[Species and population density of malaria vector Anopheles in Sichuan Province from 2011 to 2021].

OBJECTIVE: To investigate the distribution of malaria vector Anopheles in Sichuan Province from 2011 to 2021, so as to provide the scientific evidence for improving the surveillance of malaria vector Anopheles and preventing re-establishment of imported malaria in Sichuan Province. METHODS: The density and species of Anopheles mosquitoes were investigated using human-bait trapping and light trapping techniques in malaria vector surveillance sites of Sichuan Province from 2011 to 2021. The number, population and density of captured Anopheles mosquitoes were collected and descriptively analyzed, and the geographical distribution map of malaria vectors was plotted using the software ArcGIS 10.7 in Sichuan Province. RESULTS: A total of 152 243 Anopheles mosquitoes were captured in malaria vector surveillance sites of Sichuan Province from 2011 to 2021, including 150 987 An. sinensis (99.18%) and 1 256 An. anthropophagus (0.82%), and no other Anopheles species were captured. The annual densities of An. sinensis and An. anthropophagus were 0.64 to 1.27 mosquitoes/(person-hour) and 0 to 0.07 mosquitoes/(person-hour) by the human-bait trapping technique, and 6.46 to 26.50 mosquitoes/(light-night) and 0 to 0.82 mosquitoes/(light-night) by the light trapping technique in malaria vector surveillance sites of Sichuan Province from 2011 to 2021. A relatively higher density of An. anthropophagus was seen in Renshou County, Jianyang City, Weiyuan County and Mabian Yi Autonomous County [> 0.40 mosquitoes/(person-hour)] by the human-bait trapping technique, and in Cuiping District and Gaoxian County in Yibin City [> 1.00 mosquito/(light-night)] by the light trapping technique in Sichuan Province from 2011 to 2018, with no An. anthropophagus captured from 2019 to 2021, and a relatively higher density of An. sinensis was detected in Emeishan City, Lushan County, Luojiang District, Tongchuan District and Zhaohua District [> 4.00 mosquitoes/(person-hour)] by the human-bait trapping technique, and in Huili County, Yuexi County, Dechang County, Langzhong City, Pingchang County and Xuanhan County [> 40.00 mosquitoes/(light-night)] by the light trapping technique in Sichuan Province from 2011 to 2021. CONCLUSIONS: Malaria vectors were still widespread in Sichuan Province from 2011 to 2021, and An. sinensis was the dominant species of malaria vectors. There is still a risk of local re-establishment of imported malaria in Sichuan Province, and it is needed to continue to improve the surveillance of imported malaria cases and malaria vectors.

[Analysis of laboratory re-examination results in Sichuan Provincial Malaria Diagnostic Reference Laboratory from 2014 to 2020].

OBJECTIVE: To re-examine the diagnosis results of reported malaria cases in Sichuan Province from 2014 to 2020, so as to assess the malaria diagnostic capability of Sichuan Provincial Malaria Diagnostic Reference Laboratory. METHODS: The blood and blood smear samples from reported malaria cases were collected by Sichuan Provincial Malaria Diagnostic Reference Laboratory from 2014 to 2020, and subjected to re-examinations using microscopy and nested PCR assay. The re-examination results were compared. RESULTS: A total of 1 710 samples from reported malaria cases were re-examined by Sichuan Provincial Malaria Diagnostic Reference Laboratory from 2014 to 2020, and 1 634 samples were identified positive, with a positive coincidence rate of 95.56% (1 634/1 710) and a 92.29% (1 508/1 634) total coincidence rate of the Plasmodium species. The coincidence rates with P. falciparum, P. vivax, P. malariae and P. ovale were 99.48% (961/966), 97.07% (430/443), 83.05% (98/118) and 67.86% (19/28), respectively, and the coincidence rate was 91.81% (1 513/1 648) between microscopic and nested-PCR results. CONCLUSIONS: The capability of microscopists remains weak at grassroot medical institutions in Sichuan Province. Further training is required among microscopists to improve the malaria surveillance capability in Sichuan Province during the post-elimination stage.

Identification of Type H Vessels in Mice Mandibular Condyle.

Type H vessel is a specific vessel subtype that is strongly positive for CD31 and endomucin (CD31hiEmcnhi). It has already been identified that it can tightly regulate the coupling of angiogenesis and osteogenesis in the long bone of mice and human beings. The long bone is formed through endochondral ossification, which is the same type of process happening in mandibular condyle. Although the ossification of long bone and mandibular condyle has the same developmental process, the existence of type H vessels in the mouse condyle remains unclear. To address this, we identified that abundant type H vessels existed in the subchondral bone of the mouse condylar head and endosteum of the mouse condylar neck. Meanwhile, immunofluorescence imaging of the condyles in different ages of male C57BL/6J mice demonstrated that type H vessels decreased while aging. Furthermore, we validated a positive correlation between type H vessels and Osterix+ osteoprogenitors in the condyle induced by mandibular advancement. Mechanistically, we confirmed that deferoxamine mesylate, which promoted the proliferation of type H endothelial cells by activating hypoxia-inducible factor 1α (HIF-1α) signaling pathways, largely prevented the osteopenia in the condyle induced by botulinum toxin type A. Collectively, these results demonstrate that in the mouse condyle, type H vessels in areas of high function positively correlate with bone formation. In addition, we show a novel influence of HIF-1α signaling on osteogenesis via an increase in type H vessels. In conclusion, promoting angiogenesis of type H vessels is a promising strategy for the therapeutic improvement of osteogenesis in mandibular condyle.

Sdccag3 Promotes Implant Osseointegration during Experimental Hyperlipidemia.

Hyperlipidemia adversely affects bone metabolism, often resulting in compromised osseointegration and implant loss. In addition, genetic networks associated with osseointegration have been proposed. Serologically defined colon cancer antigen 3 (Sdccag3) is a novel endosomal protein that functions in actin cytoskeleton remodeling, protein trafficking and secretion, cytokinesis, and apoptosis, but its roles in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and in implant osseointegration under hyperlipidemic conditions have not been uncovered. Here, we performed microarray and RNA sequencing analysis to determine the differential expression of the Sdccag3 gene and related noncoding RNAs (ncRNAs) and to assess the long noncoding RNA (lncRNA) MSTRG.97162.4-miR-193a-3p-Sdccag3 coexpression network in bone tissues within the region 0.5 mm around implants in hyperlipidemic rats. In this experiment, we found that Sdccag3 and the previously uncharacterized lncRNA-MSTRG.97162.4 were downregulated during hyperlipidemia, while miR-193a-3p was upregulated. Sdccag3 overexpression increased new trabecular formation, the bone volume/total volume (BV/TV) (1.24-fold), and bone-implant combination ratio (BIC%) (1.26-fold). An RNA pulldown experiment revealed that Sdccag3 protein targeted lncRNA-MSTRG.97162.4 nucleotides 361 to 389. In addition, lncRNA-MSTRG.97162.4 overexpression significantly enhanced Sdccag3 (2.78-fold) expression and increased BV/TV (1.45-fold) and BIC% (1.07-fold) at the bone-implant interface. Taken together, these findings indicate that Sdccag3 overexpression enhances implant osseointegration under hyperlipidemic conditions by binding to lncRNA-MSTRG.97162.4. Furthermore, miR-193a-3p overexpression inhibited lncRNA-MSTRG.97162.4 (0.63-fold) and Sdccag3 (0.88-fold) expression and induced poor implant osseointegration (BV/TV, 0.86-fold; BIC%, 0.82-fold), while miR-193a-3p downregulation produced the opposite results (lncRNA-MSTRG.97162.4, 10.69-fold; Sdccag3, 6.96-fold; BV/TV, 1.20-fold; BIC%, 1.26-fold). Therefore, our findings show that Sdccag3 promotes implant osseointegration, and its related lncRNA-MSTRG.97162.4 and miR-193a-3p play an important role in osseointegration during hyperlipidemia, which might be a promising therapeutic target for improving dental implantation success rates.

[Outcome of surgical management and pathogenesis of female primary bladder neck obstruction].

OBJECTIVE: To investigate the effect of transurethral resection of bladder neck on primary female bladder neck obstruction and to analyze the expression of three kinds of sex hormone receptor (SR) in female bladder neck tissues diagnosed as primary bladder neck obstruction by the immunochemistry and statistics. METHODS: The clinical data of 40 female patients, admitted into Peking University People's Hospital for difficulty of voiding during Oct.2008 and Dec.2013 and eventually diagnosed as bladder outlet obstruction (BOO) by urodynamics, were retrospectively reviewed. BOO was defined as a maximum flow rate (Qmax) less than 12 mL/s together with a detrusor pressure at maximum flow rate (Pdet Qmax) more than 25 cmH2O in urodynamic study in the absence of neurological disorders. Diagnosis was confirmed by the cystoscopy. Preoperative and postoperative AUASS scores were recorded and analyzed for observation of curative effects and complications. The immunochemical expression of SR of primary female bladder neck obstruction (PBNO) tissues and normal control was examined and applied to statistical analysis. RESULTS: There were significant changes postoperatively in voiding scores, storage scores and total scores (P<0.001). Postoperatively, 1 patient newly presented with overactive bladder (OAB), 4 patients newly presented with hematuria, and 1 patient underwent cystostomy. The symptoms of urinary retention with overflow incontinence in 2 patients disappeared after the surgery, and 3 patients complicated with OAB complained without urgency. In addition, pre-hydronephrosis improved postoperatively in six patients. The subjective satisfactory rate to the surgery of TURBN was 77.5% (31/40). Sex hormone receptor, including androgen receptor (AR), estrogen receptor (ER), progesterone receptor (PR), expressed in both bladder neck tissues of normal control and PBNO patients. In PBNO group, the expression of PR was significantly lower than that of control group (P<0.05), while the other 2 SRs expressed with no significantly statistical difference. PBNO patients were divided into 2 groups, according to their symptoms scores, and the expression of SRs showed no significant differences among the mild, moderate and severe groups (P>0.05). CONCLUSION: The transurethral bladder neck resection is valid in treating with female PBNO patients, with rarely occurrence of complications. PR expressed less in the female bladder neck tissues, and is possibly correlated with the occurrence of female PBNO.

LncRNA LACAT1 promotes proliferation of oral squamous cell carcinoma cells by inhibiting microRNA-4301.

OBJECTIVE: To investigate the expression of lncRNA LACAT1 in oral squamous cell carcinoma (OSCC) tissues, and to further investigate its potential mechanism in OSCC as well as its relationship with clinicopathology and prognosis. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to analyze LACAT1 level in 78 pairs of OSCC tumor tissues and adjacent tissues, and the interplay between LACAT1 level and OSCC clinical indices along with patient's prognosis was analyzed. Further, the expression level of LACAT1 in OSCC cell line was verified by qRT-PCR. In addition, after LACAT1 knockdown model was constructed using lentivirus and transfected into OSCC cell lines, cell cloning assay and flow cytometry were used to analyze the impact of LACAT1 on biological functions of OSCC cells, and finally its association with microRNA-4301 was explored. RESULTS: In this experiment, qRT-PCR results revealed that LACAT1 level in OSCC tumor tissue specimens was significantly higher than that in paracancerous tissues. In addition, the pathology stage in patients with higher level of LACAT1 was also higher while the overall survival rate was lower. Compared with sh-NC group, the cell proliferation ability of sh-LACAT1 group was significantly decreased while cell apoptosis was oppositely increased. QRT-PCR results showed that microRNA-4301 level was significantly elevated in cells of sh-LACAT1 group, suggesting that the expression of above two molecules was negatively correlated. Meanwhile, cell reverse experiment also demonstrated that LACAT1 and microRNA-4301 can be mutually regulated, and thereby promote the malignant progression of OSCC. CONCLUSIONS: The LACAT1 level was found significantly increased in OSCC tissues and cells, which resulted in an advanced OSCC pathological stage and a poor prognosis of patients. In addition, it was found that LACAT1 may promote the malignant progression of OSCC by modulating microRNA-4301 activity.

[Antimicrobial susceptibility and genotyping of Mycobacterium intracellulare].

Objective: To investigate the antimicrobial susceptibility and genotyping of Mycobacterium intracellulare. Methods: A total of 150 M. intracellulare isolates were collected. The susceptibility against 15 antimicrobial agents widely used for treatment of non-tuberculosis mycobacteria (NTM) infections, was tested by broth microdilution assay. Variable number of tandem repeats (VNTR) assay was also performed using the 16-loci genotyping method. Results: The drug susceptibility test revealed that clarithromycin (97.3%, 146/150), moxifloxacin (94.0%, 141/150) and amikacin (90.0%, 135/150) had the best antimicrobial activities in vitro against the M. intracellulare isolates. Secondly, 75.3%(113/150), 64.0%(96/150), 52.7%(79/150) and 8.7%(13/150) of the strains were susceptible to rifampicin, linezolid, capreomycin, and ethambutol, respectively. The MIC(50) and MIC(90) values of the 3 injectable anti-tuberculosis drugs were as follows: amikacin 4 mg/L and 16 mg/L, streptomycin 4 mg/L and 16 mg/L, capreomycin 8 mg/L and 16 mg/L. The MIC(50) and MIC(90) values of the 5 different fluoroquinolones were 0.5 mg/L and 2 mg/L for moxifloxacin , 1 mg/L and 8 mg/L for ciprofloxacin, 1 mg/L and 8ug/ml for levofloxacin, 2 mg/L and 16 mg/L for antoflolxacin, 2 mg/L and 16 mg/L for ofloxacin. The Hunter-Gaston Discriminatory Index (HGDI) value for the 16-loci VNTR typing of M. intracellulare isolates was 0.994. VNTR differentiated the 150 isolates into 21 clusters and acquired a total of 121 unique patterns. Drug resistance profile was not independently associated with cluster strains. Conclusions: Clarithromycin, moxifloxacin and amikacin had the best antimicrobial activities in vitro against M. intracellulare isolates. The 16-loci VNTR typing revealed a highly discriminatory power and drug resistance profile was not independently associated with cluster strains.

Inhibition of sympathetic sprouting in CCD rats by lacosamide.

BACKGROUND: Early hyperexcitability activity of injured nerve/neuron is critical for developing sympathetic nerve sprouting within dorsal root ganglia (DRG) since lacosamide (LCM), an anticonvulsant, inhibits Na+ channel. The present study tried to test the potential effect of LCM on inhibiting sympathetic sprouting in vivo. METHODS: Lacosamide (50 mg/kg) was daily injected intraperitoneally into rats subjected to chronic compression DRG (CCD), an animal model of neuropathic pain that exhibits sympathetic nerve sprouting, for the 1st 7 days after injury. Mechanical sensitivity was tested from day 3 to day 18 after injury, and then DRGs were removed off. Immunohistochemical staining for tyrosine hydroxylase (TH) was examined to observe sympathetic sprouting, and patch-clamp recording was performed to test the excitability and Na+ current of DRG neurons. RESULTS: Early systemic LCM treatment significantly reduced TH immunoreactivity density in injured DRG, lowered the excitability level of injured DRG neurons and increased paw withdrawal threshold. These effects on reducing sympathetic sprouting, inhibiting excitability and suppressing pain behaviour were observed 10 days after the end of early LCM injection. In vitro 100 µmol/L LCM instantly reduced the excitability of CCD neurons via inhibiting Na+ current and reducing the amplitude of AP. CONCLUSIONS: All the findings suggest, for the first time, that early administration of LCM inhibited sympathetic sprouting and then alleviated neuropathic pain. SIGNIFICANCE: Early LCM administration inhibited sympathetic sprouting within DRG in CCD rats via reducing hyperexcitability of neurons. Early LCM administration suppressed neuropathic pain in CCD rats.

Trends in drug-resistant tuberculosis in China: data from a clinical tuberculosis centre.

OBJECTIVE: To describe the trends in prevalence of drug-resistant tuberculosis (TB) among in-patients in Beijing Chest Hospital, Beijing, China, using a 10-year retrospective study. DESIGN: From 2005 to 2014, 18 310 in-patients with TB were recruited for the study, most of whom were referrals; no distinction was made between new and previously treated cases. Drug susceptibility testing (DST) was performed in culture-positive cases using the proportion method to determine multidrug-resistant TB (MDR-TB) and extensively drug-resistant TB (XDR-TB). Risk factors associated with drug resistance were identified. RESULTS: A total of 5141 (28.0%) samples were culture-positive. DST results showed that 860 (16.7%) cases were MDR-TB and 176 (3.4%) were XDR-TB. MDR-TB and XDR-TB were detected in respectively 21.2% and 12.5% of new cases. The rate of MDR-TB and XDR-TB gradually increased from 2005, with MDR-TB reaching a peak in 2008 and XDR-TB in 2009. These data closely mirror national survey data on this region, patient age and occupation. CONCLUSION: Trends in MDR-TB and XDR-TB prevalence during the past decade and their inflection points were determined, which complemented reports from previous national surveys. This information is useful for fighting TB in China.

Water-soluble metal nanoparticles stabilized by plant polyphenols for improving the catalytic properties in oxidation of alcohols.

Plant polyphenols extracted from plants are one of the most abundant biomasses in nature, which are typical water soluble natural polymers. Herein, we reported a facile approach for the synthesis of platinum nanoparticle (PtNP) aqueous colloid by utilizing black wattle tannin (BWT, a typical plant polyphenol) as amphiphilic stabilizer. The phenolic hydroxyls of BWT provide the PtNPs with enough hydrophilicity, and their reduction ability could protect the PtNPs from deactivation caused by oxygen atmosphere. Additionally, the hydrophilic nature of BWT could efficiently promote the oxidation of alcohols in water, meanwhile, the hydrophobic and rigid backbones of plant polyphenols are able to suppress the PtNPs from aggregating, thus ensuring the high dispersion of the PtNPs during reactions. Under mild aerobic conditions, the as-prepared BWT-Pt colloid catalyst exhibited high activity in a series of biphasic oxidation of aromatic alcohols and aliphatic alcohols. As for the cycling stability, the BWT-Pt catalyst showed no obvious decrease during the 7 cycles, revealing superior cycling stability as compared with the counterparts using PVP or PEG as the stabilizer.

Effects of TNF-α on Cementoblast Differentiation, Mineralization, and Apoptosis.

Tumor necrosis factor-α (TNF-α) is involved in various inflammatory processes, including periodontitis. Although the influences of TNF-α on periodontal ligament fibroblasts and osteoblasts have been widely documented, its effects on cementoblasts, the cells responsible for cementum production, remain largely unknown. In this study, we found that TNF-α suppressed the mineralization ability of cementoblasts by inhibiting differentiation and inducing apoptosis. Various signaling pathways, such as p53, PP2AC, p38, Erk1/2, JNK, PI3K-Akt, and NF-κB, were activated during this process. The use of a specific inhibitor and siRNA transfection confirmed that the effects of TNF-α on differentiation and apoptosis in cementoblasts were partially abrogated by inhibiting p53 activity. By contrast, the effects of TNF-α were even exacerbated by the inhibition of the p38, Erk1/2, JNK, PI3K-Akt, and NF-κB pathways. Moreover, p53 activity was further enhanced by blocking the p38, Erk1/2, JNK, and PI3K-Akt signaling pathways. Taken together, these results suggested that the differentiation inhibition and apoptosis in cementoblasts induced by TNF-α were partially dependent on p53 activity. The p38, Erk1/2, JNK, PI3K-Akt, and NF-κB pathways were also activated but acted as balancing players to limit rather than conduct the negative effects of TNF-α. These balancing effects were dependent, or at least partially dependent, on p53, except for the NF-κB pathway.

Kidney transplantation from donors after brain or cardiac death in China--a clinical analysis of 94 cases.

BACKGROUND: We analyzed outcomes of kidney transplantation (KTx) from donation after brain death (DBD) or cardiac death (DCD) in China under the current level of the health care system. METHODS: Among 94 KTx performed from February 2007 to March 2012 in two organ transplant centers in the south of China, 50 KTx were used DBD and 44 DCD donors. We retrospectively analyzed the clinical outcomes. RESULTS: At a mean follow-up of 25.5 months, the 1-year and 2-year graft survival rates were 96.8% and 95.2% respectively. Delayed graft function (DGF) occurred in 27.7% recipients, three of whom lost graft function. Among six observed acute rejection episodes, five were reversed. When compared to the DCD group in DBD patients were apt to recover from DGF. Serum creatinine decreased more promptly in the DBD than in DCD group. Serum creatinine in the DCD group increased after months 12, when it was significantly higher than that in the DBD group (P < .05). CONCLUSIONS: Kidney transplantation from DBD donors showed good outcomes with few complications. Although KTx from DCD donors showed a higher DGF rate and longer duration of graft recovery, we achieved favorable short-term clinical outcome using this source.

Involvement of ventrolateral orbital cortex 5-HT 1-7 receptors in 5-HT induced depression of spared nerve injury allodynia.

The present study examined the roles of ventrolateral orbital cortex (VLO) 5-HT1, 5-HT2, 5-HT3, 5-HT4, 5-HT5, 5-HT6 and 5-HT7 receptor subtypes in mediating 5-HT-induced antiallodynic actions in the rat spared nerve injury (SNI) pain model. Changes in paw withdrawal threshold (PWT) were measured using von-Frey filaments. Microinjection of 5-HT (2, 5 and 10µg, in 0.5µl) into the VLO depressed allodynia induced by SNI, and the PWT increased in a dose-dependent manner. Microinjection of selective 5-HT1A, 5-HT2, 5-HT3, 5-HT4, 5-HT5A, 5-HT6 and 5-HT7 receptor antagonists, 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine hydrobromide (NAN-190) (10µg), cyproheptadine (50ng), granisetron hydrochloride (granisetron) (10µg), 1-[2-[(methylsulfonyl)-amino]ethyl]-4-piperidinyl]methyl1-methyl-1H-indole-3-carboxylate (GR113808) (5µg), SB699551 dihydrochloride (SB699551) (10µg), SB258585 dihydrochloride (SB258585) (2µg) or SB269970 hydrochloride (SB269970) (10µg) into the VLO 5-min prior to 5-HT (10µg) injection, all antagonized the 5-HT-induced inhibition of allodynia. In addition, these antagonists applied alone to VLO did not influence allodynia. These results suggest that although 5-HT1-7 receptor subtypes in the VLO do not have a tonic modulatory action on the allodynia induced by SNI, they are involved in mediating the depression of the SNI allodynia produced by injection of 5HT into VLO.

Metabotropic glutamate subtype 7 and 8 receptors oppositely modulate cardiac nociception in the rat nucleus tractus solitarius.

Recent study from our laboratory has indicated that microinjection of glutamate into the nucleus tractus solitarius (NTS) facilitates the cardiac-somatic reflex induced by pericardial capsaicin. Further, N-methyl-d-aspartate (NMDA) receptors and metabotropic glutamate receptors (mGluRs) mediate this function. However, the roles of the individual receptor subtypes or subunits in modulating cardiac nociception are unknown. Among the three groups of mGluRs, group III mGluRs are the primary mGluR subtype expressed in visceral afferent neurons in the NTS. The present study examined the roles of group III mGluRs and their subtype 7 and 8 receptors (mGluR7 and mGluR8) in modulating the cardiac-somatic reflex induced by pericardial capsaicin, which was monitored by recording electromyogram (EMG) activity from the spinotrapezius muscle in anesthetized rats. Intra-NTS microinjection of a group III mGluR agonist, l-(+)-2-Amino-4-phosphonobutyric acid (l-AP4, at 1, 10, and 20 nmol) or a selective mGluR7 agonist, N,N'-diphenylmethyl-1,2-ethanediamine dihydrochloride (AMN082, at 1, 2, and 4 nmol) both decreased the EMG response in a dose-dependent manner. This decrease was inhibited by the group III mGluR antagonist (RS)-α-Methylserine-O-phosphate (MSOP, at 20 nmol). In contrast, intra-NTS microinjection of a selective mGluR8 agonist, (S)-3, 4-dicarboxyphenylglycine (DCPG, at 6 and 8 nmol), significantly increased the EMG response above control levels. This effect was eliminated by intra-NTS MSOP and by vagal deafferentation. These data suggest that group III mGluRs and mGluR7 in the NTS display an inhibitory effect, while mGluR8 displays a facilitatory effect in modulating cardiac nociception, and this facilitatory effect is dependent on vagal afferents.

Chemical lesioning and glutamate administration reveal a major role for the nucleus tractus solitarius in the cardiac-somatic reflex in rats.

Many patients suffer from secondary muscle hyperalgesia after experiencing angina pectoris. In this study, we examined the role of the nucleus tractus solitarius (NTS) and glutamate receptors in modulating cardiac-evoked muscle hyperalgesia induced by pericardial capsaicin, which was monitored by recording electromyogram (EMG) activity from the spinotrapezius muscle in the anesthetized rat. Unilateral chemical lesioning of the commissural NTS with the neurotoxin ibotenic acid significantly depressed the cardiac-somatic reflex; the EMG responses decreased to 56.4 ± 6.9% of that of the controls (5 of 5). Microinjection of the excitatory amino acid glutamate, at 10, 20, and 50 nmol, into the commissural NTS increased the EMG response, in a dose-dependent manner, to 116.9 ± 4.9%, 143.9 ± 10.2%, and 214.2 ± 15.8% (n=8), respectively, of that of the controls. In contrast, microinjection of the N-methyl-D-aspartate (NMDA) receptor antagonist (+)-5-methyl-10, 11-dihydro-5H-dibenzo [a, d]-cyclohepten-5,10-imine maleate (MK-801) at 4 and 6 nmol, decreased the EMG response to 45.2 ± 10.6% and 36.8 ± 14.3%, respectively, of that of the controls (n=8 for each dose). Similarly, the metabotropic glutamate receptor (mGluR) antagonist (RS)-a-methyl-4-carboxyphenylglycine (MCPG), at 2.5 and 5 nmol, decreased the EMG response to 65.2 ± 16.3% and 57.0 ± 4.2%, respectively, of that of the controls. When a combination of MK-801 and MCPG was administrated, the EMG response further decreased to 22.5 ± 13.2% (n=6) of that of the controls. However, administration of a non-NMDA receptor antagonist 6, 7-dinitroquinoxaline-2, 3-dione (DNQX), at 2 and 5 nmol, had no effect on the EMG response. These results suggest that the NTS is involved in the facilitation of the cardiac-somatic reflex, and that the NMDA receptor and mGluRs play an important role in mediating this effect.

Cloning expression, monoclonal antibody preparation and serologic study of mammaglobin in breast cancer.

UNLABELLED: Mammaglobin may be a potential serum biomarker for the differential diagnosis of breast cancer. 260 serum samples were collected from 127 untreated breast cancer patients and 133 healthy volunteers to analyze the sera expression of mammaglobin and its implications for both. The expression vector of pGEX-4T-2-Mammaglobin and pBVIL1-Mammaglobin were constructed and transformed into E.coli.HB101 for expression. The mice were immunized with the purified recombinant protein to prepare monoclonal antibody and to detect by ELISA the serum of normal people and breast cancer patients. Recombinant mammaglobin antigen was effectively expressed in E.coli. Two hybridoma cell lines were obtained after the mice were immunized by pGEX-4T-2-mammaglobin. 133 cases of normal serum and 127 cases of breast cancer serum were analyzed by ELISA. The sera expression level of mammaglobin in breast cancer group (average OD value 0.645±0.223) was significantly (p KEYWORDS: mammaglobin; cloning expression; monoclonal antibody; serologic study; breast cancer.

The role of dopamine receptors in ventrolateral orbital cortex-evoked anti-nociception in a rat model of neuropathic pain.

The present study examined the role of dopamine and D(1)-and D(2)-like dopamine receptors in ventrolateral orbital cortex (VLO)-evoked anti-hypersensitivity in a rat model of neuropathic pain, as well as the possible underlying mechanisms. Results showed that microinjection of apomorphine [(R(-)-apomorphine hydrochloride)], a non-selective dopamine receptor agonist, into the VLO attenuated spared nerve injury (SNI)-induced mechanical allodynia in a dose-dependent manner. This effect was completely blocked by the D(2)-like dopamine receptor antagonist S(-)-raclopride(+)-tartrate salt (1.5 microg), but was enhanced by the D(1)-like dopamine receptor antagonist SCH23390 (R(+)-SCH-23390 hydrochloride, 5.0 microg). The attenuating effect of apomorphine on mechanical allodynia was mimicked by application of the D(2)-like dopamine receptor agonist quinpirole [((-)-quinpirole hydrochloride, 0.5, 1.0, and 2.0 microg)]. In addition, microinjection of larger doses (10 and 20 microg) of SCH23390 into the VLO significantly attenuated allodynia. Furthermore, microinjections of GABA(A) receptor antagonists, bicuculline [(+)-bicuculline,(S), 9(R)] and picrotoxin (200 and 300 ng for both drugs), into the VLO attenuated mechanical allodynia. A small dose of bicuculline or picrotoxin (100 ng) resulted in increased quinpirole (0.5 microg)-induced anti-allodynia. In contrast, GABA(A) receptor agonists, muscimol hydrochloride (250 ng) or THIP [(2,5,6,7-retrahydroisoxazolo(5,4-c)pyridine-3-ol hydrochloride, 1.0 microg)], blocked quinpirole (2.0 microg)-induced attenuation. These results suggest that the dopaminergic system is involved in mediating VLO-induced anti-hypersensitivity, activation of D(2)-like dopamine receptors, and inhibition of D(1)-like receptors resulting in anti-hypersensitivity. In addition, the mechanisms of GABAergic disinhibition might be involved in D(2)-like receptor mediating effects in neuropathic pain.

Simultaneous aortic and coronary assessment in abdominal aortic aneurysm patients by thoraco-abdominal 64-detector-row CT angiography: estimate of the impact on preoperative management: a pilot study.

OBJECTIVES: To estimate the influence of information on the coronary arteries obtained from routine thoraco-abdominal CT angiography (CTA) on pre-operative clinical management in abdominal aortic aneurysm (AAA) patients. METHODS: Twenty-eight AAA patients underwent pre-operative thoraco-abdominal electrocardiography (ECG)-gated 64-detector-row CTA to evaluate aortic pulsatility for prosthesis size matching. Retrospectively, the coronaries were reconstructed from the same data set and scored on a per segment basis for stenosis (0%, 50%) and grading confidence (poor, adequate or high). An experienced cardiologist was presented information on patient characteristics obtained from patient records and CTA findings. Suggested changes in European Society of Cardiology guidelines based patient management based on CTA information were scored. RESULTS: On CTA, 17 patients (61%) had significant coronary disease (>50% stenosis) including left main (n=4), single (n=7) and multiple (n=6) vessel disease. Grading confidence was adequate or high in 86% of proximal and middle segments. Based on CTA findings, patient management would have been changed in 4 out of the 28 patients (14%; 95% CI 1-27%) by adding coronary angiography (n=4). In five patients who underwent coronary artery bypass grafting previously, CT did not change management but confirmed graft patency. CONCLUSIONS: Information on coronary pathology and coronary bypass graft patency can be readily obtained from thoraco-abdominal CTA and may alter pre-operative patient management, as shown in 14% of AAA patients in our study.

Activation of mu-opioid receptors in thalamic nucleus submedius depresses bee venom-evoked spinal c-Fos expression and flinching behavior.

Previous studies have indicated that mu-opioid receptors in the thalamic nucleus submedius (Sm) are involved in descending antinociception in behavioral tests. The present study examined the effect of mu-opioid receptor activation in the Sm upon bee venom-evoked c-Fos expression in the spinal dorsal horn associated with flinching behavior, and determined whether the ATP-sensitive potassium channel (K-ATP channel) was involved in this effect in a rat model. A dilute bee venom solution, subcutaneously injected unilaterally into a rat hind paw pad, induced significant c-Fos expression in the lumbar spinal dorsal horn, which is associated with paw flinching behavior. This effect was depressed by microinjection of the mu-opioid receptor agonist [d-Ala2, N-MePhe4, Gly-ol5]-enkephalin (DAMGO) into the Sm, which was antagonized by pre-treatment with mu-receptor antagonist beta-funaltrexamine at the same Sm site. Further studies found that glibenclamide, a K-ATP channel inhibitor, also blocked DAMGO-induced inhibition. These results provide functional anatomic support for the involvement of Sm and mu-opioid receptors in the modulation of persistent inflammatory nociception, and suggest that these effects were produced by opening K-ATP channel and inhibiting neuronal activity. Together with previous studies, the inhibition of the neuronal activity induced by mu-opioid receptor activation may activate descending antinociceptive pathways through a GABAergic disinhibitory mechanism and depress the nociceptive information transmission at the level of the spinal cord.

The role of 5-HT receptor subtypes in the ventrolateral orbital cortex of 5-HT-induced antinociception in the rat.

The present study examined the involvement of 5-HT in the ventrolateral orbital cortex (VLO) on descending antinociception and determined which subtypes of 5-HT receptors mediated this effect. This study focused on the effects of 5-HT microinjection in the VLO of lightly anesthetized male rats on the radiant heat-evoked tail flick (TF) reflex, as well as the influence of 5-HT(1A), 5-HT(2), 5-HT(3), and 5-HT(4) receptor subtype antagonists on the effect of 5-HT. Results showed that 5-HT microinjection (2, 5, 10 microg, in 0.5 microl) into the VLO depressed the TF reflex in a dose-dependent manner. Pretreatment with 5-HT receptor antagonists (1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine hydrobromide (NAN-190), cyproheptadine hydrochloride (CPT) and 1-methyl-N-(8-methyl-8-azabicyclo[3.2.3]-oct-3-yl)-1H-indazole-3-carboxamide maleate salt (LY-278,584)), specific for 5-HT(1A), 5-HT(2) and 5-HT(3) receptors, respectively, partially reversed the 5-HT-evoked inhibition. In contrast, the 5-HT(4) receptor antagonist, 1-[2-[(methylsulfonyl)-amino]ethyl]-4-piperidinyl]methyl1-methyl-1H-indole-3-carboxylate (GR 113808), had no effect on the inhibition of 5-HT. Microinjections of NAN-190, CPT and LY-278,584 alone into the VLO had no effect on the TF reflex. These results suggest that 5-HT(1A), 5-HT(2) and 5-HT(3), but not 5-HT(4) receptors, are involved in mediating 5-HT-induced antinociception in the VLO. According to different properties and distribution patterns of the 5-HT receptor subtypes on neurons, the possible mechanism of 5-HT activation of the VLO-periaqueductal gray (PAG) descending antinociceptive pathway is discussed.