Prediction of potential therapeutic drugs against SARS-CoV-2 by using Connectivity Map based on transcriptome data.

OBJECTIVE: Transcriptome data related to severe acute respiratory syndrome-related coronavirus 2 (a novel coronavirus discovered in 2019, SARS-CoV-2) in GEO database were downloaded. Based on the data, influence of SARS-CoV-2 on human cells was analyzed and potential therapeutic compounds against the SARS-CoV-2 were screened. MATERIALS AND METHODS: R package "DESeq2" was used for differential gene analysis on the data of cells infected or non-infected with SARS-CoV-2. The "ClusterProfiler" package was used for GO functional annotation and KEGG pathway enrichment analysis of the differentially expressed genes (DEGs). A protein-protein interaction (PPI) network of the DEGs was constructed through STRING website, and the key subset in the PPI network was identified after visualization by Cytoscape software. Connectivity Map (CMap) database was used to screen known compounds that caused genomic change reverse to that caused by SARS-CoV-2. RESULTS: By intersecting DEGs in two datasets, a total of 145 DEGs were screened out, among which 136 genes were upregulated and 9 genes were downregulated in SARS-CoV-2-infected cells. Functional enrichment analyses revealed that these genes were mainly associated with the pathways involved in viral infection, inflammatory response, and immunity. The CMap research found that there were three compounds with a median_tau_score less than -90, namely triptolide, tivozanib and daunorubicin. CONCLUSIONS: SARS-CoV-2 can cause abnormal changes in a large number of molecules and related signaling pathways in human cells, among which IL-17 and TNF signaling pathways may play a key role in pathogenic process of SARS-CoV-2. Here, three compounds that may be effective for the treatment of SARS-CoV-2 were screened, which would provide new options for improving treatment of patients infected with SARS-CoV-2.

The gene regulatory network in different brain regions of neuropathic pain mouse models.

OBJECTIVE: Neuropathic pain is directly developed from lesions or somatosensory nervous system diseases that are associated with emotion regulation. In general population, the incidence of neuropathic pain ranges from 7% to 10%, but the underlying mechanism remains largely unknown. Neuropathic pain is often associated with structural and functional abnormalities in multiple brain regions, and its regulation has been shown to correspond with the forebrain, including nucleus accumbens (NAc), medial prefrontal cortex (mPFC) and periaqueductal gray (PAG). MATERIALS AND METHODS: To investigate the molecular mechanism of neuropathic pain across different brain regions, we identified the differentially expressed genes (DEGs) between the spared nerve injury model (SNI) mice suffering neuropathic pain and the control Sham mice in NAc, mPFC and PAG three brain regions, and mapped these genes onto a comprehensively functional association network. Thereafter, novel neuropathic pain genes in these three regions were identified using With Random Walk with Restart (RWR) analysis, such as Asic3, Cd200r1 and MT2, besides well-known Capn11 and CYP2E1. RESULTS: Interactions or cross talks among DEGs in NAc, mPFC and PAG three brain regions were discovered. CONCLUSIONS: Our results provide novel insights into neuropathic pain and help to explore therapeutic targets in the treatment.

Clinical significance of overexpressed cyclin-dependent kinase subunits 1 and 2 in esophageal carcinoma.

The mammalian cyclin-dependent kinase subunit (Cks) family has two members, Cks1 and Cks2. Overexpression of Cks1 and Cks2 has been reported to be associated with high aggressiveness and poor prognosis in several malignancies, including prostate and hepatocellular carcinomas. However, whether Cks1 and Cks2 are overexpressed in esophageal carcinoma remains uncharacterized. To investigate whether overexpression of the Cks family is clinically relevant in esophageal carcinoma, and whether expression patterns of Cks1 and Cks2 can serve as biomarkers for esophageal carcinoma. Real-time quantitative reverse transcription polymerase chain reaction, immunohistochemistry, and Western blot analyses were applied to detect the expression of Cks1 and Cks2 at the mRNA and protein levels, respectively. The associations between Cks1 or Cks2 expressions and clinical features and p27(kip1) expressions in esophageal carcinoma were analyzed. Comparing with the adjacent noncancerous tissues, esophageal carcinoma exhibited elevated expression of Cks1 in 58% cases at the mRNA level and 54% cases at the protein level, and elevated expression of Cks2 in 65% cases at the mRNA level and 61% cases at the protein level, respectively. The expressions of both Cks1 and Cks2 were negatively associated with the p27(kip1) protein level in the tumor tissues. Furthermore, overexpression of Cks1 and Cks2 in esophageal carcinoma was closely associated with poor pathological features of esophageal carcinoma, including higher histologic grade of tumor, regional lymph nodes invasion, and neoplastic embolus. Overexpression of Cks1 and Cks2 is associated with the aggressive tumor behaviors of esophageal carcinoma. Further efforts are needed to determine whether overexpression of Cks1 and Cks2 can serve as novel biomarkers for esophageal carcinoma.

[The development of computerized cardiac programmable stimulator].

This paper introduces a fully computerized cardiac programmable stimulator. With the visual interface of computer, the configuration and control of stimulation will be realized conveniently. Meanwhile, the current status and the waves during stimulation will be displayed in window real-time, which improved the visualization and availability in medical application. It has been proved that this system is simpler in operation and more functional than traditional cardiac programmable stimulators, and therefore it is valuable in medical applications.

[The development of a 64-channel epicardial potential mapping system].

A 64-channel epicardial potential mapping system was developed in order to study the mechanics of arrhythmia such as atrial fibrillation and ventricular fibrillation, and instruct the procedure of detecting and eliminating abnormal rhythm in experiment or clinic. The system was consist of electrode, amplifier, A/D card, computer and output device. The system's software include signal acquisition module, signal preprocessing module, character-point detecting module, isochrone-map construction module, output display and print module. The system could be used to detect the activation path of atrial fibrillation and ventricular fibrillation.

[Ventricular wave identification in epicardial mapping system using wavelet transform].

A novel method for the identification of ventricular wave based on wavelet transform is presented. First, "Multi-Scale Intersectional Search" is adopted for analysis of wavelet transform coefficients on some different scales, then adaptive detection is used on this basis. The results of practice show that it has a discrimination of 94.5%, and is obviously superior to any routime method.

Prevalence of hepatitis C virus infection in Nanjing, southern China.

There is little information on the prevalence of hepatitis C virus (HCV) infection in China. The prevalence of HCV infection was determined in 998 subjects (398 with liver disease and 600 without) in the city of Nanjing in southern China. Subjects were tested for anti-HCV antibodies by a second generation assay. We also determined serological HCV genotypes and HCV RNA sequences. Among the 600 subjects without liver disease, 3 (0.5%) were seroreactive for anti-HCV. All were less than 12 years of age and had a history of transfusion. Of the 398 liver disease patients, 20 (5.0%) were seroreactive for anti-HCV. Of 16 patients in whom serological HCV genotype was determined, 10 (62.5%) were infected with HCV type 1, 5 (31.3%) with type 2 and 1 (6.3%) undetermined. HCV genomes sequenced from 2 patients belonged to genotype 2 and were closely related to strains in Beijing and Japan by molecular evolutionary analysis. These results suggest that HCV infection is rare and not a major cause of liver disease in southern China.

Factors affecting production of compound A from the interaction of sevoflurane with Baralyme and soda lime.

Various alkali (e.g., soda lime) convert sevoflurane to CF2=C(CF3)OCH2F, a vinyl ether called "Compound A, " whose toxicity raises concerns regarding the safe administration of sevoflurane via rebreathing circuits. In the present investigation, we measured the sevoflurane degradation and output of Compound A caused by standard (13% water) Baralyme brand absorbent and standard (15% water) soda lime, and Baralyme and soda lime having various water contents (including no water). We used a flow-through system, applying a gas flow rate relative to absorbent volume that roughly equaled the rate/volume found in clinical practice. Both absorbents, at similar water contents, temperatures, and sevoflurane concentrations, produced roughly equal concentrations of Compound A. Dry and nearly dry absorbents produced less Compound A early in exposure to sevoflurane, and more later, than standard absorbents. Increases in temperature and sevoflurane concentration increased output of Compound A. Both absorbents, especially when dry, also destroyed Compound A, the concentration exiting from absorbent resulting from a complex sum of production and destruction. We conclude that the variability of concentrations of Compound A found in clinical practice may be largely explained by the inflow rate used (i.e., by rebreathing), sevoflurane concentration, and absorbent temperature and dryness. The effect of dryness is complex, with fresh dry absorbent destroying Compound A as it is made, and with dry absorbent that has been exposed to sevoflurane for a period of time providing a sometimes unusually high output of Compound A.

Factors affecting the concentration of compound A resulting from the degradation of sevoflurane by soda lime and Baralyme in a standard anesthetic circuit.

Carbon dioxide absorbents, such as soda lime and Baralyme brand absorbent, convert sevoflurane to CF2 = C(CF3)OCH2F, a vinyl ether called "Compound A," whose toxicity raises concerns regarding the safety of sevoflurane in rebreathing circuits. Because an increased inflow rate to an anesthetic circuit decreases rebreathing, we assumed that an increased rate would proportionately decrease the concentration of Compound A. In the present report, we measured the Compound A concentration resulting from the action of wet (standard) soda lime and wet (standard) Baralyme on 2% sevoflurane in a model anesthetic circuit, using inflow rates (0.5, 1.0, 2.0, 4.0, and 6.0 L/min), ventilations (5 and 10 L/min), and carbon dioxide production/removal (200 and 400 mL/min) found in clinical practice. An increase in inflow rate decreased Compound A concentration to lower levels as inflow rate approached minute ventilation. At lower inflow rates, increasing duration of sevoflurane inflow increased the concentration of Compound A, a finding consistent with a progressive increase in absorbent temperature from absorption of carbon dioxide and consequently greater sevoflurane degradation. There was no material difference between Baralyme and soda lime in the concentrations of Compound A produced at a particular inflow rate. An increase in ventilation increased the concentration of Compound A, having a much greater effect at high rather than low inflow rates. An increase in amount of carbon dioxide absorbed also increased the concentration of Compound A. We conclude that inflow rate, ventilation, and carbon dioxide production are major determinants of the concentration of Compound A.

Carbon monoxide production from degradation of desflurane, enflurane, isoflurane, halothane, and sevoflurane by soda lime and Baralyme.

Anecdotal reports suggest that soda lime and Baralyme brand absorbent can degrade inhaled anesthetics to carbon monoxide (CO). We examined the factors that govern CO production and found that these include: 1) The anesthetic used: for a given minimum alveolar anesthetic concentration (MAC)-multiple, the magnitude of CO production (greatest to least) is desflurane > or = enflurane > isoflurane >> halothane = sevoflurane. 2) The absorbent dryness: completely dry soda lime produces much more CO than absorbent with just 1.4% water content, and soda lime containing 4.8% or more water (standard soda lime contains 15% water) generates no CO. In contrast, both completely dry Baralyme and Baralyme with 1.6% water produce high concentrations of CO, and Baralyme containing 4.7% water produces concentrations equaling those produced by soda lime containing 1.4% water. Baralyme containing 9.7% or more water and standard Baralyme (13% water) do not generate CO.3) The type of absorbent: at a given water content, Baralyme produces more CO than does soda lime. 4) The temperature: an increased temperature increases CO production. 5) The anesthetic concentration: more CO is produced from higher anesthetic concentrations. These results suggest that CO generation can be avoided for all anesthetics by using soda lime with 4.8% (or more) water or Baralyme with 9.7% (or more) water, and by using inflow rates of less than 2-3 L/min. Such inflow rates are low enough to ensure that the absorbent does not dry out.

Seroprevalence of hepatitis C virus infection and its genotype in Lanzhou, western China.

The seroprevalence of hepatitis C virus (HCV) infection in Lanzhou, Western China was studied. HCV genotypes in 20 patients with HCV infection was determined by genotype-specific primer for polymerase chain reaction (PCR) based on HCV core region and compared with the genotype assigned by sequence comparison and molecular evolutionary analysis based on the same region. Antibody to HCV (anti-HCV) was present in 2.5% of volunteer blood donors and in 35.0% of paid blood donors (P < 0.01). HCV infection is uncommon in patients with liver disease who attended liver clinics in this locality; 4.0% with acute hepatitis and 4.0% with chronic hepatitis, 10.0% with liver cirrhosis, and none with hepatocellular carcinoma were seropositive for anti-HCV. Genotype 1b and 2a were both found to be prevalent. Together, they accounted for 19 of 20 (95%) patients with HCV infection. Sequencing of the HCV core region from two patients showed that the assignment of HCV genotype by genotype-specific primers for PCR matched well with the genotyping results based on sequence comparison and molecular evolutionary analysis. These data showed that HCV is present in Western China, HCV infection is more common in paid blood donors, and HCV genotypes 1b and 2a are both prevalent in Western China.

Oxygen radicals in bronchoconstriction of guinea pigs elicited by isocapnic hyperpnea.

The role of oxygen radicals in isocapnic hyperpnea-induced bronchoconstriction (HIB) of guinea pigs was investigated using scavengers of the radicals. In series 1, 50 young guinea pigs were randomly divided into seven groups: control 1, control 2, chlorisondamine, tetrodotoxin (TTX), acute dimethylthiourea (DMTU), tachykinin depletion, and 5% CO2 in air. Animals of the control 2 group received vehicle (saline) infusion while those of the control 1 group did not. Chlorisondamine was used to block ganglionic transmission, TTX to interrupt nerve conduction, DMTU to scavenge hydroxyl radicals, and chronic capsaicin pretreatment to deplete tachykinins. The animals in the last group were ventilated with dry 5% CO2 in air during hyperpnea. In series 2, 13 additional animals were used to test the effects of intratracheal administration of superoxide dismutase and catalase (SOD + CAT) on HIB. Each animal was anesthetized with pentobarbital sodium, cannulated with a tracheal cannula and venous catheter, paralyzed with gallamine triethiodide, and mechanically ventilated. During the baseline period, each animal was ventilated normally with humidified air. Then it was hyperventilated 15 min with a dry gas mixture of 95% O2-5% CO2, except animals in the last group of series 1. Subsequently, all animals returned to normal ventilation with humidified air for 45 min (recovery period). The maximal expiratory flow and dynamic compliance were obtained periodically during the recovery period. The isocapnic hyperpnea using 95% O2-5% CO2, but not 5% CO2 in air, caused bronchoconstriction that was significantly blocked by acute DMTU, acute SOD + CAT, and tachykinin depletion. In an additional group of six animals, acute DMTU did not significantly alter acetylcholine-induced airway constriction.(ABSTRACT TRUNCATED AT 250 WORDS)

Noncholinergic airway constriction: role of axon reflex and oxygen radicals.

In the airways and lungs, activated afferent C-fibers release tachykinins, which induce noncholinergic bronchoconstriction. We have, for several years, focused our research on the role of axon reflex and oxygen radicals in noncholinergic airway constriction in guinea pigs. In this species, the noncholinergic bronchial constriction is not affected by administration of a ganglionic blocking agent, chlorisondamine, indicating that only the afferent C-fiber, not a complete reflex arc, is required for this constriction. Accordingly, we investigated the role of axon reflex by using tetrodotoxin (TTX) and bupivacaine to block impulse conduction in the axon. For the role of oxygen radicals, superoxide dismutase (SOD), catalase (CAT), and dimethylthiourea (DMTU) were employed to scavenge superoxide anion, hydrogen peroxide, and hydroxyl radical, respectively. We used capsaicin, resiniferatoxin (RTX), or hyperventilation to activate afferent C-fibers which, in turn, release tachykinins and lead to noncholinergic airway constriction. The constriction was quantified by measuring the maximal expiratory flow rate (Vmax) and dynamic compliance (Crs). Both capsaicin and RTX caused an immediate decrease in Vmax and Crs, indicating severe bronchoconstriction. This constriction decreased gradually with time. Tachykinin depletion abolished neurotoxin-induced airway constriction, suggesting that tachykinins mediate the constriction. Both TTX and bupivacaine significantly attenuated the constriction at 15-20 min after neurotoxin administration. Therefore, these data suggest that the axon reflex plays an important role in noncholinergic bronchial constriction. In other studies, capsaicin- or hyperventilation-induced bronchoconstriction was abolished by tachykinin depletion, as well as significantly ameliorated by the administration of antioxidants. These data indicate that oxygen radicals modulate the tachykinin-mediated noncholinergic airway constriction.

Inhibition of [3H]dopamine uptake into rat striatal slices by quaternary N-methylated nicotine metabolites.

The effects of quaternary N-methylated nicotine derivatives were examined on in vitro uptake of [3H]dopamine ([3H]DA) in rat striatal slices. Striatal slices were incubated with a 10 microM concentration of the following compounds: N-methylnicotinium, N-methylnornicotinium, N-methylcotininium, N,N'-dimethylnicotinium and N'-methylnicotinium salts. The results clearly indicated that significant (60%) inhibition of [3H]DA uptake occurred with those compounds possessing a N-methylpyridinium group; whereas, compounds that were methylated at the N'-pyrrolidinium position were less effective or exhibited no inhibition of [3H]DA uptake. The results suggest that high concentrations of quaternary N-methylated nicotine metabolites which are structurally related to the neurotoxin MPP+, and which may be formed in the CNS, may protect against Parkinson's Disease and explain the inverse relationship between smoking and Parkinsonism reported in epidemiologic studies.

[Use of signal averaging system in the study of ventricular late potentials].

Signal-averaged electrocardiograms (SAECGs) were performed in 93 patients with various heart diseases and 20 dogs with experimental acute myocardial infarction. SA-ECGs were recorded with a high-pass bidirectional digital filter Mode FDS which was made by Shanghai Fudan University and Suzhou Medical College. All the patients and dogs were also examined with ART 1200 EPX recorder. Ventricular late potentials (VLPs) were directly recorded from the epicardium with experimental infarction by using composite electrodes and signal averaging system in late stage of coronary ligation. The results showed: VLPs were detected by body surface SA-ECG only in half of the dogs with VLPs detected by the real time mapping on infarction region, but the abnormal SAECGs did reveal the delayed ventricular activity; the data obtained with FDS or ART SA-ECG were similar.

On the structure selectivity problem in drug design. A comparative study of benzylpyrimidine inhibition of vertebrate and bacterial dihydrofolate reductase via molecular graphics and quantitative structure-activity relationships.

Quantitative structure-activity relationships (QSAR) have been derived for the action of 68 5-(substituted benzyl)-2,4-diaminopyrimidines on dihydrofolate reductase (DHFR) from Lactobacillus casei and chicken liver. The QSAR are analyzed with respect to the stereographics models of the active sites of the enzymes and found to be in good agreement. Using these QSAR equations, we have attempted to design new trimethoprim-type antifolates having higher selectivity for the bacterial enzyme. The general problem of developing selective inhibitors is discussed.