Effects of social context manipulation on dorsal and ventral hippocampal neuronal responses.

The hippocampus is critical for contextual memory and has recently been implicated in various kinds of social memory. Traditionally, studies of hippocampal context coding have manipulated elements of the background environment, such as the shape and color of the apparatus. These manipulations produce large shifts in the spatial firing patterns, a phenomenon known as remapping. These findings suggest that the hippocampus encodes and differentiates contexts by generating unique spatial firing patterns for each environment a subject encounters. However, we do not know whether the hippocampus encodes social contexts defined by the presence of particular conspecifics. We examined this by exposing rats to a series of manipulations of the social context, including the presence of familiar male, unfamiliar male and female conspecifics, in order to determine whether remapping is a plausible mechanism for encoding socially-defined contexts. Because the dorsal and ventral regions of the hippocampus are thought to play different roles in spatial and social cognition, we recorded neurons in both regions. Surprisingly, we found little evidence of remapping in response to manipulation of the social context in either the dorsal or ventral hippocampus, although we saw typical remapping in response to changing the background color. This result suggests that remapping is not the primary mechanism for encoding different social contexts. However, we found that a subset of hippocampal neurons fired selectively near the cages that contained the conspecifics, and these responses were most prevalent in the ventral hippocampus. We also found a striking increase in the spatial information content of ventral hippocampal firing patterns. These results indicate that the ventral hippocampus is sensitive to changes in the social context and neurons that respond selectively near the conspecific cages could play an important, if not fully understood role in encoding the conjunction of conspecifics, their location and the environment.

Affective mirror and anti-mirror neurons relate to prosocial help in rats.

Although empathic emotion is closely related to prosocial behavior, neuronal substrate that accounts for empathy-associated prosocial action remains poorly understood. We recorded neurons in the anterior cingulate cortex (ACC) and insular cortex (InC) in rats when they observed another rat in pain. We discovered neurons with anti-mirror properties in the ACC and InC, in addition to those with mirror properties. ACC neurons show higher coupling between activation of self-in-pain and others-in-pain, whereas the InC has a higher ratio of neurons with anti-mirror properties. During others-in-pain, ACC neurons activated more when actively nose-poking toward others and InC neurons activated more when freezing. To further illustrate prosocial function, we examined neuronal activities in the helping behavior test. Both ACC and InC neurons showed specific activation to rat rescuing which is contributed by mirror, but not anti-mirror neurons. Our work indicates the functional involvement of mirror neuron system in prosocial behaviors.

Effect of low-level creatinine clearance on short-term postoperative complications in patients with colorectal cancer.

Background: Renal function is closely related to cancer prognosis. Since preoperative renal insufficiency has been identified as a risk factor for postoperative complications, this study aimed to investigate the effect of preoperative creatinine clearance rate (CrCl) on short-term prognosis of patients undergoing colorectal surgery. Methods: A retrospective analysis was conducted of the electronic health records of 526 adult patients who underwent elective colorectal cancer (CRC) surgery from September 2014 to February 2019 at the First Affiliated Hospital of Wenzhou Medical University. Cases were divided into two groups according to CrCl level and clinical variables were compared. Risk factors associated with postoperative complications were evaluated through univariate and multivariate logistic regression analyses. Results: A total of 526 patients met the inclusion criteria. The overall rate of postoperative complications was 28.14%. Overall, the incidence of postoperative complications was significantly higher in the low CrCl patients. A low-level CrCl, multi-organ combined resection, and Charlson comorbidity index (CCI) were independent risk factors for short-term complications in patients with CRC. However, a low CrCl was identified as an independent risk factor for short-term postoperative complications in elderly, but not young patients in a subgroup analysis. Conclusions: Preoperative low-level CrCl, multi-organ combined resection, and CCI were significant risk factors of postoperative complications in CRC patients. Preoperative low-level CrCl and multi-organ combined resection has a poor prognostic impact for elderly patients with CRC. These findings should have important implications for health care decision-making among patients with CRC who are at higher risk for post-operative complications.

Effects of temperature and salinity on antioxidant responses in livers of temperate (Dicentrarchus labrax) and tropical (Chanos Chanos) marine euryhaline fish.

Temperature and salinity are abiotic factors that affect physiological responses in aquaculture species. The European sea bass (Dicentrarchus labrax) is a temperate species that is generally farmed at 18 °C in seawater (SW). In the wild, its incursions in shallow habitats such as lagoons may result in hyperthermal damage despite its high thermal tolerance. Meanwhile, the milkfish (Chanos chanos), a tropical species, is generally reared at 28 °C, and in winter, high mortality usually occurs under hypothermal stress such as cold snaps. This study compared changes in hepatic antioxidant enzymes (superoxide dismutase, SOD; and catalase, CAT) in these two important marine euryhaline aquaculture species in Europe and Southeast Asia, respectively, under temperature challenge combined with hypo-osmotic (fresh water, FW) stress. After a four-week hyper- or hypo-thermal treatment, hepatic SOD activity was upregulated in both species reared in SW and FW, indicating enhanced oxidative stress in European sea bass and milkfish. The expression profiles of sod isoforms suggested that in milkfish, the increase in reactive oxygen species (ROS) was mainly at the cytosol level, leading to increased sod1 expression. In European sea bass, however, no obvious difference was found between the expression of sod isoforms at different temperatures. A lower expression of sod2 was observed in FW compared to SW in the latter species. Moreover, no significant change was observed in the mRNA expression and activity of CAT in the livers of these two species under the different temperature treatments, with the exception of the lower CAT activity in milkfish challenged with SW at 18 °C. Taken together, our results indicated that the antioxidant responses were not changed under long-term hypoosmotic challenge but were enhanced during the four-week temperature treatments in livers of both the temperate and tropical euryhaline species.

AWGS2019 vs EWGSOP2 for diagnosing sarcopenia to predict long-term prognosis in Chinese patients with gastric cancer after radical gastrectomy.

BACKGROUND: Sarcopenia is a nutrition-related disease and has a profound effect on the long-term overall survival (OS) of patients with gastric cancer. Its diagnostic criterion is critical to clinical diagnosis and treatment. However, previous research reported widely differing sarcopenia prevalence due to different criteria. AWGS2019 and EWGSOP2 are the two latest and widely adopted criteria. AIM: To compare the effects of AWGS2019 and EWGSOP2 on the long-term OS of Chinese gastric cancer patient after radical gastrectomy. METHODS: An observational study was conducted from July 2014 to January 2017, which included 648 consecutive gastric cancer patients who underwent radical gastrectomy. The sarcopenia elements (skeletal muscle index, handgrip strength, and gait speed) were measured within 1 mo or 7 d before surgery. The patients were followed at fixed intervals to gain the outcomes. Multivariate Cox regression analysis was performed to determine the association between sarcopenia and the long-term OS of these patients according to the two criteria separately. The predictive performance of the models with AWGS2019 and EWGSOP2 were evaluated by the concordance index (C-index) and area under the time-dependent receiver operating characteristic curve (AUC). The Akaike information criterion (AIC) was applied to compare model fits. RESULTS: The prevalence of sarcopenia was 20.5% and 11.3% according to AWGS2019 and EWGSOP2, respectively. Sarcopenia was an independent risk factor for the long-term OS no matter based on AWGS2019 or EWGSOP2, but AWGS2019-sarcopenia in multivariate model had a higher hazard ratio (HR) [2.150 (1.547-2.988)] than EWGSOP2-sarcopenia [HR 1.599 (1.092-2.339)]. Meanwhile, the model with AWGS2019-sarcopenia [C-index 0.773 (0.742-0.804); AIC 2193.7; time-dependent AUC 0.812 (0.756-0.867) for 1-year OS, 0.815 (0.778-0.852) for 3-year OS, and 0.809 (0.759-0.859) for 5-year OS] had better predictive power and model fits than the model with EWGSOP2-sarcopenia [C-index 0.762 (0.729-0.795); AIC 2215.2; time-dependent AUC 0.797 (0.741-0.854) for 1-year OS, 0.804 (0.767-0.842) for 3-year OS, and 0.799 (0.748-0.850) for 5-year OS]. CONCLUSION: Sarcopenia is an independent risk factor for the long-term OS in Chinese gastric cancer patients undergoing radical gastrectomy. The prediction model with AWGS2019-sarcopenia has better predictive power and model fits than the prediction model with EWGSOP2-sarcopenia. AWGS2019 may be more appropriate for diagnosing sarcopenia in these Chinese patients than EWGSOP2.

Synthesis, anticancer evaluation and mechanism studies of novel indolequinone derivatives of ursolic acid.

A series of novel indolequinone derivatives of ursolic acid bearing ester, hydrazide, or amide moieties were designed, synthesized, and screened for their in vitro antiproliferative activities against three cancer cell lines (MCF-7, HeLa, and HepG2) and a normal gastric mucosal cell line (Ges-1). A number of compounds showed significant activity against tested cancer cell lines. Among them, compound 6t exhibited the most potent activity against three cancer cell lines with IC50 values of 1.66 ± 0.21, 3.16 ± 0.24, and 10.35 ± 1.63 µM, respectively, and considerably lower cytotoxicity to Ges-1 cells. Especially, compound 6t could arrest cell cycle at S phase, suppress the migration of MCF-7 cells, elevate intracellular reactive oxygen species (ROS) level, and decrease mitochondrial membrane potential. Western blot analysis showed that compound 6t upregulated Bax, cleaved caspase-3/9, cleaved PARP levels and downregulated Bcl-2 level of MCF-7 cells. All these results indicated that compound 6t could significantly induce the apoptosis of MCF-7 cells. Meanwhile, compound 6t markedly decreased p-AKT and p-mTOR expression, which revealed that compound 6t probably exerted its cytotoxicity through targeting PI3K/AKT/mTOR signaling pathway. Therefore, compound 6t could be a promising lead for the discovery of novel anticancer agents.

Energy response and fatty acid metabolism in Onychostoma macrolepis exposed to low-temperature stress.

Temperature is a key environmental factor, and understanding how its fluctuations affect physiological and metabolic processes is critical for fish. The present study characterizes the energy response and fatty acid metabolism in Onychostoma macrolepis exposed to low temperature (10 °C). The results demonstrated that cold stress remarkably disrupted the energy homeostasis of O. macrolepis, then the AMP-activated protein kinase (AMPK) could strategically mobilize carbohydrates and lipids. In particular, when the O. macrolepis were faced with cold stress, the lipolysis was stimulated along with the enhanced fatty acid ß-oxidation for energy, while the fatty acid synthesis was supressed in the early stage. Additionally, the fatty acid composition analysis suggested that saturated fatty acid (SFA) might accumulate while monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) in storage lipids (mainly containing non-polar lipid, NPL) could be utilized to supply energy during cold acclimation. Altogether, this study may provide some meritorious for understanding the cold-tolerant mechanism of fish in the viewpoint of energy balance combined with fatty acid metabolism, and thus to contribute to this species rearing in fish farms in the future.

Potential osmoprotective roles of branchial heat shock proteins towards Na+, K+-ATPase in milkfish (Chanos chanos) exposed to hypotonic stress.

In euryhaline teleosts, osmoregulatory mechanisms vary with osmotic stresses, and heat shock proteins (HSPs) play a central role in maintaining cellular homeostasis. The present study aimed to investigate the expression and potential roles of HSP70 and HSP90 in the gills of seawater (SW)- and freshwater (FW)-acclimated milkfish (Chanos chanos). Four HSP genes, including Cchsc70 (heat shock cognate 70), Cchsp70, Cchsp90α, and Cchsp90ß, were analyzed in milkfish gills. Among these genes, only the mRNA abundance of branchial Cchsp90α was significantly lower in the FW-acclimated than in the SW-acclimated milkfish. Immunoblotting showed no significant difference in the relative protein abundance of branchial HSP70 and HSP90 between the two groups. The time-course experiments (from SW to FW) showed that the protein abundance of HSP70 and HSP90 at the 3 h and 6 h post-transfer and then declined gradually. To further illustrate the potential osmoregulatory roles of HSP70 and HSP90, their interaction with Na+, K+-ATPase (NKA, the primary driving force for osmoregulation) was analyzed using co-immunoprecipitation. The results showed the interaction between HSP70, HSP90 and NKA after acclimation to SW or FW increased within 3 h; and then returned to normal levels within 7 days. To our knowledge, the present study was the first to demonstrate that the interaction between HSP70, HSP90 and NKA changes with hypotonic stress in euryhaline teleosts. Before the transfer, no interaction was detected. When transferred to FW from SW, the interaction of HSP70 and HSP90 with NKA were detected. The results suggested that HSP70 and HSP90 participated in the acute responses of osmoregulatory mechanisms to protect branchial NKA from hypotonic stress in milkfish.

Positive correlation of gene expression between branchial FXYD proteins and Na+/K+-ATPase of euryhaline milkfish in response to hypoosmotic challenges.

FXYD proteins are crucial regulators of Na+/K+-ATPase (NKA), which plays an important role in ion exchange by providing the driving force for other ion-transporting systems in the osmoregulatory organs, including the gills. In milkfish (Chanos chanos), gill NKA has been widely investigated and found to alter its expression (both mRNA and protein) and activity in response to environmental salinity changes. However, the expression and roles of the regulatory proteins of NKA, the FXYD proteins, in milkfish gills upon salinity challenge is not yet clear. Hence, this study illustrated the potential roles of milkfish branchial FXYD proteins in modulating NKA expression via identification and tissue distributions of FXYD proteins, as well as the effects of salinity on expression of gill fxyd and nka mRNA. Six milkfish FXYD proteins (CcFXYD) were identified. In milkfish gill, gill-specific Ccfxyd11 was the predominant member, followed by Ccfxyd9 and Ccfxyd8. Upon hypoosmotic challenges, increases in gill Ccfxyd11, Ccfxyd8, Ccnka α1, and Ccnka ß1 mRNA as well as significantly positive correlations were observed. Moreover, after acute salinity changes, expression of gill Ccfxyd11 and Ccnka was found to change with ambient salinity, and significant positive correlations were also exhibited between Ccfxyd11 and Ccnka α1. Overall, these results revealed close relationships between CcFXYD11 and CcNKA α1 in milkfish gills, highlighting the potential roles of CcFXYD11 in osmoregulation.

Nocifensive behavior-related laser heat-evoked component in the rostral agranular insular cortex revealed using morphine analgesia.

The rostral agranular insular cortex (RAIC), an opioid-responsive site, is essential for modulating nociception in rats. Our previous studies have shown that morphine suppressed long latency laser heat-evoked nociceptive responses in the primary somatosensory cortex (SmI). By contrast, morphine significantly attenuated both short and long latency responses in the anterior cingulate cortex (ACC). The present study assessed the effect of morphine on laser heat-evoked responses in the RAIC. Laser heat irradiation applied to the rat forepaws at graded levels was used as a specific noxious stimulus. In the RAIC, the first part of the long latency component (140-250ms) of the laser heat-evoked response was enhanced by intraperitoneal morphine (5mg/kg). When the laser heat-evoked cortical responses were examined for trials showing strong nocifensive movement (paw licking), moderate nocifensive movement (paw lifting), and no nocifensive movement, a 140-250ms period enhancement was observed in the RAIC only for the paw lifting movement. This enhancement was absent in the SmI. Thus, our data suggest that the RAIC has a pain-related behavior-dependent neuronal component. Furthermore, the RAIC, ACC, and SmI are differentially modulated by morphine analgesia.

[Expression and significance of mRNA and protein of eIF4E, p-eIF4E and MCl-1 in pathological scar].

OBJECTIVE: To study the expression of eIF4E, p-eIF4E (Ser 209) and Mcl-1 gene in the pathological scars and to investigate its role and its probable mechanism in the pathogenesis of abnormal scar. METHODS: Quantitative real-time PCR and Western Blot was performed to detect the expression and distribution of mRNA and protein of eIF4E and Mcl-1 in hypertrophic scar (10 cases), keloid (10 cases), normal scar (10 cases), and normal skin (10 cases). Western Blot was performed to detect the expression and distribution of protein of p-eIF4E in hypertrophic scar (10 cases), keloid (10 cases), normal scar (10 cases), and normal skin (10 cases). RESULTS: The expression of eIF4E mRNA and protein were 1.38 +/- 0.45, 1.23 +/- 0.23 in the normal skin (10 cases); 5.400 +/- 0.450, 5.460 +/- 0.460 in normal scar (10 cases); 0.597 +/- 0.060, 0.590 +/- 0.040 in hypertrophic scar (10 cases) and 0.694 +/- 0.066, 0.697 +/- 0.022 in keloid (10 cases). The expression of p-eIF4E protein in the normal skin (10 cases), normal scar (10 cases), hypertrophic scar (10 cases), and keloid (10 cases) were 0.202 +/- 0.037, 0.216 +/- 0.019, 0.426 +/- 0.026, 0.433 +/- 0.027. The expression of Mcl-1 mRNA and protein were 1.510 +/- 0.660, 1.400 +/- 0.530 in the normal skin (10 cases); 6.65 +/- 0.85, 7.23 +/- 1.53 in normal scar (10 cases); 0.589 +/- 0.059, 0.660 +/- 0.063 in hypertrophic scar (10 cases) and 0.870 +/- 0.118, 0.914 +/- 0.064 in the keloid (10 cases). The positive rate of mRNA and protein of eIF4E and Mcl-1 was not statistically different between the hypertrophic scar and keloid (P > 0.05), while they were all remarkably significant between normal scar and abnormal scar (P < 0.05). The phosphorylation of eIF4E in pathological scar was higher than that in control group. In pathological scar, mRNA and protein of eIF4E and Mcl-1 showed a strong positive correlation. CONCLUSIONS: The result indicates that the expression of eIF4E, p-eIF4E and Mcl-1 is increased in pathological scar. eIF4E plays an important role in pathological scar. Its activity is regulated by its phosphorylation. Therefore, eIF4E, p-eIF4E and Mcl-1 overexpression may play an important role in the proliferation of fibroblasts and in the pathogenesis of pathological scar.

Sleep disturbance among spontaneously hypertensive rats is mediated by an α1-adrenergic mechanism.

BACKGROUND: Inadequate sleep may aggravate hypertension, but the pathophysiology of sleep disturbance in hypertension remains unknown. Among spontaneously hypertensive rats (SHR), sleep disturbance co-occurred with sympathetic hyperactivity; therefore, we hypothesized that the sleep disturbance can be alleviated by antagonizing the adrenergic overdrive. METHODS: Polysomnographic recording was performed in SHR by telemetry. The animals were first injected with saline, and 2 days later with a hypotensive agent. Cardiac and vascular sympathetic activity were assessed using the normalized low-frequency power (LF%) of heart rate variability and the low-frequency power of arterial pressure variability (BLF), respectively. RESULTS: A comparison was made between the saline and hypotensive drug treatments. During quiet sleep (QS), the α1-blocker prazosin induced a significant decrease in BLF, but had no effect on LF%. The total time and bout duration of QS were lengthened and QS interruption was reduced (P < 0.05 for all). When both α1- and α2-adrenoceptors were blocked by phentolamine, both BLF and LF% were lower (P < 0.05 for both), but no modification to sleep structure could be observed. To antagonize ß-adrenergic activity, atenolol and propranolol were injected. The LF% after either antagonist treatment was significantly decreased; however, sleep structure was not significantly changed. The QS-promoting effect of prazosin is specific to SHR, because prazosin is ineffective when administered to Wistar-Kyoto rats. CONCLUSIONS: α1-adrenergic antagonism may reverse, at least partially, the poor sleep quality of SHR, suggesting a vicious cycle can be established between adrenergic overdrive and sleep disturbance.

Potentially predictive microRNAs of gastric cancer with metastasis to lymph node.

AIM: To detect the expression of 60 microRNAs (miRNAs) in gastric cancer tissues and find new predictive biomarkers of gastric cancer with metastasis. METHODS: The expressions of 60 candidate miRNAs in 30 gastric cancer tissues and paired normal tissues were detected by stem-loop real-time reverse transcription-polymerase chain reaction. After primary screening of miRNAs expression, 5 selected miRNAs were further testified in another 22 paired gastric tissues. Based on the expression level of miRNAs and the status of metastasis to lymph node (LN), receiver-operating-characteristic (ROC) curve were used to evaluate their ability in predicting the status of metastasis to LN. RESULTS: Thirty-eight miRNAs expressions in gastric cancer tissues were significantly different from those in paired normal tissues (P < 0.01). Among them, 31 miRNAs were found to be up-expressed in cancer tissues and 1 miRNAs were down-expressed ≥ 1.5 fold vs paired normal gastric tissue. Five microRNAs (miR-125a-3p, miR-133b, miR-143, miR-195 and miR-212) were differently expressed between different metastatic groups in 30 gastric cancer biopsies (P < 0.05). Partial correlation analysis showed that hsa-mir-212 and hsa-mir-195 were correlated with the status of metastasis to LN in spite of age, gender, tumor location, tumor size, depth of invasion and cell differentiation. ROC analysis indicated that miR-212 and miR-195 have better sensitivities (84.6% and 69.2%, respectively) and specificities (both 100%) in distinguishing biopsies with metastasis to LN from biopsies without metastasis to LN. CONCLUSION: miR-212 and miR-195 could be independent biomarkers in predicting the gastric cancer with metastasis to LN.

Transvaginal cystocele repair using pursestring technique reinforced with custom-tailored two-armed mesh.

OBJECTIVE: To evaluate the efficacy and safety of vaginal cystocele repair using a pursestring suture technique reinforced with custom-tailored 2-armed mesh. METHODS: The records of patients who underwent isolated cystocele repair by a single surgeon were retrospectively reviewed. All women were assessed by clinical examination and lower urinary tract symptoms preoperatively and at 3 and 6 months and every year postoperatively. The principal outcome measure was anatomic cure (stage 1 or lower) assessed by the Pelvic Organ Prolapse Quantification system. RESULTS: From May 2005 to June 2009, 50 consecutive patients with minimum stage 2 (Aa or Ba 0) cystocele were treated using the procedure. The mean follow-up was 24 months (range 12-54), and the success rate was 96% (48 of 50). No intraoperative complications occurred. Two developed vaginal erosion (4%) and underwent excision of the extruded mesh smoothly. The final 21 consecutive patients underwent urodynamic assessment pre- and postoperatively. Compared with the baseline urodynamic findings, surgery resulted in decreased maximal urethral pressure and maximal urethral closure pressures. However, neither increased de novo incontinence nor reduced cystometric bladder capacity developed. CONCLUSION: The use of the pursestring suture technique reinforced with custom-tailored 2-armed mesh for vaginal cystocele repair is safe, effective, and economic.

[Expression and significance of P57(kip2) and Maspin in pathological scar].

OBJECTIVE: To study the expression of P57(kip2) and Maspin in the pathological scar and their possible role in the pathogenesis of abnormal scars. METHODS: Immunohistochemistry integrated image analysis and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of P57(kip2) and Maspin in hypertrophic scar, keloid, mature scar and normal skin. Statistics was used to analyze the datas. RESULTS: The expression of P57(kip2) protein was fixed to fibroblast intranuclear in abnormal scar, and the expression of P57(kip2) protein and P57(kip2) mRNA decreased (P < 0.05). The expression of Maspin protein was fixed to fibroblast cytoplasm and intranuclear in abnormal scar, and the expression of Maspin protein and Maspin mRNA decrease, compared with that in normal group (P < 0.05). There was positive correlation between P57(kip2) protein and Maspin protein expression (P < 0.01). CONCLUSIONS: The decreased expression of P57(kip2) and Maspin in abnormal scar shows that they are cicatrix-related genes. There is a positive relationship between the two genes. It may be one of the mechanisms of pathogenesis of abnormal scar. It makes effect through fibroblasts.

Elevated Na+/K+-ATPase responses and its potential role in triggering ion reabsorption in kidneys for homeostasis of marine euryhaline milkfish (Chanos chanos) when acclimated to hypotonic fresh water.

The milkfish (Chanos chanos) is an economic species in Southeast Asia. In Taiwan, the milkfish are commercially cultured in environments of various salinities. Na(+)/K(+)-ATPase (NKA) is a key enzyme for fish iono- and osmoregulation. When compared with gills, NKA and its potential role were less examined by different approaches in the other osmoregulatory organs (e.g., kidney) of euryhaline teleosts. The objective of this study was to investigate the correlation between osmoregulatory plasticity and renal NKA in this euryhaline species. Muscle water contents (MWC), plasma, and urine osmolality, kidney histology, as well as distribution, expression (mRNA and protein), and specific activity of renal NKA were examined in juvenile milkfish acclimated to fresh water (FW), seawater (SW 35 per thousand), and hypersaline water (HSW 60 per thousand) for at least two weeks before experiments. MWC showed no significant difference among all groups. Plasma osmolality was maintained within the range of physiological homeostasis in milkfish acclimated to different salinities, while, urine osmolality of FW-acclimated fish was evidently lower than SW- and HSW-acclimated individuals. The renal tubules were identified by staining with periodic acid Schiff's reagent and hematoxylin. Moreover, immunohistochemical staining showed that NKA was distributed in the epithelial cells of proximal tubules, distal tubules, and collecting tubules, but not in glomeruli, of milkfish exposed to different ambient salinities. The highest abundance of relative NKA alpha subunit mRNA was found in FW-acclimated milkfish rather than SW- and HSW-acclimated individuals. Furthermore, relative protein amounts of renal NKA alpha and beta subunits as well as NKA-specific activity were also found to be higher in the FW group than SW and the HSW groups. This study integrated diverse levels (i.e., histological distribution, gene, protein, and specific activity) of renal NKA expression and illustrated the potential role of NKA in triggering ion reabsorption in kidneys of the marine euryhaline milkfish when acclimated to a hypotonic FW environment.

[Correction of severe depression deformities at lower eyelid].

OBJECTIVE: To evaluate the application of temporal fascia flaps in the correction of severe depression deformities at lower eyelids. METHODS: Severe depression deformities at lower eyelid were corrected with temporal fascia flaps pedicled with superficial temporal artery in 9 cases. RESULTS: All flaps survived with good appearance. All patients were followed up for 6-12 months with good long-term results. The donor sites had no obvious scalp scar. CONCLUSIONS: Temporal fascia flap is an optimal choice for correction of the severe depression deformities at lower eyelid. It is easily performed with good result and less donor site morbidity.

[Island skin flap supplied by superficial temporal vessel for the whole ear lobe defect].

OBJECTIVE: To introduce a new method for reconstruction of the whole ear lobe defect. METHODS: The free island skin flap supplied by superficial temporal vessel which was designed at the area anterior and superior to crus helicis. The flap was transferred through subcutaneous tunnel and self-folded to reconstruct the whole ear lobe defect. RESULTS: Since 1999, 6 cases were treated with no complication. The ear lobe shape and skin colour were very natural. CONCLUSIONS: The island skin flap supplied by superficial temporal vessel is very suitable for the whole ear lobe defect with good cosmetic results.

Simultaneous determination of cefepime and vancomycin in plasma and cerebrospinal fluid by MEKC with direct sample injection and application for bacterial meningitis.

A simple MEKC with UV detection at 214 nm for simultaneous analysis of cefepime and vancomycin in plasma and in cerebrospinal fluid (CSF) by direct injection without any sample pretreatment is described. The separation of cefepime and vancomycin from biological matrices was performed at 25 degrees C using a BGE consisting of a Tris buffer with SDS and methanol as the electrolyte solution. Under optimal MEKC conditions for biological samples, good separations with high efficiency and short analysis time are achieved. Several parameters affecting the separation of the drugs from biological matrices were studied, including methanol, pH, and concentrations of the Tris buffer and SDS. The linear ranges of the method for the determination of cefepime and vancomycin in plasma and in CSF using imidazole or cefazolin as an internal standard, respectively, were all over the range of 1-30 microg/mL; the detection limits of cefepime and vancomycin in biological matrices (injection 10 kV, 15 s) were 0.3 and 0.5 microg/mL, respectively. The applicability of the proposed method for the determination of cefepime and vancomycin in plasma and CSF collected after intravenous administration of the drugs in patients with meningitis was demonstrated.