Combating the SARS-CoV-2 Omicron variant with non-Omicron neutralizing antibodies

The highly mutated and transmissible Omicron variant has provoked serious concerns over its decreased sensitivity to the current coronavirus disease 2019 (COVID-19) vaccines and evasion from most anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) neutralizing antibodies (NAbs). In this study, we explored the possibility of combatting the Omicron variant by constructing bispecific antibodies based on non-Omicron NAbs. We engineered ten IgG-like bispecific antibodies with non-Omicron NAbs named GW01, 16L9, 4L12, and REGN10987 by fusing the single-chain variable fragments (scFvs) of two antibodies through a linker and then connecting them to the Fc region of IgG1. Surprisingly, eight out of ten bispecific antibodies showed high binding affinity to the Omicron receptor-binding domain (RBD) and exhibited extreme breadth and potency against pseudotyped SARS-CoV-2 variants of concern (VOCs) including Omicron, as well as authentic Omicron(+R346K) variants. Six bispecific antibodies containing the cross-NAb GW01 neutralized Omicron variant and retained their abilities to neutralize other sarbecoviruses. Bispecific antibodies inhibited Omicron infection by binding to the ACE2 binding site. A cryo-electron microscopy (cryo-EM) structure study of the representative bispecific antibody FD01 in complex with the Omicron spike (S) revealed 5 distinct trimers and one unique bi-trimer conformation. The structure and mapping analyses of 34 Omicron S variant single mutants elucidated that two scFvs of the bispecific antibody synergistically induced the RBD-down conformation into 3-RBD-up conformation, enlarged the interface area, accommodated the S371L mutation, improved the affinity between a single IgG and the Omicron RBD, and hindered ACE2 binding by forming bi-trimer conformation. Our study offers an important foundation for anti-Omicron NAb design. Engineering bispecific antibodies based on non-Omicron NAbs may provide an efficient solution to combat the Omicron variant.

Clinical outcomes of acute pulmonary embolectomy as the first-line treatment for massive and submassive pulmonary embolism: a single-centre study in China.

BACKGROUND: Acute pulmonary embolism (PE) is one of the most critical cardiovascular diseases. PE treatment ranges from anticoagulation, and systemic thrombolysis to surgical embolectomy and catheter embolectomy. Surgical pulmonary embolectmy (SPE) indications and outcomes are still controversial. Although there have been more favourable SPE reports over the past decades, SPE has not yet been considered broadly as an initial PE therapy and is still considered as a reserve or rescue treatment for acute massive PE when systemic thrombolysis fails. This study aimed to evaluate the early and midterm outcomes of SPE, which was a first-line therapy for acute central major PE in one Chinese single centre. METHODS: A retrospective review of patients who underwent SPE for acute PE was conducted.Patients with chronic thrombus or who underwent thromboendarterectomy were excluded. SPE risk factors for morbidity and mortality were reviewed, and echocardiographic examination were conducted for follow-up studies to access right ventricular function. RESULTS: Overall, 41 patients were included; 17 (41.5%) had submassive PE, and 24 (58.5%) had massive PE. Mean cardiopulmonary bypass time was 103.2 ± 48.9 min, and 10 patients (24.4%) underwent procedures without aortic cross-clamping. Ventilatory support time was 78 h (range, 40-336 h), intensive care unit stay was 7 days (range, 3-13 days), and hospital stay was 16 days (range, 12-23 days). Operative mortalities occurred in 3 massive PE patients, and no mortality occurred in submassive PE patients. The overall SPE mortality rate was 7.31% (3/41). If two systemic thrombolysis cases were excluded, SPE mortality was low (2.56%,1/39), evenlthough there were 2 cases of cardiac arrest preoperatively. Patients' right ventricle function improved postoperatively in follow-ups.There were no deaths related to recurrent PE and chronic pulmonary hypertension in follow-ups, though 3 patients died of cerebral intracranial bleeding, gastric cancer,and brain cancer at 1 year, 3 years, and 8 years postoperatively, respectively. CONCLUSIONS: SPE presented with a low mortality rate when rendered as a first-line treatment in selected massive and submassive acute PE patients. Favorable outcomes of right ventricle function were also observed in the follow-ups. SPE should play the same role as ST in algorithmic acute PE treatment.

Antibody-dependent enhancement (ADE) of SARS-CoV-2 infection in recovered COVID-19 patients: studies based on cellular and structural biology analysis

Antibody-dependent enhancement (ADE) has been reported in several virus infections including dengue fever virus, severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) coronavirus infection. To study whether ADE is involved in COVID-19 infections, in vitro pseudotyped SARS-CoV-2 entry into Raji cells, K562 cells, and primary B cells mediated by plasma from recovered COVID-19 patients were employed as models. The enhancement of SARS-CoV-2 entry into cells was more commonly detected in plasma from severely-affected elderly patients with high titers of SARS-CoV-2 spike protein-specific antibodies. Cellular entry was mediated via the engagement of Fc{gamma}RII receptor through virus-cell membrane fusion, but not by endocytosis. Peptide array scanning analyses showed that antibodies which promote SARS-CoV-2 infection targeted the variable regions of the RBD domain. To further characterize the association between the spike-specific antibody and ADE, an RBD-specific monoclonal antibody (7F3) was isolated from a recovered patient, which potently inhibited SARS-Cov-2 infection of ACE-2 expressing cells and also mediated ADE in Raji cells. Site-directed mutagenesis the spike RBD domain reduced the neutralization activity of 7F3, but did not abolish its binding to the RBD domain. Structural analysis using cryo-electron microscopy (Cryo-EM) revealed that 7F3 binds to spike proteins at a shift-angled pattern with one "up" and two "down" RBDs, resulting in partial overlapping with the receptor binding motif (RBM), while a neutralizing monoclonal antibody that lacked ADE activity binds to spike proteins with three "up" RBDs, resulting in complete overlapping with RBM. Our results revealed that ADE mediated by SARS-CoV-2 spike-specific antibodies could result from binding to the receptor in slightly different pattern from antibodies mediating neutralizations. Studies on ADE using antibodies from recovered patients via cell biology and structural biology technology could be of use for developing novel therapeutic and preventive measures for control of COVID-19 infection.

Neutralizing antibody responses to SARS-CoV-2 in a COVID-19 recovered patient cohort and their implications

BackgroundThe COVID-19 pandemic caused by SARS-CoV-2 coronavirus threatens global public health. Currently, neutralizing antibodies (NAbs) versus this virus are expected to correlate with recovery and protection of this disease. However, the characteristics of these antibodies have not been well studied in association with the clinical manifestations in patients. MethodsPlasma collected from 175 COVID-19 recovered patients with mild symptoms were screened using a safe and sensitive pseudotyped-lentiviral-vector-based neutralization assay. Spike-binding antibody in plasma were determined by ELISA using RBD, S1, and S2 proteins of SARS-CoV-2. The levels and the time course of SARS-CoV-2-specific NAbs and the spike-binding antibodies were monitored at the same time. FindingsSARS-CoV-2 NAbs were unable to cross-reactive with SARS-CoV virus. SARS-CoV-2-specific NAbs were detected in patients from day 10-15 after the onset of the disease and remained thereafter. The titers of NAb among these patients correlated with the spike-binding antibodies targeting S1, RBD, and S2 regions. The titers of NAbs were variable in different patients. Elderly and middle-age patients had significantly higher plasma NAb titers (P<0.0001) and spike-binding antibodies (P=0.0003) than young patients. Notably, among these patients, there were ten patients whose NAb titers were under the detectable level of our assay (ID50: < 40); while in contrast, two patients, showed very high titers of NAb, with ID50 :15989 and 21567 respectively. The NAb titers were positive correlated with plasma CRP levels but negative correlated with the lymphocyte counts of patients at the time of admission, indicating an association between humoral response and cellular immune response. InterpretationThe variations of SARS-CoV-2 specific NAbs in recovered COVID-19 patients may raise the concern about the role of NAbs on disease progression. The correlation of NAb titers with age, lymphocyte counts, and blood CRP levels suggested that the interplay between virus and host immune response in coronavirus infections should be further explored for the development of effective vaccine against SARS-CoV-2 virus. Furthermore, titration of NAb is helpful prior to the use of convalescent plasma for prevention or treatment. FundingMinistry of Science and Technology of China, National Natural Science Foundation of China, Shanghai Municipal Health Commission, and Chinese Academy of Medical Sciences

Effects of Wuzhi Capsules on in vivo Pharmacokinetic Behavior of Apatinib Mesylate in Rats / 中国药房

OBJECTIVE： To establish a method for the concentration determination of apatinib mesylate in plasma of rats， and to investigate the effects of single and multiple administration of Wuzhi capsules on the pharmacokinetic behavior of apatinib mesylate in rats. METHODS：  LC-MS/MS method was used to detect the plasma concentration of apatinib mesylate in rats. Using carbamazepine as internal standard， the determination was performed on Waters XBridge BEH C18 column with mobile phase consisted acetonitrile-0.1％ formic acid solution （45 ∶ 55，V/V） at the flow rate of 0.3 mL/min. The column temperature was 40 ℃. The temperature of injector was 15 ℃， and the sample size was 2 μL. ESI was used for positive ion scanning in MRM mode. The ion pairs m/z used for quantitative analysis were 398.1→212.1 （apatinib mesylate） and  237.2→194.2 （internal standard）. The rats were randomly divided into control group Ⅰ， observation group Ⅰ， control group Ⅱ， observation group Ⅱ， with 6 rats in each group. Control group Ⅰ were given single administration of apatinib mesylate intragastrically （50 mg/kg， similarly hereinafter）. Observation group Ⅰ was given Wuzhi capsules intragastrically （450 mg/kg， similarly hereinafter）， and then 10 min later given apatinib mesylate intragastrically. Control group Ⅱ was given normal saline intragastrically， once a day， for consecutive 7 d， and then were given single administration of apatinib mesylate. Observation group Ⅱ was given Wuzhi capsules intragastrically， once a day， for consecutive 7 d， and then 10 min later were given single administration of apatinib mesylate. The blood samples were collected from intraocular canthus vein plexus and determined 0.25， 0.5， 1.0， 2.0， 2.5， 3.0， 4.0， 6.0， 8.0， 12.0， 24.0 h after intragastric administration， respectively. Pharmacokinetic parameters were apatinib mesylate were calculated and compared among those groups by using DAS 2.1 software and t-test. RESULTS： The linear range of apatinib mesylate were 2-2 000 ng/mL. The lower limit of quantitation was 2 ng/mL. RSDs of intra- day and inter-day were all lower than 10％， and the accuracy were 94.93％-104.68％. Matrix effect did not affect the quantitative analysis of the substance to be measured. Compared with control group Ⅰ， cmax， AUC0-24 h and AUC0-∞ of observation group Ⅰ were increased significantly， CLZ was decreased significantly （P＜0.05）. Compared with control group Ⅱ， AUC0-24 h and AUC0-∞ of observation group Ⅱ were increased significantly， and CLZ was decreased significantly （P＜0.05）. Compared with observation group Ⅰ， AUC0-24 h and AUC0-∞ of observation group Ⅱ were decreased significantly （P＜0.05）. CONCLUSIONS： Established LC-MS/MS method is sensitive and specific， and can be used for the concentration determination of apatinib mesylate in plasma of rats. Wuzhi capsules can influence in vivo pharmacokinetic behavior of apatinib mesylate in rats. The effect of multiple administration of Wuzhi capsules is weaker than that of single administration.

Simultaneous Determination of 3 Kinds of Saponins in Marsdenia tenacissima by UPLC-MS/MS / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of tenacissoside A,tenacissoside H and tenacissoside I in Marsdenia tenacissima. METHODS:UPLC-MS/MS method was adopted. The determination was performed on a Phenomenex Kinetex XB-C18column with mobile phase consisted of 0.1% formic acid solution-acetonitrile(gradient elution)at the flow rate of 0.2 mL/min. The column temperature was set at 40 ℃,and sample size was 5 μ L. Multiple reaction monitoring (MRM)mode was adopted with electrospray ion source as ion source,using positive ion scanning. Source jet voltage was 5 500 V,nebulizer pressure was 60 psi,heating pressure was 60 psi,curtain pressure was 20 psi and cone temp was set at 600 ℃. RESULTS:The linear ranges of tenacissoside A,tenacissoside H and tenacissoside I were 0.1-10 ng/mL(r=0.999 7),0.025-10 ng/mL(r=0.999 5),0.025-10 ng/mL(r=0.998 9),respectively;limited of quantation were 0.1,0.025,0.025 ng/mL,limited of detection were 0.05,0.012 5,0.012 5 ng/mL,respectively;RSDs of precision,stability and reproducibility tests were<4.0%. The recoveries were 97.67%-99.00%(RSD=0.47%,n=6),95.00%-101.67%(RSD=2.59%,n=6),96.67%-103.33%(RSD=2.83%, n=6). CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for simultaneous determination of tenacissoside A,tenacissoside H and tenacissoside I in M.tenacissima.

[Effects of geranylgeranyltransferaseâ silencing on the proliferation of tongue squamous cancer cells].

Objective This study aims to investigate the effect of geranylgeranyltransferaseⅠ (GGTase-Ⅰ) on the proliferation and growth of tongue squamous cancer cells. Methods Three small interfering RNAs (siRNAs) were designed on the basis of the GGTase-Ⅰ sequence in GeneBank. These siRNAs were then transfected into tongue squamous cancer cells Cal-27. The mRNA and protein expression of GGTase-Ⅰ and RhoA were examined by real-time quantitative polymerase chain reaction and Western blotting, respectively. The expression of Cyclin D1 and p21 were examined by Western blotting. The proliferation and growth ability were analyzed by cell counting kit-8 assay and flow cytometry. Results The mRNA and protein expression of GGTase-Ⅰ in Cal-27 was reduced significantly after the GGTase-Ⅰ siRNAs were transfected (P<0.05). No significant difference in RhoA mRNA and protein expression was detected (P>0.05). Cyclin D1 expression decreased, whereas p21 expression increased significantly. The cell cycle was altered, and the growth-proliferative activity was inhibited (P<0.05). Conclusion GGTase-Ⅰ siRNA can inhibit the expression of GGTase-Ⅰ and the proliferative activity of tongue squamous cancer cells. GGTase-Ⅰ may be a potential target for gene therapy in tongue squamous cell cancer.

[Effects of geranylgeranyltransferase â gene silencing by RNA interference on the migration and invasion of tongue carcinoma].

OBJECTIVE: RNA interference was used to silence geranylgeranyltransferase Ⅰ(GGTase-Ⅰ) in vitro and to study the effect of GGTase-Ⅰ on the migration and invasion of tongue squamous cancer cells. METHODS: Three small interfering RNAs (siRNA) were designed according to the GGTase-Ⅰ sequence by Genebank and were transfected into tongue squamous cancer cells Cal-27 to knock down GGTase-Ⅰ expression. The tested cells were divided into three groups, as follows: the RNA-interfered groups (GGTase-Ⅰ siRNA1, GGTase-Ⅰ siRNA 2, GGTase-Ⅰ siRNA 3), a negative control group (disrupted by random sequence NC-siRNA), and a blank control group. GGTase-Ⅰ and RhoA gene expressions were examined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The optimum interference group was screened by qRT-PCR and Western blot and was assigned as the experimental group. Matrix metalloproteinase (MMP)-2 and MMP-9 protein expressions were examined by Western blot. GTP-RhoA expression of protein was examined by GST-pull down. The migration and invasion abilities were analyzed by wound healing assay and Transwell motility assay. RESULTS: GGTase-Ⅰ mRNA and protein expression in Cal-27 decreased significantly after transfection of GGTase-I siRNA (P<0.05). No significant difference of RhoA gene expression was detected. MMP-2, MMP-9, and GTP-RhoA protein expressions decreased significantly (P<0.05). The migration and invasion abilities were inhibited (P<0.05). CONCLUSIONS: To inhibit GGTase-Ⅰ expression, the migration and invasion abilities of tongue squamous cancer cells should also be inhibited. Further studies on GGTase-Ⅰ may provide novel effective molecular targets for tongue squamous cancer cells.

Bibliometric Analysis of Metabolomics / 中国药房

OBJECTIVE:To investigate the present situation of domestic and foreign metabolomics study,and to preliminarily discuss research hotspots and development rules. METHODS:By using bibliometric methods and GoPubMed literature analysis tool,usingMetabolomicsas subject,all literatures were retrieved from PubMed database up to Jul. 31,2016. Those literatures were ranked and analyzed in respects of publication amount,countries and cities,journal sources,research topics,core authors and authors collaborative networks. UsingMetabolomicsMetabonomicsas subjects,all literatures were retrieved from Web of Science database up to Aug. 1,2016. The literatures with high citation frequency were analyzed during 2007-2016 by using the sort-ing function of citation frequency. RESULTS:A total of 15449 domestic and foreign metabolomics literatures were retrieved. The researchers of 2189 literatures came from China,and the amount of published literatures showed a rising trend. 15449 literatures were distributed in 129 countries and regions;10 countries,such as the United States,China,Britain and Germany,were the core countries,and the total number of literatures issued was 12847(83.16%). The United States held the largest share of world publi-cations(4288 literatures,27.76%),and the following was China in order(2189 literatures,14.17%). The city with the largest amount of publications was London(467 literatures),followed by Shanghai,Beijing. A total of 2168 periodicals were involved, and there were 7.1 published literatures averagely. All of the top 20 periodicals were from the United States and European countries (4377 literatures,28.33%). The first 3 subjects with the highest frequency of occurrence were metabolomics,metabolome and me-tabolism;related researches mainly focused on metabolic processes,metabolic networks and pathways,biological markers,pro-teome and genome;main research methods included spectrum analysis and magnetic resonance spectroscopy. Main 6 groups of au-thors were involved,mainly Nicholoson JK,Holmes E,Lindon J and other researchers. None of Chinese researchers had been found among the core authors and author groups. The literatures with high citation frequency mainly focused on the association of metabolism with disease,metabolomics database,metabolomics research methods and biological markers related to disease predic-tion. CONCLUSIONS:Metabolomics has aroused worldwide interest among researchers,and metabolic pathways and biological markers are the focuses in this field. Our researchers have published a large amount of literatures on metabolomics,but there are not high quality periodical carrier or enough cooperation between researchers. It is suggested to enhance the cooperation between various research institutions or grasp the frontier and hotspots of the research in this field so as to push forward the development di-versification and depth of metabolomics research in China.

Study on Compatible Stability of Xiao'aiping Injection Combined with 3 Kinds of Injections / 中国药房

OBJECTIVE:To investigate the compatible stability of Xiao'aiping injection combined with 3 kinds of common in-jections. METHODS:Referring to package inserts,Xiao'aiping injection 40 mL was compatible with 5% Glucose injection,10%Glucose injection or 0.9% Sodium chloride injection 160 mL,respectively. At room temperature(about 25 ℃)and high tempera-ture(40 ℃),the appearance of mixtures were observed at 0,1,2,4,8,12,24,48 h;pH value and the number of insoluble particles were detected. The contents of tenacissoside A and tenacissoside Ⅰ in mixtures were determined by HPLC. RESULTS:Un-der above condition,the mixtures were brownish yellow liquid within 48 h after Xiao'aiping injection was compatible with 5%Glucose injection or 10% Glucose injection;24 h after mixed with 0.9% Sodium chloride injection,the mixture changed from brownish yellow to reddish brown,but no precipitation was found. The pH value of mixtures had no significant change(RSD<1%,n=8). The number of particles ≥25 μm was in line with the requirements of Chinese Pharmacopeia(2015 edition). For-ty-eight hours after mixing,the number of particles ≥10 μm in the mixtures exceeded the pharmacopoeia limits. Within 48 h after mixing,the relative contents of tenacissoside A and tenacissoside I in mixtures had no significant change(RSD<2%,n=8). CON-CLUSIONS:The mixture should be used up within 24 h after Xiao'aiping injection combined with 5% Glucose injection,10%Glucose injection or 0.9% Sodium chloride injection.

Triptolide ameliorates transplant arteriosclerosis in rat aorta allograft by inhibiting the expression of NO synthase / 中华器官移植杂志

Objective To understand the pathological role of nitric oxide synthase 2 (NOS2) in rat allograft vascular lesions and the effects of triptolide.Methods The abdominal aorta transplantation between Wistar and SD rats was used as an animal model.Three groups were set up..the same genome group (group A),the allogeneic control group (group B),and the isogene Triptolide group (group C).The grafts were removed at 7th,28th,and 56th day after surgery.The transplant artery intima thickness was measured.The irnmunohistochemical staining was applied to observe the NOS2 expression in the vascular tissue layers.The integral optical density value in each group was calculated.Results The arterial intima of transplants at 28th and 56th day postoperation was thickened,and that was thickest in group C among the three groups (P＜0.05).There was significant difference in intima thickness and integral optical density between group C with groups A and B (P＜ 0.05).The expression of NOS2 was strongest in group B,and that in group C was significantly weaker than that in group B (P ＜ 0.05).Conclusion Triptolide is capable of slowing down the progression of graft vascular disease by inhibiting the expression of NOS2.

Effects of geranylgeranyltransferaseⅠsilencing on the proliferation of tongue squamous cancer cells / 华西口腔医学杂志

Objective This study aims to investigate the effect of geranylgeranyltransferaseⅠ (GGTase-Ⅰ) on the proliferation and growth of tongue squamous cancer cells. Methods Three small interfering RNAs (siRNAs) were designed on the basis of the GGTase-Ⅰ sequence in GeneBank. These siRNAs were then transfected into tongue squamous cancer cells Cal-27. The mRNA and protein expression of GGTase-Ⅰ and RhoA were examined by real-time quantitative polymerase chain reaction and Western blotting, respectively. The expression of Cyclin D1 and p21 were examined by Western blotting. The proliferation and growth ability were analyzed by cell counting kit-8 assay and flow cytometry. Results The mRNA and protein expression of GGTase-Ⅰ in Cal-27 was reduced significantly after the GGTase-Ⅰ siRNAs were transfected (P<0.05). No significant difference in RhoA mRNA and protein expression was detected (P>0.05). Cyclin D1 expression decreased, whereas p21 expression increased significantly. The cell cycle was altered, and the growth-proliferative activity was inhibited (P<0.05). Conclusion GGTase-Ⅰ siRNA can inhibit the expression of GGTase-Ⅰ and the proliferative activity of tongue squamous cancer cells. GGTase-Ⅰ may be a potential target for gene therapy in tongue squamous cell cancer.

[Effects of RhoA silencing on proliferation of tongue squamous cancer cells].

OBJECTIVE: This study investigated the effect of RhoA silencing through RNA interference on proliferation and growth of tongue cancer cells, as well as explored the possible mechanisms of this effect. METHODS: SSC-4 tongue cancer cells were cultured in vitro and then transfected with small interfering RNA to knock down RhoA expression. The tested cells were divided into three groups: experimental group (experimental group 1: transfected with RhoA-siRNA-1; experi-mental group 2: transfected with RhoA-siRNA-2), negative control group (transfected by random sequence NC-siRNA), and blank control group (transfected with Lipofectamine). The expression levels of RhoA mRNA were respectively measured by quantitative real-time polymerase chain reaction and western blot assay. Moreover, the expression levels of cyclin D1, p21, and p27 and RhoA protein were evaluated by Western blot assay. Proliferation and growth potentiality were analyzed through evaluation of doubling times and methyl thiazolyl tetrazolium assessment. RESULTS: The expression levels of RhoA gene and protein of experimental groups significantly decreased following siRNA transfection compared with those in the negative and blank control groups. The expression of cyclin D1 decreased significantly and that of p21 and p27 increased significantly. The doubling time was extended and the growth potentiality decreased. CONCLUSIONS: The results indicated that RhoA silencing can inhibit proliferation of tongue cancer cells, whereas RhoA affects cell proliferation by regulating the cell cycle pathway. Thus, RhoA is a potential target in gene therapy for tongue cancer.

Effects of RhoA gene silencing by RNA interference on invasion of tongue carcinoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effects of RhoA down-regulation by RNA interference on the invasion of tongue carcinoma Tca8113 and SCC-4.</p><p><b>METHODS</b>Determination of the human RhoA sequence as well as the design and constructionof a short specific small interfering RNAs (siRNA) were performed. The siRNA of RhoA gene was transfected into humantongue squamous cell carcinoma Tca8113 and SCC-4 cells line by Lipofectamine 2000. Quantitative real-time polymerasechain reaction was used to examine the mRNA expressionlevels of RhoA. Protein expressions of mRNA, galectin-3,and matrix metalloproteinase (MMP)-9 were evaluated byWestern blot. Transwell invasion assay was performed toassess the invasion ability of tongue carcinoma.</p><p><b>RESULTS</b>RhoA expressions in Tca8113 and SCC-4 cells were reducedsignificantly after transfection of RhoA-siRNA. Protein levels f galectin-3 and MVP-9 were also down-regulated significantly. Invasion ability was inhibited as well.</p><p><b>CONCLUSION</b>RhoA-siRNA can effectively inhibit RhoA expression in Tca8113 and SCC-4 cells. The invasion ability of tongue carcinoma cells decreased with down-regulation of the protein expressions of galectin-3 and MMP-9, indicating that RhoA-siRNA can inhibit invasion of tongue carcinoma. Results show that RhoA may play an important role in the processes of invasion and metastasis of tongue carcinoma.</p>

Rsearch of mechanism inhibitory effect on MicroRNA218-Robo1 pathway in breast cancer cell migration / 国际外科学杂志

Objective To observe the migration and inhibition mechanism of MicroRNA218-Robo1 pathway for breast cancer.Methods A total of 40 BALB/c-nu/nu female mice were randomly divided into four groups.Each group was transfected over-expression MicroRNA218 MDA-MB-231 breast cancer cells, co-over-expression MicroRNA218 and Robo1 MDA-MB-231 breast cancer cells, knock-down Robo1 MDA-MB-231 breast cancer cells and the control MDA-MB-231 breast cancer cells.The tumor volume was examined every two weeks.Results Tumor volume of MicroRNA218 group was obviously less than control group, tumor volume of Robo1 knock out group was obviously less than common MicroRNA218 high expression and Robo1 group, the difference was statistically significant;MicroRNA218 and Robol knockout group than the control group, the increase in breast cancer cells apoptosis, cell proliferation and angiogenesis is restrained.Conclusions MicroRNA218 inhibited the migration of breast cancer by down-regulating the expression of Robo1.

Clinical evaluation of percutaneous atrial septal defect closer guided by transthoracic echocardiography and/or transesophageal echocardiography / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the safety and efficacy of percutaneous closure of the single secundum atrial septal defects (ASD) guided by transthoracic echocardiography (TTE) and/or transesophageal echocardiography (TEE).
@*METHODS@#From January, 2014 to December, 2014, thirty-two patients with single secundum ASD from Department of Cardiovascular Surgery, Union Hospital, Fujian Medical University, were treated with percutaneous closure of ASD guided by TTE or TEE.
@*RESULTS@#Thirty-two patients underwent ASD closure successfully, except one patient showed trivial residual shunts, which disappeared one month later. The remaining 31 patients were subjected to TTE. At once or at the 1st or 3rd month after the procedure, no ASD migration or residual shunts were observed.
@*CONCLUSION@#Percutaneous closure of ASD guided by TTE or TEE is a safe and effective surgery method with minimal invasion and can avoid the chest incision and radioscopy.

Inhibition of central Na(+)/H(+) exchanger type 3 can alleviate sleep apnea in Sprague-Dawley rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Recent studies showed the central Na+/H+ exchanger type 3 (NHE3) has a close relationship with ventilation control. The objective of the study is to investigate the role of NHE3 in sleep apnea in Sprague-Dawley (SD) rats.</p><p><b>METHODS</b>A sleep study was performed on 20 male SD rats to analyze the correlation between the sleep apneic events and total NHE3 protein content and inactive NHE3(pS552) in the brainstem measured by Western blotting. Another 20 adult male SD rats received 3 days of sleep and respiration monitoring for 6 hours a day, with adaption on the first day, 0.5% DMSO microinjection into the fourth ventricle on the second day, and AVE0657 (specific inhibitor of NHE3) microinjection on the third day. Rats were divided into two groups with injection of 5 µmol/L or 8 µmol/L AVE0657 before the sleep study. The effects of AVE0657 on sleep apnea and sleep structure of rats were analyzed through self-control.</p><p><b>RESULTS</b>The total post-sigh apnea index (TPSAI) and post-sigh apnea index in non-rapid eye movement (NREM) sleep (NPSAI) and total apnea index (AI) in NREM sleep (NAI) were negatively correlated with NHE3(pS552) protein contents in the brainstem (r = -0.534, -0.547 and -0.505, respectively, P < 0.05). The spontaneous apnea index in REM sleep (RSPAI) was positively correlated with the level of NHE3(pS552) protein expression in the brainstem (r = 0.556, P < 0.05). However, the sleep AI had no relationship with total NHE3 protein. Compared with the blank control and microinjection of 0.5% DMSO, 5 µmol/L AVE0657 significantly reduced the total AI and NPSAI (both P < 0.05) without a significant effect on sleep architecture. In contrast to blank control and microinjection of 0.5% DMSO, injection of 8 µmol/L AVE0657 significantly reduced the AI and PSAI in NREM and REM sleep (all P < 0.05).</p><p><b>CONCLUSIONS</b>The severity of sleep apnea was negatively correlated with central inactive NHE3. A specific inhibitor of NHE3 decreased the sleep AI. Thus, our results indicate that central NHE3 might be a molecular target for sleep apnea treatment, whose inhibitors may be potential therapeutic drugs for sleep apnea.</p>

The comparison of the GlideScope and direct laryngoscopy in snoring surgery under general anesthesia endotracheal intubation / 国际外科学杂志

Objective To compare effect of the GlideScope and direct laryngoscopy in snoring surgery.Methods Fourty patients scheduled for orotracheal intubation of general anesthesia in snoring surgery wererandomly divided into GlideScope group ( A group) and direct laryngoscope group ( B group). Two groups were recorded values of heart rate (HR),systolic blood pressure (SBP), diastolic blood pressure (DBP)before induction (T1), after induction (T2), intubation (T3) and 1 (T4), 3 (T5), 5 (T6) min after intubation and plasma norepinephrine (NE) concentrations in T1, T5, T6. Results ( 1 ) Values of HR, SBP and DBP of two groups were compared in T1 and T2, the difference was not significant ( P ＞ 0.05 ); Values of SBP and DBP of two groups compared T1 to T2, and the difference was significant ( P ＜ 0. 05); (2) Values of HR, SBP and DBP of two groups compared in T3, T4 and T5, and the difference was statistically significant (P ＜0.05);(3 ) Values of the concentration of plasma NE of two groups compared in T1 ,T5 and T6, and the difference was statistically significant ( P ＜ 0. 05 ). Conclusion Cardiovascular response and stress response of orotracheal intubation using a videolaryngoscope is lower than using ordinary laryngoscope in snoring surgery.

The significance of hyperhomocysteineamia in heart transplant patients / 中华器官移植杂志

Objective To investigate plasma homocystein (Hcy) level in cardiac transplant patients.Methods Fourteen cases of heart transplant patients were recruited in this study. Plasma Hcy levels were measured by using high-performance liquid chromatography with fluorescence detection and flow-mediated dilatation in brachial arteries detected with high-resolution ultrasound in all patients, while coronary artery angiography were performed in 7 patients.Results In 2 cases of heart transplant who had single coronary artery stenosis, the plasma Hcy levels were above 12 ?mol/L. The average plasma Hcy levels in heart transplant patients were higher than in health volunteers [( 13.47? 2.78) ?mol/L vs (9.26? 3.57) ?mol/L], and flow-mediated dilatation in brachial arteries of heart transplant patients was lower than that of health volunteers [( 8.2? 3.7) % vs ( 12.5? 1.6) %]. There was a linear correlation between levels of plasma Hcy and flow-mediated dilatation (r= -0.804). Conclusion Endothelial dysfunction in the heart transplant patents might be contributed to hyperhomocysteinemia which may be potential cause of transplant coronary artery disease.

AN ELECTRON MICROSCOPIC HISTOCHEMICAL OBSERVATION ON LYSOSOMAL ACID PHOSPHATASE AND ARYLSULPHATASE IN THE HEPATOCYTE OF GUINEA PIG, PIGEON AND TOAD / 解剖学报

By using the cytochemical methods, the activity of lysosomal acid phosphatase (ACP) and arylsulphatase (ArSase) in liver cells of the three kinds of vertebrates, guinea pig, pigeon and toad, was observed. The results show that the distribution of the two enzymes has some difference in the hepatocytes of all three kinds of animals. There are four forms of ArSase localization in the dense bodies of hepatocytes. Furthermore the three kinds of animals have different ArSase localization characteristics. The biological significance of these phenomena was discussed.