[Questionnaire survey analysis on the screening of tuberculosis among diabetic patients in general hospitals of Hunan Province].

Objective: To investigate the development of tuberculosis screening-related tests in general public hospitals(GPHs) of different levels in Hunan Province and the"awareness and practice of screening tuberculosis in diabetic patients"by doctors directly involved in diabetes diagnosis and treatment in the hospitals, aiming to provide reference for the formulation of the tuberculosis-diabetes joint prevention and control activity plan based on our national conditions. Methods: Stratified sampling was used to select 43 GPHs at three different levels in Hunan province: 14 tertiary GPHs, 13 secondary GPHs, and 16 primary GPHs. 284 endocrinologists working in enrolled hospitals were invited to participate in the on-site questionnaire-survey and 277 qualified. The study used SPSS 22.0 statistical software to analyze the data. The prevalence rate of tuberculosis screening test among hospitals at all levels was compared by chi-square test, and logistic regression was used to analyze the related factors affecting doctors' screening awareness. P≤0.05 was considered statistically significant. Results: The allocation of digital X-ray cameras, tuberculin skin tests, sputum acid-fast bacillus smears, sputum cultures for Mycobacterium tuberculosis, and interferon-gamma release assays in the 43 GPHs were 90.7% (39/43), 72.1% (31/43), 55.8% (24/43), 34.9% (15/43), 27.9% (12/43) with significant differences between the different hospital levels(P<0.05). 173 endocrinologists considered it necessary to proactively initiate tuberculosis screening for patients at first diagnosis. When admitting patients, 197 endocrinologists chose tuberculosis screening only for diabetes mellitus patients with suspected tuberculosis symptoms. The most possible reasons why diabetes mellitus patients wouldn't undergo tuberculosis screening were"patients refused(76.5%, 212/277)","patients didn't complain of the symptom(46.9%, 130/277)", and"tuberculosis screening-related tests haven't been conducted in the hospital(35.7%, 99/277)". Conclusions: Although endocrinologists displayed some tuberculosis-related knowledge and awareness of the need for proactive tuberculosis screening, the actual screening rate in the clinical setting was low. This may be related to multiple factors, including those of patients, doctors, and medical institutions.

[Review on risk factors associated with tuberculosis death].

Tuberculosis is a major public health problem threatening human health and life. Although the level of diagnosis and treatment of tuberculosis has greatly improved in recent years, it is still one of the major infectious diseases causing human death. Tuberculosis has the characteristics of both chronic and infectious diseases, and is affected by many factors. Identification of the risk factors associated with tuberculosis death has certain guiding value for the prevention and control of tuberculosis. This articled reviewed recent studies on risk factors associated with tuberculosis death both in China and abroad. Male, old age, smoking, drinking, low socioeconomic level, malnutrition, drug resistance, AIDS and diabetes mellitus were found to increase the risk of tuberculosis mortality. It is expected that these findings could provide useful information to clinicians for taking necessary interventions to reduce tuberculosis mortality.

[Effect of smoking on chronic rhinosinusitis complicated with lower respiratory tract disease].

Objective: To study the effect of smoking on chronic rhinosinusitis with lower respiratory tract diseases and underlaying mechanism. Method: The data of 54 patients diagnosed as CRS with lower respiratory disease (Group A) and 60 patients with CRS (Group B) from otolaryngology head and neck surgery in Wuhan Central Hospital and Renmin Hospital of Wuhan University from January 2014 to December 2017 were analyzed, retrospectively. According to the history of smoking, all patients were divided into smoking group (Group C) and non-smoking group (Group D). Nasal secretory microbial cultures were performed. Result: Patients with CRS with lower respiratory disease had higher smoking rate than patients with CRS alone(P<0.05). CRS patients with smoking increased the rate of bacterial infection in lower respiratory and increased the incidence of lower respiratory disease in patients with CRS (P<0.05). Conclusion: Smoking is an important risk factor for CRS complicated with lower respiratory disease.

Molecular cloning and functional characterization of cyclin E and CDK2 from Penaeus monodon.

Reduced reproductive performance of the black tiger shrimp (Penaeus monodon) has caused economic losses and hampered the fishing industry. Detailed investigation of the molecular mechanism by which the cell cycle is regulated in this organism is needed to understand the development and maturation of ovaries and oocytes, with a view to improving reproductive capacity. Cell cycle progression is mainly determined by cyclin-dependent kinase (CDK) and cyclin complexes, the cyclin E/CDK2 complex playing a key role in G1/S transition. However, knowledge of the interplay between cyclin E and CDK2 in invertebrates remains limited. In this study, full-length P. monodon cyclin E (Pmcyclin E) and CDK2 (PmCDK2) sequences were cloned. The open reading frame of Pmcyclin E was 1263 bp in length and encoded a 47.9-kDa protein, while that of PmCDK2 was 921 bp, encoding a protein of 34.9 kDa. Recombinant cyclin E and CDK2 proteins were expressed in Escherichia coli and purified by Ni-chelating affinity chromatography. In addition, a pull-down assay was performed to identify any interaction between Pmcyclin E and PmCDK2. This research provides a basis for the study of the functional mechanisms of the cyclin E/CDK2 complex in shrimp, further enriching our knowledge of invertebrate cell cycle regulation.

Molecular cloning and expression analysis of MAT1 gene in black tiger shrimp (Penaeus monodon).

MAT1 (ménage à trois 1), an assembly factor and targeting subunit of the CDK-dependent kinase (CAK), can regulate the cell cycle, transcription, and DNA repair. This study was intended to investigate the role of MAT1 in the reproductive maturation of black tiger shrimp (Penaeus monodon). In this study, the P. monodon MAT1 (PmMAT1) gene was identified and characterized. The full-length cDNA of PmMAT1 was 1490 bp in length with an open-reading frame of 993 bp corresponding to 330 amino acids. The temporal expression of PmMAT1 in various tissues was measured by quantitative real-time PCR with the highest expression observed in ovaries. In the ovaries, the PmMAT1 gene was continuously but differentially expressed during the maturation stages. Comparative analyses of MAT1, CDK7, and cyclin H in the CAK complex of P. monodon indicated that the expression of CDK7 and cyclin H coincided with that of MAT1 during the ovary maturation stages. Serotonin (5-HT) injection promoted the expression level of PmMAT1 in the ovaries of shrimp at 6-48 h post-injection. These results indicate that PmMat1 plays a prominent role in the process of ovarian maturation.

Characterization and expression analysis of a cyclin B gene from black tiger shrimp (Penaeus monodon).

The open reading frame of black tiger shrimp (Penaeus monodon) cyclin B (Pmcyclin B) was identified, based on cDNA sequence registered in GenBank (accession No. EF015590). The target sequence was 1206 bp, corresponding to 401 amino acids. Two conserved signature sequences of the cyclin B gene family were found in the Pmcyclin B deduced aa sequence. Temporal expression of Pmcyclin B in different tissues, including ovary, lymphoid organ, brain, blood, muscle, heart, gill, hepatopancreas, and intestine, were quantified by quantitative real time PCR. Messenger RNA expression levels of Pmcyclin B were greatest in the ovary, compared to other tissues (P < 0.05). Temporal expression of Pmcyclin B in the ovary at six different developmental stages was investigated by real-time PCR; no significant difference was observed (P < 0.05). Recombinant Pmcyclin B protein and its polyclonal antibody were successfully produced. Western blot analysis revealed differential expression of Pmcyclin B in ovaries in developmental stages II to IV; a positive signal (45 kDa) was observed in all ovarian stages assessed, but was most intense at stage III. Pmcyclin B protein was assessed by immunohistochemistry and was localized to the cytoplasm of prophase oocytes at stage II and enriched in the nuclei of pro-metaphase oocytes at stages III and IV. Results from this study indicate that Pmcyclin B is constitutively expressed and plays an important role in ovarian maturation in P. monodon.

Involvement of cerulospinal glutamatergic neurotransmission in fentanyl-induced muscular rigidity in the rat.

BACKGROUND: Investigators in the authors' laboratory previously established the critical participation of the cerulospinal noradrenergic pathway in muscular rigidity elicited by fentanyl. The identification of colocalization of glutamate with tyrosine hydroxylase in most locus ceruleus neurons suggests a role for cerulospinal glutamatergic neurotransmission in fentanyl-induced muscular rigidity. This suggestion and the subtype(s) of glutamate receptors involved were investigated here. METHODS: Electromyographic signals activated by bilateral microinjection of 2.5 microg fentanyl into the locus ceruleus were recorded differentially from the left sacrococcygeus dorsi lateralis muscle of adult male Sprague-Dawley rats. The effect of intrathecal administration at the lower lumbar spinal cord of various N-methyl-D-aspartate (NMDA) and non-NMDA receptor antagonists or agonists on this index of muscular rigidity was studied. Rats were under mechanical ventilation, and intravenous infusion of ketamine (30 mg x kg(-1) x h(-1)) was maintained until 10 min before fentanyl was administered. RESULTS: Microinjection of fentanyl bilaterally into the locus ceruleus increased the root mean square and decreased the mean power frequency values of electromyographic signals. The efficacy of fentanyl to elicit muscular rigidity in this manner was significantly reduced by previous intrathecal administration of either 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate (MK-801), D-(-)-2-amino-5-phosphonovaleric acid (AP5), or (+/-)-3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP). Intrathecal administration of kainic acid or NMDA also resulted in significant electromyographic activation. CONCLUSIONS: In addition to the cerulospinal noradrenergic mechanism, the cerulospinal glutamatergic pathway and both NMDA and non-NMDA receptors in the spinal cord may mediate fentanyl-induced muscular rigidity in the rat.

Inhibition by intrathecal prazosin but not yohimbine of fentanyl-induced muscular rigidity in the rat.

We evaluated the effect of intrathecally administered prazosin, alpha 1-adrenoceptor antagonist, or yohimbine, alpha 2-adrenoceptor antagonist, on fentanyl-induced muscular rigidity. Adult, male Sprague-Dawley rats were anesthetized with ketamine and were under mechanical ventilation. Fentanyl given intravenously (100 micrograms/kg) or microinjected into the bilateral locus coeruleus (LC) (2.5 microgram/50 nl) consistently evoked a significant increase in the electromyographic activity recorded from the sacrococcygeus dorsalis lateralis muscle. This implied muscular rigidity was appreciably antagonized by prior intrathecal (10 microliters) administration of prazosin (5 or 10 nmol), but not equimolar dose of yohimbine. These results suggest that the spinal alpha 1-adrenoceptors in the coerulospinal noradrenergic pathway play a key role in fentanyl-induced muscular rigidity.

Involvement of potassium and calcium channels at the locus coeruleus in fentanyl-induced muscular rigidity in the rat.

Previous work from our laboratory suggested that Go alpha protein at the locus coeruleus (LC) may be involved in the signal transduction process that underlies muscular rigidity induced by fentanyl. The present study further evaluated the roles of K+ and L-type Ca2+ channels, gating of which is known to be associated with activation of Go alpha protein, in this process, using Sprague-Dawley rats anesthetized with ketamine. Bilateral microinjection into the LC of tetraethylammonium chloride (100 or 200 pmol), a K+ channel blocker, and S(-)-Bay K 8644 (0.5 nmol), a Ca2+ channel activator, produced significant antagonization of the EMG activation elicited by fentanyl (100 micrograms/kg, i.v.), as recorded from the sacrococcygeus dorsalis lateralis muscle. On the other hand, local application to the bilateral LC of diazoxide (10 or 20 nmol), an ATP-dependent K+ channel activator, and nifedipine (0.25 or 0.5 pmol), a L-type Ca2+ channel blocker, was ineffective in blunting fentanyl-induced muscular rigidity. These results suggest that activation of K+ channels and/or inhibition of L-type Ca2+ channels secondary to triggering of the Go alpha protein at the LC may underlie the signal transduction process in the mediation of fentanyl-induced muscular rigidity.

Subtypes of Guanine-Nucleotide-Binding Regulatory Proteins at the Locus coeruleus Involved in Fentanyl-Induced Muscular Rigidity in the Rat.

Previous results from our laboratory have established that the G(o) subtype of guanine nucleotide (GTP)-binding regulatory protein at the locus coeruleus (LC) may participate in the elicitation of muscular rigidity by fentanyl. The present study further examined the involvement of other subtypes of GTP-binding regulatory proteins at the LC in this process, using Sprague-Dawley rats anesthetized with ketamine (120 mg/kg, i.p., with 30 mg/kg/h i.v. infusion supplements) and under mechanical ventilation. Intravenous administration of fentanyl (100 &mgr;g/kg) induced a significant increase in electromyographic signals recorded from the sacrococcygeus dorsi lateralis muscle. Power spectral analysis revealed that this was accomplished by a decrease in the mean power frequency and an increase in the root mean square values of the signals. The above responses were appreciably antagonized by pretreating animals with bilateral microinjection into the LC of pertussis toxin (80 or 160 fmol), N-ethylmaleimide (16 pmol) or 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (100 or 200 fmol); but not by cholera toxin (120 or 240 fmol), forskolin (240 or 480 pmol) or N-ethylmaleimide at a higher dose (32 pmol). These results suggest that, in addition to G(o) protein, fentanyl-induced muscular rigidity may also involve other pertussis toxin-sensitive GTP-binding regulatory proteins, possibly G(i) and G(p) subtypes, in the signal transduction processes following activation of &mgr;-opioid receptors at the LC. Copyright 1995 S. Karger AG, Basel

Aluminum-induced alterations in [3H]ouabain binding and ATP hydrolysis catalyzed by the rat brain synaptosomal (Na(+)+K+)-ATPase.

The (Na(+)+K+)-ATPase is responsible for maintenance of the ionic milieu of cells. The objective of this study is to investigate the effect of aluminum, an ion implicated in several neurological disorders, on ATP hydrolysis catalyzed by the rat brain synaptosomal (Na(+)+K+)-ATPase and on the binding of [3H]ouabain to this enzyme. AlCl3 (25-100 microM) inhibits the phosphatase activity of the (Na(+)+K+)-ATPase in a dose-dependent manner. AlCl3 appears to act as a reversible, noncompetitive inhibitor of (Na(+)+K+)-ATPase activity by decreasing the maximum velocity of the enzyme without significantly affecting the apparent dissociation constant with respect to ATP. AlCl3 may affect Mg2+ sites on the (Na(+)+K+)-ATPase but does not appear to interact with Na+ or K+ sites on the enzyme. In contrast to this inhibitory effect on the phosphatase function of the enzyme, AlCl3 (1-100 microM) stimulates the binding of [3H]ouabain to the (Na(+)+K+)-ATPase. This effect is due to an increase in the maximum [3H]ouabain binding capacity of the enzyme with no change in the [3H]ouabain binding affinity. These data support the hypothesis that AlCl3 may stabilize the phosphorylated form of the synaptosomal (Na(+)+K+)-ATPase which increases [3H]ouabain binding while inhibiting the phosphatase activity of the enzyme.

Power spectral analysis of electromyographic and systemic arterial pressure signals during fentanyl-induced muscular rigidity in the rat.

We have measured electromyographic (EMG) and systemic arterial pressure (SAP) signals during fentanyl-induced muscular rigidity in adult male Sprague-Dawley rats anaesthetized initially with ketamine 120 mg kg-1 i.p. during controlled ventilation. Fentanyl 100 micrograms kg-1 i.v. induced significant increase in EMG activity, recorded from the sacrococcygeus dorsi lateralis muscle. Power spectral analysis revealed that this was produced by an increase in the root mean square and a decrease in the mean power frequency values of the signals, signifying recruitment and synchronous activation of motor units. Together with transient hypotension and bradycardia, power spectral analysis of the SAP signals demonstrated a reduced but maintained power density of the frequency components that represent respiratory, baroreceptor and vasomotor activities. All these effects were only demonstrated unequivocally in rats maintained by i.v. infusion of ketamine until 10 min before the administration of fentanyl. We conclude that analysis of the temporal alterations in the spectral components of the EMG and SAP signals in rats during mechanical ventilation provides a sensitive method of measuring fentanyl-induced muscular rigidity and the accompanying alterations in haemodynamic variables.

Involvement of G(o) alpha subtype of guanine nucleotide-binding regulatory protein at the locus coeruleus in fentanyl-induced muscular rigidity in the rat.

Previous work from our laboratory suggested that locus coeruleus (LC) and the coerulospinal noradrenergic pathway are intimately related to the elicitation of muscular rigidity by fentanyl. The present study attempted to identify the subtype of guanine nucleotide-binding regulatory protein that may participate in this process, using Sprague-Dawley rats anesthetized with ketamine and under mechanical ventilation. Immunofluorescent staining with a polyclonal antiserum directed against a 39-kDa protein that corresponds to the alpha subunit of G(o) revealed the presence of G(o) alpha immunoreactivity in neurons of the LC. Bilateral microinjection of the same G(o) alpha antiserum into the LC also significantly blunted the enhanced electromyographic activity recorded from the sacrococcygeus dorsalis lateralis muscle induced by intravenous administration of fentanyl (100 micrograms/kg). These results suggest that G(o) alpha protein at the LC may participate in the signal transduction process that underlies muscular rigidity induced by high-dose fentanyl.

Control of the Na+,K(+)-ATPase under normal and pathological conditions.

The Na+,K(+)-ATPase is an important enzyme in determining the ionic milieu of the cerebromicrovasculature and neurons. The effect of hypertension or aging on this enzyme, as well as its susceptibility to regulation by fatty acids or aluminum, is the focus of this study. A significant increase (34%) in the apparent affinity constant (KD) but no change in the maximum binding capacity (Bmax) for [3H]ouabain binding to the cerebromicrovascular Na+,K(+)-ATPase occurs after induction of acute hypertension. In addition, long chain unsaturated fatty acids stimulate the binding of [3H]ouabain to the enzyme in microvessels from normotensive and hypertensive rats. The synaptosomal Na+,K(+)-ATPase is sensitive to aluminum. AlCl3 (1-100 microM) inhibits the K(+)-dependent-p-nitrophenylphosphatase (K(+)-NPPase) activity of the Na+,K(+)-ATPase in a dose-dependent manner. AlCl3 (100 microM) decreases the Vmax by 14% but does not alter the KM, suggestive of non-competitive inhibition. The enzyme from aged brain displays a greater Vmax, but shows the same susceptibility to AlCl3 as the enzyme from younger brain. In summary, disruption of the Na+,K(+)-ATPase may underlie, at least in part, abnormalities of nerve and vascular cell function in disorders where elevated concentrations of fatty acids or metal ions are involved.

Participation of pertussis toxin-sensitive GTP-binding regulatory proteins in the suppression of baroreceptor reflex by neurotensin in the rat.

We evaluated the molecular mechanism that may underlie the suppressive effect of neurotensin (NT) on the baroreceptor reflex (BRR), using Sprague-Dawley rats that were anesthetized with sodium pentobarbital (50 mg/kg, i.p.). Intracerebroventricular (i.c.v.) application of NT (15 nmol) significantly inhibited the BRR response. Such an inhibition was appreciably antagonized by pretreating animals with i.c.v. injection of pertussis toxin (10 or 20 pmol), N-ethylmaleimide (1 or 2 nmol), forskolin (30 or 60 nmol) or phorbol 12-myristate 13-acetate (2 or 4 nmol), but not by cholera toxin (15 or 30 pmol). More specifically, pretreatments with bilateral microinjection into the nucleus tractus solitarius (NTS) of pertussis toxin (80 or 160 fmol), N-ethylmaleimide (80 pmol), forskolin (480 pmol) or phorbol 12-myristate 13-acetate (16 or 32 pmol) also blunted the NT-induced suppression of BRR, although cholera toxin (120 or 240 fmol), or 1,9-dideoxyforskolin (480 pmol) had no appreciable effect. These results suggest that a pertussis toxin-sensitive guanine nucleotide-binding regulatory protein(s), which is not likely to be Gs, possibly Gi or Gp, may be involved in the transmembrane signaling process that underlies the suppression of BRR response by NT at the NTS.