks, after adherence to wall for 2 h, 10 ng/ml IL-1β was added in hUC-MSC supernatant and cultured for 36 h, then the culture supernatants and cells were harvested. The effect of conditioned medium with/without IL-1β on CD34(+) cell hematopoietic support was observed, mRNA expression changes of hUC-MSC cultured in medium with/without IL-1β were monitored by real time PCR, the differences in hematopoiesis-related factors were detected by ELISA. The results showed that the conditioned culture medium of hUC-MSC with IL-1β enhanced the ability to form colony of CD34(+) cells, especially CFU-G and CFU-GM in vitro; IL-1β promoted the mRNA expression of GM-CSF, G-CSF, IL-6 on MSC; IL-1β also promoted the secretion of GM-CSF, G-CSF, and IL-6 protein from hUC-MSC. It is concluded that IL-1β enhances hematopoietic support capacity especially, capability of MSC to myeloid differentiation.

Supportive effects of conditioned culture media of human umbilical cord mesenchymal stem cells on hematopoiesis in vitro / 中国实验血液学杂志

This study was aimed to explore whether the conditioned culture medium of human umbilical cord-derived mesenchymal stem cells (hUC-MSC) has supportive effects on hematopoiesis in vitro. hUC-MSC were cultured in 75 cm(2) culture flasks at a concentration of 2×10(6) cells per flask. After 48 h, the conditioned culture medium was harvested. CD34(+) cells were isolated with the human cord blood CD34 positive selection kit. The CD34(+) cells were plated in three different culture systems: the culture supernatant from hUC-MSC added into incomplete methylcellulose without recombinant human cytokines as conditioned culture medium; the complete methylcellulose medium with recombinant human cytokines as positive control medium; incomplete methylcellulose adding DMEM/F12 with 10% FBS instead of conditioned culture medium as the negative control medium. After 14 days of culture, colonies containing ≥ 50 cells were scored and types of colonies were classified under inverted microscope. The immunophenotypes of cells which were collected from the colonies were detected by flow cytometry. The results showed that conditioned culture medium of hUC-MSC supported the differentiation of CD34(+) cells into CFU-G (47.67 ± 0.58), CFU-GM (48.67 ± 4.73) and CFU-M (3.00 ± 2.00) in vitro, while the CFU-E, BFU-E or CFU-GEMM were absent. Comparatively, in the positive control medium all kinds of CFU were observed. Interestingly, the percentage of CD45(+)cells of CFU in conditioned culture medium (97.43 ± 2.15)% was more than CD45(+)cells in positive control medium (39.69 ± 0.96)% (P < 0.05). It is concluded that the conditioned culture medium of hUC-MSC has been confirmed to have ability to support hematopoiesis separately in vitro. Besides, it enhances the differentiation of CD34(+) cells into myeloid cells except cells of erythroid lineage.

Development and evaluation of a safe and effective sugar-free herbal lollipop that kills cavity-causing bacteria / 国际口腔科学杂志·英文版

Dental caries (tooth decay) is caused by a specific group of cariogenic bacteria, like Streptococcus mutans, which convert dietary sugars into acids that dissolve the mineral in tooth structure. Killing cariogenic bacteria is an effective way to control or prevent tooth decay. In a previous study, we discovered a novel compound (Glycyrrhizol A), from the extraction of licorice roots, with strong antimicrobial activity against cariogenic bacteria. In the current study, we developed a method to produce these specific herbal extracts in large quantities, and then used these extracts to develop a sugar-free lollipop that effectively kills cariogenic bacteria like Streptococcus mutans. Further studies showed that these sugar-free lollipops are safe and their antimicrobial activity is stable. Two pilot human studies indicate that a brief application of these lollipops (twice a day for ten days) led to a marked reduction of cariogenic bacteria in oral cavity among most human subjects tested. This herbal lollipop could be a novel tool to promote oral health through functional foods.

Soy isoflavone and its effect to regulate hypothalamus and peripheral orexigenic gene expression in ovariectomized rats fed on a high-fat diet / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To explore the effect of soy isoflavone on obesity in the light of hypothalamus and peripheral orexigenic gene regulation.</p><p><b>METHODS</b>Fifty-four female rats were randomly assigned to 6 groups: one sham-operated group (SHAM), one ovariectomized (OVX) control group, three OVX groups fed with 400 ppm (L-SI), 1200 ppm (M-SI) and 3600 ppm (H-SI) isoflavone respectively, and one OVX group receiving 0.45 ppm diethylstilbestrol (EC). All rats were allowed to take high-fat diet for 4 weeks. Some neuropeptides were measured by RT-PCR. These neuropeptides included NPY, pro-opiomelanocortin (POMC), cocaine and amphetamine regulated transcript (CART), orexin, melanin-concentrating hormone (MCH), melanin-concentrating hormone precursor (P-MCH), ghrelin, and leptin.</p><p><b>RESULTS</b>Compared with the OVX control group, the body weight and food intake in the H-SI group were reduced significantly and there was a significant dose-dependent manner in the 3 isoflavone groups. The results of RT-PCR showed that the NPY level in the 3 isoflavone groups was significantly increased and the POMC/CART gene expression decreased significantly in rats' hypothalamus compared with that in the OVX control group. However, the expression of orexin, MCH and P-MCH had no change. The peripheral grelin mRNA expression was higher in the 3 isoflavone groups, while leptin gene expression in the fat was not consistent.</p><p><b>CONCLUSIONS</b>This research showed that isoflavone could prevent obesity induced by high-fat diet and ovariectomy through regulating hypothalamus and peripheral orexigenic gene expressions associated with food intake.</p>

Targeted antimicrobial therapy against Streptococcus mutans establishes protective non-cariogenic oral biofilms and reduces subsequent infection / 国际口腔科学杂志·英文版

<p><b>AIM</b>Dental biofilms are complex communities composed largely of harmless bacteria. Certain pathogenic species including Streptococcus mutans (S. mutans) can become predominant when host factors such as dietary sucrose intake imbalance the biofilm ecology. Current approaches to control S. mutans infection are not pathogen-specific and eliminate the entire oral community along with any protective benefits provided. Here, we tested the hypothesis that removal of S. mutans from the oral community through targeted antimicrobial therapy achieves protection against subsequent S. mutans colonization.</p><p><b>METHODOLOGY</b>Controlled amounts of S. mutans were mixed with S. mutans-free saliva, grown into biofilms and visualized by antibody staining and cfu quantization. Two specifically-targeted antimicrobial peptides (STAMPs) against S. mutans were tested for their ability to reduce S. mutans biofilm incorporation upon treatment of the inocula. The resulting biofilms were also evaluated for their ability to resist subsequent exogenous S. mutans colonization.</p><p><b>RESULTS</b>S. mutans colonization was considerably reduced ( +/- 0.4 fold reduction, P=0.01) when the surface was preoccupied with saliva-derived biofilms. Furthermore, treatment with S. mutans-specific STAMPs yielded S. mutans-deficient biofilms with significant protection against further S. mutans colonization (5 minutes treatment: 38 +/- 13 fold reduction P=0.01; 16 hours treatment: 96 +/- 28 fold reduction P=0.07).</p><p><b>CONCLUSION</b>S. mutans infection is reduced by the presence of existing biofilms. Thus maintaining a healthy or "normal" biofilm through targeted antimicrobial therapy (such as the STAMPs) could represent an effective strategy for the treatment and prevention of S. mutans colonization in the oral cavity and caries progression.</p>

Analysis of environmental factors and polycystic ovary syndrome / 中华妇产科杂志

Objective To investigate environmental risk factors associated with polycystic ovary syndrome(PCOS).Methods A retrospective study was carried out from Feb 2005 to Apr 2006.A total of 108 cases with PCOS and 108 patients without PCOS(control group)were interviewed using a designed questionnaire.Results Univariate analysis of environmental factors indicated that risk factors related to PCOS were:occupation,education,disposable plastic cup for drinking,cooking oil fume and indoor decoration,all of which were significantly related to PCOS(P

Breeding of Arachidonic Acid Producting Strain with Mortierella Sabellina by Ultraviolet Mutation / 微生物学通报

In order to gain the strain which has high growth vigor and whose biomass concentration satisfy further fermentation to produce arachidonic acid, we took the yield of microbes olein and arachidonic acid as evaluative index, adopted twice ultraviolet mutation, determined the time of ultraviolet irradiation by single factor expriment, and then the content of the arachidonic acid was measured by GC. The results of this experiment showed that the power of viltalight lamp was 20 W, exposure distance was 30 cm and exposure time was 80 s, lethality of spores of Mortierella isabellina was 76.4%. After twice ultraviolet mutation and repeatedly screen, a mutation of Mortierella isabellina Z80s2-109 whose arachidonic acid concentration in biomass were 3.77 times of the control strains was obtained and genetic stability.

Analysis on Measures of Enhancing Quality Control of Pharmacy Services in Hospitals / 中国药房

OBJECTIVE:To study the effectiveness of the management measures on the enhancing of pharmacy services quality in our hospital.METHODS:The concerned data about the management measures such as rules that‘those who rece_ ived the prescriptions the first should be responsible for which’,activity that‘no error,no appealing for100days’,rules th_ at‘exchanging notebooks about working quality’,etc.,were analyzed by SPSS10.0software package.RESULTS:The ef?fective practice of management measures can increase patients'satisfaction and trust in the consultancy.CONCLUSION:The management measures are helpful in the enhancement of pharmacy services quality and the information communication between pharmacy department and the other concerned departments.

Preparation and identification of monoclonal antibodies against cell receptors of acute hemorrhagic conjunctivitis viruses / 中国免疫学杂志

To prepare monoclonal antibodies for blocking acute hemorrhagic conjunctivitis (AHC)viruses,BALB/C mice were immunized with Hela cells.The spleen of immunemouse was re-moved,and spleen cells were fused with SP2/0 cells.The antibodies against cell receptors wereselected by cell protection assay.Four cell strains secreted McAbs designated R7.R8.R16 andR24 were screened to be able to block cellular receptors and inhibit the infection of Ev70 as wellas CA24v on Hela cell.These results demonstrate that the receptors for Ev70 and CA24v are si-miliar.Indirect immunofluorescence test with the four McAbs showed definite fluorescence onliving Hela cells,which the intensity was concordance with the mean intensity resulted fromflowcytometer.Positive reaction on fixed Hela cells were only observed with R7 and R8.