Rapid confirmation of five non-typhoidal Salmonella (NTS) serotypes using a low-cost portable-DIY OpenPCR© thermocycler

Aims@#OpenPCR is a low cost yet accurate thermocycler which can be self-built. The aim of the study is to highlight a low-cost alternative method for rapid confirmation of five predominant non-typhoidal Salmonella (NTS) serotypes using a multiplex PCR on a portable-DIY OpenPCR© thermocycler. @*Methodology and results@# Eight multiplex polymerase chain reaction (mPCR) samples containing genomic DNA of S. Agona selectively placed on the wells of the conventional PCR and OpenPCR© thermocyclers showed uniform heating in both thermocyclers. The limit of detection was similar for both thermocyclers for all five serotypes. The limit of detection for S. Typhimurium, S. Agona and S. Weltevreden was 10 pg/µL whereas the limit of detection for S. Enteriditis and S. Heidelberg was 1 pg/µL and 100 pg/µL, respectively. This assay incorporated a panel of unique genes; STM4495, SEN1392, SeHa-C4893, SeAg-B1096 and SENTW-3241 which were previously identified to be specific for S. Typhimurium, S. Enteritidis, S. Heidelberg, S. Agona, and S. Weltevreden, respectively, as well as the pan-Salmonella gene invA as internal control (IC) and pan-bacteria gene 16S rRNA to serve as amplification control (AC). The analytical specificity of the mPCR assay was found to be 100% for all five NTS using OpenPCR© thermocyclers. @*Conclusion, significance and impact of study@#The feasibility and low cost of the OpenPCR© thermocycler makes this device an ideal alternative for mPCR assay for rapid confirmation of NTS serotypes.

Expression of NTS in hepatocellular carcinoma(HCC)is associated with the formation of inflammatory microenvironment, more epithelial mesenchymal transition in cancer, and worse prognosis / 中国肿瘤临床

Objective:This work aims determine the expression of the neurotensin (NTS) gene in hepatocellular carcinoma (HCC) subgrouping using immunohistochemical staining (IHC) as well as to evaluate the correlation between the activation of NTS/IL-8 pathway in HCC and inflammatory response in microenvironment and epithelial mesenchymal transition (EMT) in cancer and in the prognosis of patients. Methods:Tumor tissues and corresponding adjacent normal tissue were collected from 64 cases of HCC patients. The expression levels of NTS protein and multiple inflammation and EMT-related proteins, including IL-8, VEGF, MMP9, CD68, E-Cadherin,β-Catenin, and Vimentin, were examined in 64 cases of paraffin-embedded HCC tissues using the immunohistochemistry (IHC) staining method. The clinical outcome and overall survival (OS) among 64 cases of HCC patients were compared. Results:We found that the frequency of NTS-expressing tissues among all HCC samples was 17.19%(11/64). Significantly increased IL-8 protein was confirmed in 90.91%of NTS+HCC samples and was positively correlated with the levels of NTS protein in cancer tissues (P=0.036), which implied the dysfunctional activation of NTS/IL-8 pathway in HCC. The levels of VEGF and MMP9 were significantly correlated with the co-expression of NTS and IL-8 in HCC. Evident features of EMT, including decreased membrane expression of E-Cadherin and increased accumulation of cytoplasmicβ-Catemin and Vimentin, were found in NTS+IL-8+samples. The co-expression of NTS and IL-8 in cancer was significantly correlated with the clinical outcomes of patients, as the mortality rate of NTS+IL-8+HCC patients is 2.5-fold higher than that of others after surgery (P=0.022).Accordingly, the OS of NTS+IL-8+HCC patients significantly decreased (24.65±4.45 m vs. 75.79±16.32 m, P=0.013), and these patients are at a higher risk of death at an expected hazard ratio (HR) of 3.457. Conclusion:The NTS/IL-8 pathway is dysfunctionally activated in a subgroup of HCC samples. Highly expressed NTS is associated with increased inflammatory response in microenvironment, enhanced EMT in cancer, and worse prognosis in HCC patients.

Treatment and Mechanism of Surface Electrical Stimulation for Severe Dysphagia Caused by Lower Brainstem Infarction / 中国康复理论与实践

@#Objective To study the potential mechanism of recovery from dysphagia after surface electrical stimulation. Methods 3 cases recovery from dysphagia caused by lower brainstem infarction after surface electrical stimulation of lower mandible and trigeminal nerves were analyzed. Results After 3～16 weeks surface electrical stimulation, the swallow assessment scores reached from 0 to 6 in all the 3 patients. Conclusion Surface electrical stimulation can facilitate the recovery of swallow function, which may be involved with the sensory input, especially the integration of nucleus of the solitary tract (NTS).

A Clinicomycologic Study of Dermatophytosis of the External Auditory Meatus / 대한피부과학회지

BACKGROUND: Otomycosis is a mycotic disease of the external auditory meatus. Aspergillus is the most common pathogen. Despite the fact that a great number of fungi are considered as the cause of otomycosis, otomycosis due to dermatophytes occurs with great rarity. OBJECTIVE: The purpose of this study was to investigate clinical and mycological features of dermatophytosis in the external auditory meatus. METHODS: We performed a clinical and mycologic analysis of 17 patients who had been clinically and mycologically diagnosed with dermatophytosis of the external auditory meatus. In addition, amplication of TRS-1 and TRS-2 of the ribosomal DNA nontranscribed spacer was performed on strains of Trichophyton (T.) rubrum and T. raubitshcekii which were isolated from skin lesions of the external ear and cerumen in 6 patients. RESULTS: The ratio of male to female patients was 1.4:1. The age of patients were 6 to 72 and mean age of onset is 36.4. Eleven patients had concomitant dermatophytosis of other sites. Tinea unguium was combined in 7 cases, tinea pedis in 4 cases, tinea corporis in 3 cases, tinea manus in 2 cases and tinea cruris in 1 case. The positive rate of KOH examination was 88.2% and the positive rate of fungal culture on PDAC media was 88.2%. The most common organism cultured was T. rubrum (80.0%), followed by T. raubitschekii (13.3%) and T. mentagrophytes (7.7%). Ten patients were treated with topical terbinafine and seven patients with oral itraconazole, so all patients were cleared of fungi from the external auditory meatus and inflammation was resolved. With TRS-1 and TRS-2 amplication, 4 of 5 pairs of T. rubrum and 3 strains of T. raubitshcekii showed the same types regardless of the site in each patient. CONCLUSION: Dermatophytosis occurs rarely in the external auditory meatus but we experienced 17 cases of dermatophytosis at this site. With suspected otomycosis, dermatophytosis of the external auditory meatus and concomitant dermatophytosis must be investigated and treated simultaneously to prevent any recurrence.

COMPARISON OF SUPPRESSIVE EFFECT BETWEEN THE LOW MOLECULAR WEIGHT NATURAL TUMOR SUPPRESSOR OF HUMAN FETAL HEPATOCYTE AND THAT OF EMBRYO CALF HEPATOCYTE ON THE GROWTH OF MURINE S- 180 CELLS / 肿瘤研究与临床

Objective:Effect of the low molecular weight natural tumor suppressor (LMW NTS) of human fetal hepatocyte and that of embryo calf hepatocyte on the growth of murine S 180 cells was observed.Methods:LMW NTS was isolated respectively from human fetal hepatocyte and embryo calf hepatocyte.(1)In vitro two layers agar plate culture method was used to detect the suppressive effect of the human fetal hepatocyte LMW NTS and the emtryo calf hepatocyte LMW NTS on the growth of murine S 180 cells.(2)Surviral of the mice bearing S 180 cells and treated respectively with intraperitoneal injections of the human fetal hepatocyte LMW NTS or the embryo calf hepatocyte LMW NTS(0.5 mg/one mouse/day for 21 days)was surveyed.Results:Both the human fetal hepatocyte LMW NTS and the embryo calf hepatocyte LMW NTS can obviously inhibit the growth of murine S 180 cell,and the suppressive effect was enhanced while the concentration of LMW NTS was increased.Survival of the mice bearing tumor that treated with intraperitoneal injections of the human fetal hepatocyte LMW NTS or the embryo calf hepatocyte LMW NTS was remarkably prolonged.Conclusion:The LMW NTS of embryo calf hepatocyte is similar to that of human fetal hepatocyte,and it possesses marked anti tumor activity both in vitro and in vivo.

Cardiovascular Effects of Endogenous GABA in the Nucleus Tractus Solitarius

BACKGROUND: The nucleus tractus solitarius (NTS), the region of the brain stem in which primary baroreceptor afferents teminate, is critically important in the normal regulation of arterial pressure (AP). In the NTS, excitatory amino acids such as L-glutamate serve as the main neurotransmitter in the regulation of AP. However, the function of GABA in the NTS has not been established. To test the function of GABA, we applied GABAergic agents to the NTS. METHODS: The experiments were conducted on adult male Sprague-Dawley rats weighing 300-500g. A cannula (PE-50 tubing filled with heparinized saline) was inserted into the femoral artery for recording of AP and heart rate(HR). Another cannula was inserted into the femoral vein for administration of nitroprusside or phenylephrine. After rats were placed on a sterotaxic instrument, the dorsal surface of the medulla was exposed, and with the aid of a surgical microscope, the NTS was visualized. Drug injections were made into the NTS using single- or three-barreled grass micropipettes pulled to an outer diameter of 80-100(micro)m and connected to a 1(micro)l Hamilton syringe. RESULTS: The follwing results were obtained in this experiment. Injection into the NTS of 10 or 20 nmol nipecitic acid, a selective inhibitor of GABA untake, produced an increase in AP. The pressor responses evoked by two doses of nipecotic acid were not significantly different. Injection of GABA(A) agonist, musciml(5 pmol in 80 nl artificial CSF) and GABA(B) agonist, baclofen (20 pmol in 80 nl) into the NTS of urethane-anesthetized rats prodused an increase in AP of 16.6+/-1.3 and 27.6+/-1.5 mmHg, respectively. Thus the pressor response to GABA(B) agonist was greater than to GABA(A) agonist. On the other hand, microinjection of GABA(A) antagonist, bicuculline and GABA(B) antagonist, phaclofen into the NTS decreased AP by approximately 13.4+/-1.0 and 20.9+/- mmHg, respectively. Thus injection of nipecotic acid into the NTS was greater in control group compared with the muscimiol or baclofen groups. The AP changes caused by i.v. injection of nitroprusside or phenylephrine were smallest in control group and greatest in the baclofen group. When calculated as baroreflex sensitivity, the change was greatest in control group and smallest in the baclofen group. CONCLUSION: From these results it was concluded that GABA in the NTS plays an important role in the regulation of AP, especially through GABA(B) receptors, and have an inhibitory effect on baroreceptor reflex.

Subdiaphragmatic vagotomy reduce the responses of fever and c-Fos expression in rat PVN and NTS to LPS / 中国免疫学杂志

Objective:To study the possibility that responses of fever and c Fos expression in rat PVN and NTS to intraperitoneal administration of LPS are mediated by vagal afferents.Methods:Rectal temperature was detected by digital temperature detecting instrument.c Fos expression was detected by immunohistochemistry staining.Results:The rectal temperature change value in vagotomy LPS group was significantly decreased compared with that in sham LPS group,and there was striking difference between them,P