Laparoscopic transcystic common bile duct exploration: surgical indications and procedure strategies.

BACKGROUND: LTCBDE combined with or without modified techniques is safe and efficacious for the management of gallstones and concomitant, even large, common bile duct (CBD) stones. METHODS: To describe the surgical indications and procedure strategies of laparoscopic transcystic common bile duct exploration (LTCBDE), a retrospective analysis of 205 patients with concomitant gallstones and CBD stones treated using LTCBDE between June 2008 and June 2015 was performed. Clinical data on disease characteristics, methods for cystic duct incision and CBD stone extraction (with or without laser lithotripsy), and surgical outcomes were collected and reviewed. RESULTS: CBD stones were successfully cleared in all patients. No patient was converted to choledochotomy or laparotomy. The cystic duct diameter ranged 3-8 mm, and 85 patients with cystic duct diameter ≥ 5 mm. The mean time for CBD stone extraction was 25.3 min, with the operative time ranged from 63 to 170 min. Lithotripsy was used in 74 (36.1%) patients among which 26 patients with cystic duct diameter ≥ 5 mm. Estimated blood loss during surgery was 10-120 ml per patient, and no intra-operative blood transfusions were needed. The mean postoperative hospital stay was 5.1 (range 3-7) days, and postoperative complications developed in seven patients. No bile duct injury, stricture, remnant, recurrent stones, or other adverse events were observed during the mean follow-up of 8 months. CONCLUSIONS: Based on preoperative MRCP and intra-operative IOC findings about cystic duct diameter, the diameter of CBD, CBD stone size, we summarized and proposed the surgical indications and suitable techniques and strategies during LTCBDE.

Flight performance of Macdunnoughia crassisigna (Lepidoptera: Noctuidae).

Macdunnoughia crassisigna Warren (Lepidoptera: Noctuidae) is a highly destructive herbivore that poses a serious risk to cotton, maize, soybean, and cruciferous vegetables in East Asia. Examining the effects of various biotic and abiotic factors on the flight performance of M. crassisigna is crucial for a better understanding of its trans-regional migration. In this study, the flight activity of M. crassisignai moths of different ages, under different temperatures and relative humidity (RH) levels, was evaluated by tethering individuals to computerized flight mills for a 24-h trial period. The results showed that M. crassisignai had the capacity for sustained flight and the flight ability was strongest in 3-day-old individuals, and then their flight performance decreased significantly in older moths. For both sexes, temperature had a significant effect on their flight performance, and the flight activity was relatively higher at 24-28°C than other temperatures. There was a significant effect of RH on all flight parameters of the tested moths, and the flight activity was relatively higher at RH of 60-75% than other RH levels. For 3-day-old moths under the optimum conditions (24°C and 75% RH) throughout the 24 h scotophase, their mean flight distance reached 66 km, and the mean flight duration reached 13.5 h, suggesting M. crassisigna possess strong potential to undertake long-distance migration. These findings will be helpful for developing sound forecasting systems of this pest species.

Is Heliothis viriplaca (Lepidoptera: Noctuidae) a long-distance migrant?

Heliothis viriplaca (Hüfnagel) (Lepidoptera: Noctuidae) is an important polyphagous pest of beans, cotton, maize, and alfalfa worldwide. H. viriplaca is a long-distance migrant, and if so, what pattern of seasonal migration this species exhibits in northern China remains unknown. In this study, in order to determine the seasonal migration of H. viriplaca in northern China, the combination of searchlight trapping and ovarian dissection was carried out on an isolated small island in the center of the Bohai Strait during 2003-2014. The results confirmed that H. viriplaca undertakes long-distance migration on the prevailing winds of the East Asian monsoon airflows. This species exhibited a regular pattern of seasonal migration across the sea from May to October, but there was considerable yearly and monthly variation in the trapped numbers, with the majority being trapped in summer (67.99 ± 6.54%). The mean period when migration was detectable at the island was 116.5 ± 5.6 days from 2003 to 2014, with the shortest time span of 74 days in 2013 and the longest of 144 days in 2005. Trapped females in May and June showed a relatively higher mating rates and some degree of ovarian development when compared with July, August and September, suggesting the migration of this species is not completely bound by the 'oogenesis-flight syndrome'. These findings will be helpful to improve the forecasting system and managing strategies of H. viriplaca.

Seasonal migration of Cnaphalocrocis medinalis (Lepidoptera: Crambidae) over the Bohai Sea in northern China.

The rice leaf roller, Cnaphalocrocis medinalis (Guenée), is a serious insect pest of rice with a strong migratory ability. Previous studies on the migration of C. medinalis were mostly carried out in tropical or subtropical regions, however, and what the pattern of seasonal movements this species exhibits in temperate regions (i.e. Northern China, where they cannot overwinter) remains unknown. Here we present data from an 11-year study of this species made by searchlight trapping on Beihuang Island (BH, 38°24'N; 120°55'E) in the centre of the Bohai Strait, which provides direct evidence that C. medinalis regularly migrates across this sea into northeastern agricultural region of China, and to take advantage of the abundant food resources there during the summer season. There was considerable seasonal variation in number of C. medinalis trapped on BH, and the migration period during 2003-2013 ranged from 72 to 122 days. Some females trapped in June and July showed a relatively higher proportion of mated and a degree of ovarian development suggesting that the migration of this species is not completely bound by the 'oogenesis-flight syndrome'. These findings revealed a new route for C. medinalis movements to and from Northeastern China, which will help us develop more effective management strategies against this pest.

Expression of perilipin 2 (PLIN2) in porcine oocytes during maturation.

Perilipins have been reported to limit the interaction of lipases with neutral lipids within the droplets, thereby regulating neutral lipid accumulation and utilization. This study aimed to identify the location and expression of PLIN1 and PLIN2 in porcine oocytes during maturation. Quantitative real-time polymerase chain reaction (qRT-PCR), immunostaining and Western blot methods were used to characterize the expression and distribution patterns of PLIN1 and PLIN2 in porcine oocytes. The results showed that PLIN1 was not detectable in porcine oocytes. PLIN2 and BODIPY 493/503-detected neutral lipid droplets appeared identical distribution patterns and extensive colocalization in both GV and MII porcine oocytes. PLIN2 protein expression was higher in GV oocytes than that in MII oocytes (p < 0.05), although PLIN2 mRNA expression was similar in both groups. These findings suggested that PLIN2 was a major lipid droplet-associated protein in porcine oocytes.

Recruitment of GABA(A) receptors in chemoreceptor pulmonary neuroepithelial bodies by prenatal nicotine exposure in monkey lung.

Pulmonary neuroepithelial bodies (NEB) act as airway oxygen sensors and produce serotonin, a variety of neuropeptides and are involved in autonomic nervous system control of breathing, especially during the neonatal period. We now report that NEB cells also express a GABAergic signaling loop that is increased by prenatal nicotine exposure. In this study, cultured monkey NEB cells show hypoxia-evoked action potentials and hypoxia-sensitive K(+) current. As shown by both immunofluorescence and RT-PCR, monkey NEB cells synthesize and contain serotonin. The monkey NEB cells express the beta2 and beta3 GABA_A receptor subunits, GAD and also express alpha7, alpha4 and beta4 nicotinic receptor (nAChR) subunits. The alpha7 nAChR is co-expressed with GAD in NEB. The numbers of NEB and beta3 GABA_A receptor subunits expressed in NEB cells in lungs from control newborn monkeys were compared to lungs from animals that received nicotine during gestation. Prenatal nicotine exposure increased the numbers of NEB by 46% in lung and the numbers of NEB cells expressing GAD and GABA_A beta3 receptors increased by 67% and 66%, respectively. This study suggests that prenatal nicotine exposure can modulate NEB function by increasing the numbers of NEB cells and by increasing both GAD expression and beta3 GABA_A receptor subunit expression. The interaction of the intrinsic GABAergic system in the lung with nicotinic receptors in PNEC/NEB may provide a mechanism to explain the link between smoking during pregnancy and SIDS.

Reversal of diabetes in BB rats by transplantation of encapsulated pancreatic islets.

Prolonged survival of pancreatic islet allografts implanted in diabetic BB rats was achieved by encapsulation of individual islets in a protective biocompatible alginate-polylysine-alginate membrane without immunosuppression. Intraperitoneal transplantation of the encapsulated islets reversed the diabetic state of the recipients within 3 days and maintained normoglycemia for 190 days. Normal body weight and urine volume were maintained during this period, and no cataracts were detected in the transplant recipients. In contrast, control rats receiving transplants of unencapsulated islets experienced normoglycemia for less than 2 wk. These results demonstrated that microencapsulation can protect allografted islets from both graft rejection and autoimmune destruction without immunosuppression in an animal model that mimics human insulin-dependent diabetes.

Synthesis and Electrochemical Properties of Nitrogen-Doped Graphene (NG)/Nickle Oxide (NiO) Nanocomposite for Supercapacitor.

A novel nitrogen-doped graphene (NG)/nickle oxide (NiO) nanocomposite was synthesized by a facile two-step method, where NiO particles were dispersed on the surface of NG. The NG/NiO nanocomposite is characterized by using field-emission scanning electron microscopy (FE-SEM), transmission electron microscope (TEM), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD) and Raman spectroscopy. The electrochemical properties of NG/NiO nanocomposite have been studied using cyclic voltammetry (CV), galvanostatic charge/discharge and electrochemical impedance spectroscopy (EIS) techniques. Compared with the nitrogen-doped graphene, the electrode prepared by NG/NiO nanocomposite has a mass specific capacitance of 342 F g(-1) at scan rate of 5 mV s(-1), which is much higher than that of nitrogen-doped graphene (NG). The galvanostatic charge/discharge results show this new kind nanocomposite has high specific capacitance with 320 F g(-1) in the range of 0.1-0.5 V at a current density of 1 A g(-1). The enhanced supercapacitive performance of NG/NiO nanocomposite suggesting its promising potential in supercapacitors.

Microencapsulated parathyroid cells as a bioartificial parathyroid. In vivo studies.

Parathyroid cells were isolated from healthy rats, encapsulated in alginate-polylysine membranes, and injected intraperitoneally into rats on which total parathyroidectomies had been performed. Three days posttransplant, serum calcium and PTH-M concentrations had increased to near-normal levels in the recipient animals. Similar results were observed in a separate group of parathyroidectomized rats 3 days after free parathyroid cells were implanted, but within 4 weeks serum calcium and PTH-M concentrations had decreased almost to pretransplant levels in these rats. In the rats with encapsulated cell transplants, by contrast, serum calcium and PTH-M levels were significantly higher, even after 8 weeks. No therapeutic effects were observed in rats injected with empty capsules or in the control group, which received no capsules or cells. These results indicate that transplants of microencapsulated parathyroid cells can temporarily reverse aparathyroidism in rats without the use of immunosuppressive drugs, and that further studies are warranted to investigate possible future clinical applications of this treatment.

Synaptic excitation in the dorsal nucleus of the lateral lemniscus: whole-cell patch-clamp recordings from rat brain slice.

The synaptic events underlying the excitation of neurons in the rat's dorsal nucleus of the lateral lemniscus were studied by whole-cell patch-clamp recordings in a brain slice preparation of the auditory midbrain. Both current-clamp and voltage-clamp data were obtained with the brain slice submerged in artificial cerebrospinal fluid. The rats were between 21 and 35 days of age at the time the recordings were made. Synaptic responses were evoked by a bipolar stimulating electrode placed on the lateral lemniscus just ventral to the dorsal nucleus. To eliminate glycinergic inhibitory responses, all physiological data were gathered with 0.5 microM strychnine added to the saline bath. Under current-clamp conditions, excitatory postsynaptic potentials could be subdivided into early and late components. The early component produced a single, highly reliable, short-latency spike and the later component produced a more variable, long-latency spike or train of spikes. The non-N-methyl-D-aspartate antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione, completely blocked the early excitatory postsynaptic potential and its associated action potential. The N-methyl-D-aspartate antagonist, D,L-2-amino-5-phosphonovaleric acid, blocked the later excitatory postsynaptic potential and its action potentials. Typically, both early and late excitatory postsynaptic potentials could be recorded from the same cell, but the early excitatory postsynaptic potential was evoked at lower stimulus levels and had a larger amplitude than the later excitatory postsynaptic potential. Under voltage-clamp conditions, dorsal nucleus of the lateral lemniscus neurons responded to stimulation of the lateral lemniscus with excitatory postsynaptic currents. Outward excitatory postsynaptic currents were recorded with holding potentials that depolarized the cell membrane and inward currents were seen when the cell was hyperpolarized. The current-voltage (I-V) relation of the early peak portion of the excitatory postsynaptic current was nearly linear, whereas the I-V relation of the later excitatory postsynaptic current (12 ms after the peak) was non-linear over the range between -50 and - 100 mV. The outward excitatory postsynaptic current consisted of an early current that was selectively blocked by 6-cyano-7-nitroquinoxaline-2,3-dione and a later current that was blocked by D,L-2-amino-5-phosphonovaleric acid. In artificial cerebrospinal fluid with normal concentrations of Mg2+, the inward excitatory postsynaptic current was blocked by 6-cyano-7-nitroquinoxaline-2,3-dione, but was not affected by D,L-2-amino-5-phosphonovaleric acid. In Mg2+-free artificial cerebrospinal fluid. however, the early component of the inward excitatory postsynaptic current was selectively blocked by 6-cyano-7-nitroquinoxaline-2,3-dione and a later component was blocked by D,L-2-amino-5-phosphonovaleric acid. The results indicate that both N-methyl-D-aspartate and non-N-methyl-D-aspartate receptor-mediated synaptic responses are present in dorsal nucleus of the lateral lemniscus neurons of rats at 21-35 days of age. The N-methyl-D-aspartate component had a longer time-course and a higher threshold than the non-N-methyl-D-aspartate component, and was subject to a voltage-dependent Mg2+ block when the cell's membrane was hyperpolarized. The long-duration N-methyl-D-aspartate component is probably responsible for the prolonged inhibitory effect of dorsal nucleus of the lateral lemniscus neurons on physiological responses in the rat's inferior colliculus.

Factors affecting paired-pulse facilitation in hippocampal CA1 neurons in vitro.

Factors underlying paired-pulse facilitation (PPF) were studied by intracellular and field recordings of CA1 neurons in the hippocampal slice in vitro, following stimulation of the Schaffer collaterals apical dendritic afferents. Similar magnitudes of PPF were found using the slopes or peaks of the excitatory postsynaptic potentials (EPSPs) recorded intracellularly or extracellularly at the soma or dendrites. The paired-pulse EPSP facilitation index (EPI), defined as the ratio of EPSP slope evoked by the second pulse (E2) to that evoked by the first pulse (E1), had a broad peak at 30-60 ms interpulse interval (IPI). EPI was largest at small E1 and decreased with an E1 increase. Spiking excitability was enhanced after the second as compared to the first pulse as evidenced by (1) a decreased latency to fire and (2) an increased tendency to fire double or multiple spikes. The PPF of spiking resulted partly from an increased E2 and partly from a diminished inhibition evoked by the second pulse. Whether the first pulse elicited a spike or not had no significant effect on the EPI. Multiple spiking evoked by the second pulse was partly blocked by the GABAB antagonist CGP35348 (1 mM). The PPF of the EPSP slopes, however, was not significantly affected by GABAB antagonists, GABAA antagonist bicuculline or NMDA antagonist 2-aminophosphonovalerate. In conclusion, PPF may serve as a means of amplification of synaptic transmission such that reliable spike output may result from a given set of synapses.

Analysis of 22 kHz ultrasonic vocalization in laboratory rats: long and short calls.

There is a remarkable variation in the length of single ultrasonic calls emitted by adult rats. The duration of calls is likely to convey information for conspecifics. The goal of the present study was to analyze 22 kHz calls emitted by naive laboratory rats in response to contact with the human hand and to measure their acoustic features, with a particular emphasis on call duration. Repeated hand touch applied to the nape of the neck of rats induced ultrasonic calls, 97.4% of which were within the range of 20-29 kHz and 2.6% of which were within 44-67 kHz. Distribution of duration of 6765 calls revealed two subpopulations of 22 kHz calls: 20-300 ms calls with its peak at 150 ms and calls above 310 ms with highest values at approximately 500-600 ms without a clear peak. These two call populations were referred to as short and long calls, respectively. The short and the long vocalizations contained 80% and 100% of calls within the range of the 22 kHz frequency, respectively. The findings indicated that, in the situation studied, the 22 kHz vocalization of adult rats consists of two distinguishable subpopulation of calls: short and long with the boundary between them at 300 ms.

Glutamate receptors underlying excitatory synaptic transmission in the rat's lateral superior olive studied in vitro.

Glutamate receptors underlying synaptic excitation in the rat's lateral superior olive were studied by whole-cell patch clamp recordings in a brain slice preparation. Recordings from two morphological types of cells, bipolar and multipolar, identified by intracellular labeling with biocytin, showed that there were no obvious differences in responses mediated or modulated by ionotropic and metabotropic receptors between these two types of neurons. The excitatory postsynaptic potentials (EPSPs) elicited by ipsilateral stimulation of the trapezoid body consisted of two components. An earlier component, which had faster rise time constant and decay time constant, was mediated by non-NMDA receptors. A later component, which had slower rise time and decay time constants, was mediated by NMDA receptors. Suprathreshold responses (action potentials), which arose from the early component, were always abolished by the non-NMDA antagonist, CNQX, but not by the NMDA antagonist, APV. These results suggest that both non-NMDA and NMDA receptors are present in LSO neurons, and that fast excitatory transmission in LSO is primarily mediated by non-NMDA receptors. The metabotropic glutamate receptor agonists, t-ACPD and L-AP4, reduced the size of EPSPs evoked by stimulation of the ipsilateral trapezoid body in LSO neurons; the reductive action of t-ACPD was reversed by the antagonist, MCPG, indicating that metabotropic glutamate receptors, probably group II and III subtypes, can modulate excitatory synaptic transmission in LSO.

High-frequency ultrasonic vocalization induced by intracerebral glutamate in rats.

Direct injection of glutamate, a neuroexcitatory agent, into the anterior hypothalamic-preoptic area of the rat brain induced ultrasonic vocalization. This vocalization was characterized by short-duration calls (below 60 ms) of high sound frequency (pitch), mostly above 40 kHz, and was similar to the known 50-kHz vocalization observed in natural situations. The glutamate-induced vocalization was dose dependent within the dose range of 16.9-67.6 micrograms and was antagonized by local pretreatment with MK-801, an NMDA antagonist. The increasing dosage of glutamate induced more calls and had a significant influence on frequency and intensity of emitted ultrasound. The average sound frequency increased whereas the mean sound intensity decreased with the dosage of glutamate. On the other hand, the mean duration of a single call and the bandwidth did not significantly change with doses of glutamate. Injection of carbachol, a muscarinic cholinomimetic agent, into the same brain sites as glutamate, induced a different type of ultrasonic vocalization with low sound frequency and long call duration, known as 22-kHz calls. The results suggest that high sound frequency, short-duration calls (50 kHz) and low sound frequency, long-duration calls (22 kHz) have different neurophysiological and neurochemical mechanisms.

Automated, simplified GC/MS data processing system for organic acidemia screening and its application.

Gas chromatography mass spectrometry (GC/MS) is widely used in diagnosis of organic acidemias. However, GC/MS has not yet become a routine laboratory test, because of the complexity in interpretation of GC/MS data. We developed a personal computer-based system of automated metabolic profiling and disease detection for the screening of organic acidemias by GC/MS. The data were processed after the GC/MS analysis of urinary organic acids. In this system, 130 kinds of metabolites and 25 disorders of organic acids were enrolled for the search and detection, respectively. Metabolites were identified with methylene unit values (MU). target ions (Q- and C-ions) and their intensity ratios, and semiquantified by peak relative area (%) of the Q-ions to that of an internal standard. Metabolites whose values exceeded the cutoff of the control table were flagged as abnormal. The diseases or pathological condition were automatically evaluated by combination of the abnormal compounds. In this system, index metabolites were categorized into three groups. "AND, "OR" and "NO". The groups, "AND" and "OR" comprised essential and optional compounds, respectively, for the specific diagnosis. The third group, "NO", included compounds which must be absent to reach a diagnosis. We compiled data of MU values and mass spectrum of 130 kinds of index metabolites, and tested the usefulness of this system by analysis of 74 patients with 19 kinds of diseases. In all cases, at least a correct diagnosis could be found among the disease names outputted. We have successfully applied this to a pilot neonatal screening by GC/MS in our regional area, and acylglycine analysis by the stable isotope dilution method with tert-butyldimethylsilyl derivatization. With our system, many people can attend for screening programs using GC/MS.

[Cytologic diagnosis of salivary gland lesions by fine needle aspiration].

Fine needle aspiration cytology was used in diagnosis of 504 major and minor salivary gland lesions. In 180 cases with satisfactory specimens, cytologic diagnosis was contrasted with pathohistologic findings. There were 150 benign lesions (including 124 tumors and 26 cases of other lesions) and 30 malignant tumors. The total correspondence rate was 94.5%. The rate of misdiagnosis was 5.5%. Cytologic appearance of various salivary gland tumors is described and the significance of distribution of mucus is discussed. The misdiagnosed cases were mainly in the early stage.

L-carnitine enhances oocyte maturation and development of parthenogenetic embryos in pigs.

The objective was to determine whether adding L-carnitine in IVM/IVC medium enhanced maturation and developmental competence of porcine oocytes in vitro. Oocyte maturation rates did not differ significantly among groups supplemented with 0, 0.25, 0.5, or 1 mg/mL of L-carnitine added during IVM (although 2 mg/mL of L-carnitine reduced maturation rate). Compared with control oocytes, those treated with 0.5 mg/mL of L-carnitine during IVM had greater (P < 0.05) rates of blastocyst formation after parthenogenetic activation, and these blastocysts had less (P < 0.05) apoptosis. Adding 0.5 mg/mL of L-carnitine during IVM also significantly reduced intracellular reactive oxygen species (ROS), and increased glutathione (GSH) concentrations. With or without glucose supplementation, 0.5 mg/mL of L-carnitine in the IVM medium significantly hastened nuclear maturation of oocytes. Moreover, supplementing the IVM medium with either glucose or L-carnitine increased (P < 0.05) percentages of oocytes that reached the metaphase II (MII) stage, relative to a control group. Final maturation rates in IVM medium containing either glucose or L-carnitine were not significantly different. Adding L-carnitine (0 to 2 mg/mL) to IVC medium for activated porcine oocytes did not significantly affect development. However, 0.5 mg/mL of L-carnitine in IVC medium significantly reduced reactive oxygen species levels and apoptosis in activated blastocysts, although glutathione concentrations were not significantly altered. In conclusion, adding L-carnitine during IVM/IVC improved developmental potential of porcine oocytes, and also the quality of parthenogenetic embryos, probably by accelerating nuclear maturation, and preventing oxidative damage and apoptosis.

Diffusion and accumulation in cultivated vegetable plants of di-(2-ethylhexyl) phthalate (DEHP) from a plastic production factory.

Di-(2-ethylhexyl) phthalate (DEHP) concentrations in the atmosphere and in four vegetable crops including Brassica chinensis L. (bok choy), Brassica campestris L. (field mustard), Vigna unguiculata Walp. (cowpea), and Solanum melongena L. (eggplant) cultivated on land surrounding a plastic production factory were determined. The air DEHP concentrations (means) at the sites 0.2, 0.4, 0.8, and 1.6 km away from the plastic production building were about 9.4-12.8, 5.8-9.6, 1.6-5.0, and 0.04-0.27 microg m(-3) dry weight (DW), respectively. Wind direction is a key factor influencing the measurable DEHP concentration of the air, which was highest in the downwind direction and lowest in the upwind direction, and thus the vegetables accumulated the highest DEHP contents in the downwind direction and the lowest quantities in the upwind direction. The highest DEHP accumulations content of bok choy, field mustard, eggplant, and cowpea were 52.0 +/- 3.1, 43.1 +/- 2.2, 36.2 +/- 2.8, and 19.4 +/- 0.47 mg kg(-1) DW, respectively. Safety estimation on the basis of the daily intake limit referenced by the US Environmental Protection Agency (USEPA) led to the conclusion that eating vegetables cultivated 0.2 km away from the plastic production building is not a food safety problem under normal conditions. A strong positive linear correlation between atmospheric DEHP concentration and DEHP content of the vegetable crops was found. The limits for air DEHP concentration for safe vegetable cultivation are 24.0, 34.8, 40.8, and 82.8 microg m(-3) for bok choy, field mustard, cowpea, and eggplant, respectively, by calculating from the equation of linear regression between air DEHP concentration and vegetable DEHP content.