Open peroral endoscopic myotomy for the treatment of achalasia: a case series of 82 cases.

During peroral endoscopic myotomy (POEM), creation of the tunnel is highly technically demanding and mucosal injury is one of the most common potential complications. We explored a method without a submucosal tunnel, which we call open peroral endoscopic myotomy (O-POEM). This study aimed to assess the feasibility and safety of O-POEM. O-POEM was performed on 82 patients with achalasia. Treatment success was defined as an Eckardt score of less than or equal to 3 after the myotomy. Adverse events including operative and postoperative adverse events were recorded. Treatment success and procedure-related adverse events were analyzed. After a median follow-up of 18 months (range: 6-26 months), the treatment success (Eckhart score ≤3) was achieved in 96.3% of cases (mean score pre- vs. post-treatment (7.4 vs. 1.8); P < 0.001) with a recurrence of 3 cases. Ten patients (12.2%) had adverse events consisting of 2 cases of mediastinitis, 1 case of post-O-POEM bleeding, 1 case of subcutaneous emphysema, 6 cases of pleural effusion. Two cases of mediastinitis required intraprocedural drainage, and other patients were managed by endoscopy and conservative medical treatment. There were no deaths. No patients required surgical conversion. Clinical reflux occurred in 15.9% of patients (13/82). O-POEM was reliable and effective for the treatment of achalasia. In addition, O-POEM might be a better option for patients with severe submucosal fibrosis.

Preparation and characterisation of andrographolide niosomes and its anti-hepatocellular carcinoma activity.

OBJECTIVE: In this study, a novel andrographolide (AG) preparation formulation, niosomes, was prepared to improve the bioavailability and tissue distribution of AG. METHODS: The niosomal formulation of AG was prepared by film hydration/sonication method and tissue distribution was measured by liquid chromatography-mass spectrometry (LC-MS) method in mice, and anti-hepatocellular carcinoma (anti-HCC) activity was examined by MTT method in HepG2. RESULTS: Entrapment efficiency, drug-loading ratio and average particle size of AG niosomes were 72.36%, 5.90% and 206 nm, respectively. The tissue distribution in mice demonstrated that the AG niosomes were absorbed in liver much more than the free AG. Furthermore, the anti-HCC activity in HepG2 cells showed that there was no significant difference between free AG and AG niosomes. CONCLUSION: The present results suggest that AG niosomes may have a significant potential of liver targeting, which is valuable in chemotherapy of HCC.

Honey combined with silver ion dressing in complicated chronic wound of the scalp: a case report.

OBJECTIVE: Scalp wound healing is a complex process. Nonhealing wounds can become chronic wounds, which increase the trauma and economic burden on a patient and may even cause death in severe cases. Thus, it can be difficult to find an effective treatment for chronic wounds of the scalp. CASE PRESENTATION: We present a case of a 13-year-old female patient with a chronic wound caused by a scalp incision infection 3 months after two operations for craniotomy for arachnoid cyst resection and cystic lesion-cisterna magna drainage. After repeated dressing changes and two debridement operations, the incision had still not healed during the following year. The wound finally healed after 6 months of dressing changes by combining honey with silver ion dressings, and the incision had not re-ruptured after 10 months of follow-up. CONCLUSIONS: Honey combined with silver may be an effective method for the treatment of chronic scalp wounds.

[Effectiveness of the "14 plus 7 day quarantine" and "nucleic acid plus total antibody testing" strategy for screening imported patients with COVID-19 in Xiamen].

Objective: To analysis effectiveness of the "14 plus 7 day quarantine" and "nucleic acid plus total antibody testing" strategy (combined screening strategy) for screenin the imported patients with COVID-19 in Xiamen. Methods: The study populations were overseas travelers arriving in Xiamen from March 17 to December 31, 2020, and overseas travelers who had quarantine outside Xiamen for less than 21 days from July 18 to December 31, 2020. Data were collected and analyzed on the timing of detection, pathways, and test results of the imported patients with COVID-19 after implementing combined screening strategy. Results: A total of 304 imported patients with COVID-19 were found from 174 628 overseas travelers and 943 overseas travelers from other cities. A total of 163 cases (53.6%) were diagnosed by multitime, multisite intensive nucleic acid testing after positive finding in total antibody testing. Among them, 27 (8.9%) were first positive for nucleic acid in 14 plus 7 day quarantine and 136 were first positive for nucleic acid in 14-day quarantine. Only 8 of these individuals were tested positive for nucleic acid after positive total antibody testing. The other 128 individuals were tested positive for nucleic acid after being negative for average 2.3 times (maximum of 6 times). Aditional 155 cases might be detected by using the combined "14 plus 7 day quarantine" and " nucleic acid plus total antibody testing" strategy compared with "14-day quarantine and nucleic acid testing" strategy, accounting for 51.0% of the total inbound infections. So the combined screening strategy doubled the detection rate for imported patients with COVID-19. No second-generation case caused by overseas travelers had been reported in Xiamen as of February 26, 2021. Conclusions: Xiamen's combined screening strategy can effectively screen the imported patients with COVID-19 who were first positive for nucleic acid after 14 day quarantine. Compared with "14 day quarantine and nucleic acid testing", the combined screening strategy improved detection rate and further reduced the risk of the secondary transmission caused by the imported patients with COVID-19.

Spatially controlled synthesis of silver nanoparticles and nanowires by photosensitized reduction.

The present paper reports on the spatially controlled synthesis of silver nanoparticles (NPs) and silver nanowires by photosensitized reduction. In a first approach, direct photogeneration of silver NPs at the end of an optical fiber was carried out. Control of both size and density of silver NPs was possible by changing the photonic conditions. In a further development, a photochemically assisted procedure allowing silver to be deposited at the surface of a polymer microtip was implemented. Finally, polymer tips terminated by silver nanowires were fabricated by simultaneous photopolymerization and silver photoreduction. The silver NPs were characterized by UV-visible spectroscopy and scanning electron microscopy.

Quantitative determination of atorvastatin and ortho-hydroxy atorvastatin in human plasma by liquid chromatography tandem mass spectrometry and pharmacokinetic evaluation.

A specific, sensitive and simple method was developed to determine the levels of both atorvastatin and ortho-hydroxy atorvastatin in human plasma. The analytes and internal standard pitavastatin were extracted from plasma by liquid-liquid extraction, separated on a Zorbax SB-C18 column, eluted with a mobile phase of water:acetonitrile (45:55 v/v), both containing 5% methanol and 0.01% formic acid. Detection was performed with an electrospray ionization triple quadrupole mass spectrometer in positive ion mode using multiple reaction monitoring. The standard calibration curves of atorvastatin and ortho-hydroxy atorvastatin were linear in the concentration range of 0.2-20 and 0.1-20 ng/mL, respectively. The intra- and inter-day precisions were < 7.7% and the accuracy was within ± 5.9%. The method has been successfully used for the study of the pharmacokinetics of atorvastatin and ortho-hydroxy atorvastatin in Chinese patients with coronary heart disease after a single oral dose of 20 mg atorvastatin. The mean values for the area under the plasma concentration-time curve for atorvastatin and ortho-hydroxy atorvastatin were 63.1 and 46.9 ng.h/mL, respectively.

No infectious risk of COVID-19 patients with long-term fecal 2019-nCoV nucleic acid positive.

OBJECTIVE: It has recently been reported that some COVID-19 patients have long-term positive fecal nucleic acid after discharging from the hospital with negative nucleic acid in the respiratory tract, but it is unclear whether COVID-19 patients with positive long-term fecal nucleic acid tests have the risk of self-infection. PATIENTS AND METHODS: From January 25, 2020 to March 9, 2020, 5 COVID-19 patients with negative respiratory tract nucleic acid and positive fecal nucleic acid were observed and studied to explore whether these patients can re-infect themselves. Five patients with COVID-19 accompanied by diarrhea as the main gastrointestinal symptoms were carefully observed through clinical symptoms, imaging and other auxiliary examinations. The RT-PCR technology was used to continuously detect fecal and respiratory viral nucleic acids. The IgM antibody was detected on the 7th day of admission and IgM/IgG at the time of discharge. RESULTS: All 5 patients had symptoms of fever and diarrhea upon admission. The fecal nucleic acid was positive, as well as the throat swab was positive. All COVID-19 patients had positive IgM antibodies on the 7th day of admission and positive IgM and IgG at the time of discharge, and there were no abnormalities in the gastrointestinal examination on discharge. All 5 fecal nucleic acid tests were positive at the time of discharge. After continuous dynamic follow-up for 3-15 days, no clinical symptoms recurred, and the last nucleic acid test was negative. CONCLUSIONS: There is no risk of self-infection for COVID-19 patients with long-term 2019-nCoV nucleic acid positive in feces.

Negative viral nucleic acid test of induced sputum: an additional criteria for COVID-19 patient's discharge.

OBJECTIVE: Concerns have been raised that patients with Coronavirus Disease 2019 (COVID-19) are still infectious with a re-positive nucleic acid test of the pharyngeal swab after hospital discharge. The aim of this study was to investigate the clinical relevance of induced sputum as an additional indicator for the current clinical discharge criteria of COVID-19 patients to prevent virus recurrence. PATIENTS AND METHODS: Twenty-one COVID-19 patients who met the national clinical discharge criteria were discharged from the hospital and tested daily for the presence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) nucleic acid in their pharyngeal swabs and every other day for the presence of SARS-CoV-2 in their induced sputum. Once the patient's induced sputum was negative after two consecutive tests, testing was discontinued. RESULTS: Among 21 discharged patients from COVID-19, the first pharyngeal swab and induced sputum tests for viral nucleic acid were positive in 3 (14.3%) and 8 (38.1%) patients respectively. Induced sputum was significantly more positive than pharyngeal swab (p < 0.05). In our cohort, all pharyngeal swabs became negative at day 7, and all induced sputa turned negative at day 11 after discharge. Interestingly, patients with negative pharyngeal swabs experienced viral relapse, whereas patients with negative induced sputum did not revert to positivity. CONCLUSIONS: The detection rate of positive viral nucleic acid in induced sputum was high. Patients with negative induced sputum nucleic acid tests did not have a relapse of SARS-COV-2, indicating that viral nucleic acid testing of induced sputum should be used as an additional criterion for patients with national clinical discharge criteria COVID-19.

Circ-CCDC66 accelerates proliferation and invasion of gastric cancer via binding to miRNA-1238-3p.

OBJECTIVE: The aim of this study was to examine the expression of circ-CCDC66 in gastric cancer (GC) tissues and cell lines, as well as its correlation with the prognosis of GC. Moreover, the regulatory effects of circ-CCDC66 on biological behaviors of GC cells and its molecular mechanism were explored. PATIENTS AND METHODS: The relative expression level of circ-CCDC66 in GC tissues and cell lines was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between the circ-CCDC66 level and overall survival of GC patients was analyzed as well. The potential influences of circ-CCDC66 on proliferative and invasive abilities of GC cells were evaluated through 5-Ethynyl-2'-deoxyuridine (EdU), colony formation and transwell assay, respectively. Meanwhile, the cell cycle progression and apoptosis of GC cells affected by circ-CCDC66 were determined. In addition, the direct target miRNA of circ-CCDC66 was predicted and verified by bioinformatics method and Dual-Luciferase reporter gene assay, respectively. RESULTS: Circ-CCDC66 was significantly up-regulated in GC tissues and cell lines. Up-regulation of circ-CCDC66 indicated markedly worse prognosis of GC patients. Transfection of circ-CCDC66-siRNA remarkably attenuated proliferative and invasive abilities of BGC-823 and MGC-803 cells. Besides, GC cells were arrested in the G0/G1 phase, and the apoptotic rate was remarkably elevated after circ-CCDC66 knockdown. The Dual-Luciferase reporter gene assay verified that circ-CCDC66 bind to miRNA-1238-3p by competing with LHX2 (LIM-homeobox domain 2). MiRNA-1238-3p was significantly down-regulated in GC cells, whereas LHX2 was up-regulated. Furthermore, overexpression of miRNA-1238-3p in GC cells markedly suppressed the LHX2 level. CONCLUSIONS: Circ-CCDC66 is highly expressed in GC tissues and cell lines. Knockdown of circ-CCDC66 attenuates proliferative and invasive abilities of GC cells. Our results indicate that circ-CCDC66/miRNA-1238-3p/LHX2 axis may be a promising target for GC treatment.

Long non-coding RNA LINC00628 suppresses the growth and metastasis and promotes cell apoptosis in breast cancer.

OBJECTIVE: Breast cancer is the most common malignant tumor in women. However, the detailed mechanisms of its tumorigenesis remain largely unknown. Evidence and data have shown that abnormality in expression of Long non-coding RNA (LncRNA) is closely related to tumorigenesis. The aim of this study is to identify the detailed mechanisms of LncRNA LINC00628 in breast cancer. PATIENTS AND METHODS: The expression of LINC00628 in breast cancer tissues, adjacent non-cancerous tissues and cell lines were detected by qRT-PCR. Kaplan-Meier method and log-rank test were applied to analyze the overall survival of patients with low and high expression level of LINC00628 respectively. The LCC2 and MCF-7 cells were transfected with LINC00628 and the proliferation, invasion and migration were examined. The cell cycle distribution and cell apoptosis rate in LCC2 and MCF-7 cells after transfection with LINC00628 were explored by flow cytometry. The relative expression level of Bcl-2, Bax and Caspase-3 protein in LCC2 and MCF-7 cells after transfection with LINC00628 was detected by Western blotting. RESULTS: The relative expression level of LINC00628 in breast cancer tissues and cell lines were significantly decreased and the low expression level of LINC00628 has a poorer prognosis and lower overall survival rate. The overexpression of LINC00628 suppressed breast cancer cells proliferation, invasion and migration. Further, with the overexpression of LINC00628, cell cycle was arrested in G0/G1 phase in breast cancer cells and cell apoptosis was promoted. The relative expression of Caspase-3 and Bax protein were significantly increased and the relative expression of Bcl-2 protein was significantly decreased after transfection with LINC00628. CONCLUSIONS: The expression of LINC00628 was decreased in breast cancer. The overexpression of LINC00628 suppressed the proliferation, invasion and migration of breast cancer cells and promoted cell apoptosis associated with the regulation of Bcl-2/Bax/Caspase-3 signal pathway.

Rectification and rapid activation at low Ca2+ of Ca2+-activated, voltage-dependent BK currents: consequences of rapid inactivation by a novel beta subunit.

A family of accessory beta subunits significantly contributes to the functional diversity of large-conductance, Ca(2+)- and voltage-dependent potassium (BK) channels in native cells. Here we describe the functional properties of one variant of the beta subunit family, which confers properties on BK channels totally unlike any that have as yet been observed. Coexpression of this subunit (termed beta3) with Slo alpha subunits results in rectifying outward currents and, at more positive potentials, rapidly inactivating ( approximately 1 msec) currents. The underlying rapid inactivation process results in an increase in the apparent activation rate of macroscopic currents, which is coupled with a shift in the activation range of the currents at low Ca(2+). As a consequence, the currents exhibit more rapid activation at low Ca(2+) relative to any other BK channel subunit combinations that have been examined. In part because of the rapid inactivation process, single channel openings are exceedingly brief. Although variance analysis suggests a conductance in excess of 160 pS, fully resolved single channel openings are not observed. The inactivation process results from a cytosolic N-terminal domain of the beta3 subunit, whereas an extended C-terminal domain does not participate in the inactivation process. Thus, the beta3 subunit appears to use a rapid inactivation mechanism to produce a current with a relatively rapid apparent activation time course at low Ca(2+). The beta3 subunit is a compelling example of how the beta subunit family can finely tune the gating properties of Ca(2+)- and voltage-dependent BK channels.

Inactivation of BK channels mediated by the NH(2) terminus of the beta3b auxiliary subunit involves a two-step mechanism: possible separation of binding and blockade.

A family of auxiliary beta subunits coassemble with Slo alpha subunit to form Ca(2)+-regulated, voltage-activated BK-type K(+) channels. The beta subunits play an important role in regulating the functional properties of the resulting channel protein, including apparent Ca(2)+ dependence and inactivation. The beta3b auxiliary subunit, when coexpressed with the Slo alpha subunit, results in a particularly rapid ( approximately 1 ms), but incomplete inactivation, mediated by the cytosolic NH(2) terminus of the beta3b subunit (Xia et al. 2000). Here, we evaluate whether a simple block of the open channel by the NH(2)-terminal domain accounts for the inactivation mechanism. Analysis of the onset of block, recovery from block, time-dependent changes in the shape of instantaneous current-voltage curves, and properties of deactivation tails suggest that a simple, one step blocking reaction is insufficient to explain the observed currents. Rather, blockade can be largely accounted for by a two-step blocking mechanism (C(n) <---> O(n) <---> O(*)(n) <---> I(n)) in which preblocked open states (O*(n)) precede blocked states (I(n)). The transitions between O* and I are exceedingly rapid accounting for an almost instantaneous block or unblock of open channels observed with changes in potential. However, the macroscopic current relaxations are determined primarily by slower transitions between O and O*. We propose that the O to O* transition corresponds to binding of the NH(2)-terminal inactivation domain to a receptor site. Blockade of current subsequently reflects either additional movement of the NH(2)-terminal domain into a position that hinders ion permeation or a gating transition to a closed state induced by binding of the NH(2) terminus.

Gating properties conferred on BK channels by the beta3b auxiliary subunit in the absence of its NH(2)- and COOH termini.

Both beta1 and beta2 auxiliary subunits of the BK-type K(+) channel family profoundly regulate the apparent Ca(2)+ sensitivity of BK-type Ca(2)+-activated K(+) channels. Each produces a pronounced leftward shift in the voltage of half-activation (V(0.5)) at a given Ca(2)+ concentration, particularly at Ca(2)+ above 1 microM. In contrast, the rapidly inactivating beta3b auxiliary produces a leftward shift in activation at Ca(2)+ below 1 microM. In the companion work (Lingle, C.J., X.-H. Zeng, J.-P. Ding, and X.-M. Xia. 2001. J. Gen. Physiol. 117:583-605, this issue), we have shown that some of the apparent beta3b-mediated shift in activation at low Ca(2)+ arises from rapid unblocking of inactivated channels, unlike the actions of the beta1 and beta2 subunits. Here, we compare effects of the beta3b subunit that arise from inactivation, per se, versus those that may arise from other functional effects of the subunit. In particular, we examine gating properties of the beta3b subunit and compare it to beta3b constructs lacking either the NH(2)- or COOH terminus or both. The results demonstrate that, although the NH(2) terminus appears to be the primary determinant of the beta3b-mediated shift in V(0.5) at low Ca(2)+, removal of the NH(2) terminus reveals two other interesting aspects of the action of the beta3b subunit. First, the conductance-voltage curves for activation of channels containing the beta3b subunit are best described by a double Boltzmann shape, which is proposed to arise from two independent voltage-dependent activation steps. Second, the presence of the beta3b subunit results in channels that exhibit an anomalous instantaneous outward current rectification that is correlated with a voltage dependence in the time-averaged single-channel current. The two effects appear to be unrelated, but indicative of the variety of ways that interactions between beta and alpha subunits can affect BK channel function. The COOH terminus of the beta3b subunit produces no discernible functional effects.

[Ion channels in rat pancreatic beta cells].

Using perforated and cell-attached patch clamp techniques, the characteristics of ATP sensitive K(+) channels (K(ATP)), delayed rectifier K(+) channels (K(DR)), Ca(2+) and Na+ channels on single rat pancreatic beta cell membranes were studied. The results showed that (1) the efflux and influx conductance of K(ATP) channels was about 31 and 65 pS respectively, and the reversal potential of K(ATP) was about 60 mV; (2) K(DR) was activated completely after a latency of 20 ms, and K(DR) was about 1/3 of K(ATP); (3) whole cell Ca(2+) current reached a peak (40 60 pA) at 0 mV; L-type Ca(2+) channel was the main Ca(2+) channel in beta cells, but other types of high voltage activated Ca(2+) channels existed as well; and 4) whole cell Na(+) current reached a peak (200 400 pA) at 10 mV; but the expression level of Na(+) channel in beta cells varied among the cells. About half of the beta cells virtually had no Na(+) currents.

[Role of M-type receptor in internal calcium release and quantal secretion in rat adrenal chromaffin cells].

In rat single adrenal chromaffin cells, the effects of methacholine (MCh) on [Ca2+]i and catecholamine secretion were studied with fura-2 fluorescence and carbon fiber electrodes. In the presence of 2 mmol/L Ca2+ in the bath, locally applying 1 mmol/L MCh, either containing or not containing Ca2+, evoked both [Ca2+]i and secretion signals. In the absence of Ca2+ in the bath, MCh could still evoke [Ca2+]i and secretion. These results suggest that MCh causes release of Ca2+ from Ca2+ stores, which is sufficient to yield the evoked secretion. The Ca2+ store can be depleted by single MCh puff in the absence of Ca2+ in the bath.

Abnormal information processing in dementia of Alzheimer type. A study using the event-related potential's field.

Electrical field changes of event-related potentials (ERPs) were investigated in 26 patients with dementia of Alzheimer's type (DAT) and 12 age-matched normal subjects. The patients were assessed with the Clinical Dementia Rating and Mini-Mental State. Each patient selected had only mild to moderate mental disability. Auditory oddball stimulation was presented at 1.5 s intervals and 1,000 Hz for the nontarget and 2,000 Hz for the target tones, both at 85 dB. The target tones were 20% of all the tones. The reference-independent data (latency, global field power: GFP, dissimilarity index: DISS and location of centroids) were obtained and analyzed for each ERP component. The momentary electric strength or 'hilliness' of the ERPs landscape was indicated by GFP. The patients showed prolonged latencies and decreased P300 GFP amplitudes and of N100 GFP. These findings suggest that the abnormal electrical field of ERP may reflect abnormal information processing following the attentional process for target stimuli in DAT patients.