2020年 · 第133卷第15期

During the past decade, huge advantages have been made in mapping and understanding the human-associated intestinal microbiome and its relation with human health. The vast amount of microorganisms that reside in our gastrointestinal tract is being acknowledged for its role in human physiology and due to this role, it has even been referred to as an additional organ.^[1]As a result of the revelation of links between diseases and changes in the intestinal bacterial composition, medical interest in the deliberate use of bacteria or their products in treating diseases, called bacteriotherapy, has mounted likewise. Currently, bacteriotherapy comprises (fecal) microbial transfer, where stools or microbial stool-derivates like washed microbiota^[2]from a donor are administered to treat an illness in an acceptor on the one hand, and probiotics on the other. According to the definition, probiotics are "live microorganisms which when administered in adequate amounts confer a health benefit on the host."^[3]These benefits occur through interaction with the (local) immune system, through the production of desired metabolites and cross-feeding with present microorganisms but are also linked with increasing the bacterial load. Many different bacterial and yeast strains have been studied for their probiotic effects. Probiotics are generally administered in doses of up to 10¹⁰viable microorganisms which are added to the estimated 10¹⁴bacteria in the human gastrointestinal tract.^[4]Upon administration, probiotics are considered to exert a transient effect, without permanent colonization of the host after the treatment has been ceased. In general, wash-out periods of 2 to 4 weeks are being used in clinical trials. Although the term 'probiotics’ is highly useful to describe this type of microorganism based therapy, it also leads to misconceptions. After all, the term probiotic covers many different microorganisms that have very different properties. Taxonomically, probiotics cross even different domains as both unicellular prokaryotic (bacteria) and complex cell eukaryotic (yeast) probiotic strains are being used. Moreover, health benefits of probiotics have been described for different indications and their mode of action is divers. As a result, caution is needed when summarizing the published research results to evaluate the efficacy of probiotics. If we want to improve the potency of probiotic use, there is an urgent need to also recognize the microbial view-point. Here, probiotics are approached from a microbiologist perspective and pinpointed to bacterial probiotics for intestinal applications. The focus lays on the impact of specific supplementation of bacterial strains to an existing microbial ecosystem, starting with the probiotic strain itself.

Hypothermia is associated with many adverse clinical outcomes in pediatric patients, and thus, it is important to find an effective and safe method for preventing peri-operative hypothermia and its associated adverse outcomes in pediatric patients. This study aimed to investigate the effect of forced-air warming blankets with different temperatures on changes in the transforming growth factor-β (TGF-β), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-10 levels in children undergoing surgical treatment for developmental displacement of the hip (DDH).

The study included 123 children undergoing surgery for DDH under general anesthesia. The patients were randomly assigned to three groups, using a random number table: the 32, 38, and 43°C groups according to the temperature setting of the forced-air warming blankets. For each patient, body temperature was recorded immediately after anesthesia induction and intubation (T₀), at initial incision (T₁), at 1 h after incision (T₂), at 2 h after incision (T₃), at the end of surgery (T₄), immediately upon return to the ward after surgery (T₅), and then at 12 h (T₆), 24 h (T₇), 36 h (T₈), and 48 h (T₉) after the surgery. The serum levels of TGF-β, TNF-α, IL-1β, and IL-10 were measured at T₀ and T₄ for all groups.

The number of patients with fever in the 38°C group was significantly less than those in the 32 and 43°C groups (χ² = 6.630, P= 0.036). At T₀, the body temperatures in the 38 and 43°C groups were significantly higher than that in the 32°C group (F = 17.992, P < 0.001). At T₂, the body temperature was significantly higher in the 43°C group than those in the 32 and 38°C groups (F= 12.776, P < 0.001). Moreover, at T₄, the serum levels of TGF-β (F = 3286.548, P < 0.001) and IL-10 (F = 4628.983, P < 0.001) were significantly increased in the 38°C group, and the serum levels of TNF-α (F= 911.415, P < 0.001) and IL-1β (F = 322.191, P < 0.001) were significantly decreased in the 38°C group, compared with the levels in the 32 and 43°C groups.

Force-air warming blankets set at 38°C maintained stable body temperature with less adverse outcome and effectively inhibited the inflammatory response in pediatric patients undergoing surgery for DDH.

ChiCTR1800014820; http://www.chictr.org.cn/showproj.aspx?proj=25240.

Non-coding RNAs have attracted considerable attention for their vital role in cancer. The purpose of this study was to determine the effects of non-coding RNAs on hepatocellular carcinoma (HCC) and reveal their regulatory mechanism in the pathophysiological process.

We measured the expression of mucin 1 (MUC1) and miR-485-5p in tissues from 15 HCC patients and in liver cancer cell lines by quantitative real-time polymerase chain reaction and Western blot, screened for aberrantly expressed microRNAs (miRNAs) by miRNA microarrays. Bioinformatics tools were used to find the miRNA and circular RNA that regulated MUC1, which were validated by RNA immunoprecipitation assay and luciferase reporter assay. Cell counting kit-8, Transwell assays, and flow cytometry were used to conduct functional experiments. Proteins were examined by western blot and immunohistochemical staining assays. Significant differences between groups were estimated using the one-way analysis of variance. A P < 0.05 was considered statistically significant.

MUC1 was overexpressed in HCC tissues compared with that in paratumor tissues (normal vs. tumor, 1.007 ± 0.215 vs. 75.213 ± 18.403, t = 18.401, P < 0.001) while miR-485-5p was down-regulated (normal vs. tumor, 4.894 ± 0.684 vs. 1.586 ± 0.398, t= 16.191, P < 0.001). Inhibition of miR-485-5p promoted cell proliferation (73.33% ± 5.13% vs. 41.33% ± 3.51%, t= 8.913, P < 0.001), migration (102 ± 8 cells vs. 46 ± 8 cells, t= 8.681, P < 0.001), invasion (59 ± 7 cells vs. 28 ± 2 cells, t = 8.034, P < 0.01), and suppressed apoptosis (22.64% ± 6.97% vs. 36.33% ± 3.96%, t = 2.958, P < 0.05) of HepG2 cells with which MUC1 is knocked down. Mechanically, miR-485-5p binds to MUC1, while circHECTD1 binds to miR-485-5p, resulting in the indirect up-regulation of the MUC1 level.

Our findings reveal that circHECTD1 facilitates HCC progression by sponging miR-485-5p to up-regulate MUC1.

Cardiac remodeling after acute myocardial infarction (AMI) is an important process. The present study aimed to assess the protective effects of astaxanthin (ASX) on cardiac remodeling after AMI.

The study was conducted between April and September 2018. To create a rat AMI model, rats were anesthetized, and the left anterior descending coronary artery was ligated. The rats in the ASX group received 10 mg·kg^-1·day^-1 ASX by gavage for 28 days. On the 1st day after AMI, but before ASX administration, six rats from each group were sacrificed to evaluate changes in the heart function and peripheral blood (PB) levels of inflammatory factors. On the 7th day after AMI, eight rats from each group were sacrificed to evaluate the PB levels of inflammatory factors and the M2 macrophage count using both immunofluorescence (IF) and flow cytometry (FC). The remaining rats were observed for 28 days. Cardiac function was examined using echocardiography. The inflammatory factors, namely, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-10, were assessed using enzyme-linked immunosorbent assay. The heart weight/body weight (BW), and lung weight (LW)/BW ratios were calculated, and myocardial fibrosis in the form of collagen volume fraction was measured using Masson trichrome staining. Hematoxylin and eosin (H&E) staining was used to determine the myocardial infarct size (MIS), and TdT-mediated dUTP nick-end labeling staining was used to analyze the myocardial apoptosis index. The levels of apoptosis-related protein, type I/III collagen, transforming growth factor β1 (TGF-β1), metalloproteinase 9 (MMP9), and caspase 3 were assessed by Western blotting. Unpaired t-test, one-way analysis of variance, and non-parametric Mann-Whitney test were used to analyze the data.

On day 1, cardiac function was worse in the ASX group than in the sham group (left ventricular end-systolic diameter [LVID_s]: 0.72 ± 0.08 vs. 0.22 ± 0.06 cm, t= -11.38; left ventricular end-diastolic diameter [LVID_d]: 0.89 ± 0.09 vs. 0.48 ± 0.05 cm, t= -9.42; end-systolic volume [ESV]: 0.80 [0.62, 0.94] vs. 0.04 [0.03, 0.05] mL, Z = -2.89; end-diastolic volume [EDV]: 1.39 [1.03, 1.49] vs. 0.28 [0.22, 0.32] mL, Z = -2.88; ejection fraction [EF]: 0.40 ± 0.04 vs. 0.86 ± 0.05, t= 10.00; left ventricular fractional shortening [FS] rate: 0.19 [0.18, 0.20] %FS vs. 0.51 [0.44, 0.58] %FS, Z = -2.88, all P < 0.01; n = 6). The levels of inflammatory factors significantly increased (TNF-α: 197.60 [133.89, 237.94] vs. 50.48 [47.21 57.10] pg/mL, Z= -2.88; IL-1β: 175.23 [160.74, 215.09] vs. 17.78 [16.83, 19.56] pg/mL, Z = -2.88; IL-10: 67.64 [58.90, 71.46] vs. 12.33 [11.64, 13.98] pg/mL, Z = -2.88, all P < 0.01; n = 6). On day 7, the levels of TNF-α and IL-1β were markedly lower in the ASX group than in the AMI group (TNF-α: 71.70 [68.60, 76.00] vs. 118.07 [106.92, 169.08] pg/mL, F = 42.64; IL-1β: 59.90 [50.83, 73.78] vs. 151.60 [108.4, 198.36] pg/mL, F = 44.35, all P < 0.01, n = 8). Conversely, IL-10 levels significantly increased (141.84 [118.98, 158.36] vs. 52.96 [42.68, 74.52] pg/mL, F= 126.67, P < 0.01, n = 8). The M2 macrophage count significantly increased (2891.42 ± 211.29 vs. 1583.38 ± 162.22, F= 274.35, P < 0.01 by immunofluorescence test; 0.96 ± 0.18 vs. 0.36 ± 0.05, F= 46.24, P < 0.05 by flowcytometry test). On day 28, cardiac function was better in the ASX group than in the AMI group (LVID_s: 0.50 [0.41, 0.56] vs. 0.64 [0.56, 0.74] cm, Z = -3.60; LVID_d: 0.70 [0.60, 0.76] vs. 0.80 [0.74 0.88] cm, Z = -2.96; ESV: 0.24 [0.18, 0.45] vs. 0.58 [0.44, 0.89] mL, Z = -3.62; EDV: 0.76 [0.44, 1.04] vs. 1.25 [0.82, 1.46] mL, Z = -2.54; EF: 0.60 ± 0.08 vs. 0.50 ± 0.12, F= 160.48; % FS: 0.29 [0.24, 0.31] vs. 0.20 [0.17, 0.21], Z = -4.43, all P < 0.01; n = 16). The MIS and LW/BW ratio were markedly lower in the ASX group than in the AMI group (myocardial infarct size: 32.50 ± 1.37 vs. 50.90 ± 1.73, t = 23.63, P < 0.01, n = 8; LW/BW: 1.81 ± 0.15 vs. 2.17 ± 0.37, t= 3.66, P = 0.01, n = 16). The CVF was significantly lower in the ASX group than in the AMI group: 12.88 ± 2.53 vs. 28.92 ± 3.31, t = 10.89, P < 0.01, n = 8. The expression of caspase 3, TGF-β1, MMP9, and type I/III collagen was lower in the ASX group than in the AMI group (caspase 3: 0.38 ± 0.06 vs. 0.66 ± 0.04, t= 8.28; TGF-β1: 0.37 ± 0.04 vs. 0.62 ± 0.07, t = 6.39; MMP9: 0.20 ± 0.06 vs. 0.40 ± 0.06, t = 4.62; type I collagen: 0.42 ± 0.09 vs. 0.74 ± 0.07, t = 5.73; type III collagen: 0.13 ± 0.02 vs. 0.74 ± 0.07, t = 4.32, all P < 0.01; n = 4).

ASX treatment after AMI may promote M2 macrophages and effectively attenuate cardiac remodeling by inhibiting inflammation and reducing myocardial fibrosis.

Epithelial to mesenchymal transition (EMT) is a key process in determining distant metastasis and intra-hepatic dissemination of hepatocellular carcinoma (HCC). Follistatin (FST) family members are considered to be an attractive therapeutic targets and prognostic indicators in cancers. As a derivative of FST, Follistatin Like 5 (FSTL5) may play a similar role in HCC cells. This study aimed to investigate the expression and function of FSTL5 in HCC and its role in EMT.

FSTL5, E-cadherin and vimentin in HCC, and paracancerous tissues were detected by immunohistochemistry. Correlation of FSTL5 expression with overall survival was assessed. The proliferation and invasion of HCC cell lines SK-Hep1 and MHCC-LM3 were analyzed by cell counting kit-8 and Transwell assays. The expression of FSTL5, E-cadherin, and vimentin in HCC cells was examined by polymerase chain reaction and Western blot analysis. T-test was used to analyze the difference in proliferation and invasion ability between groups. The Spearman rank correlation test was used to detect the correlation between the expression of FSTL5 and E-cadherin or vimentin.

The expression of FSTL5 in HCC was lower than that in paracancerous tissues (9.97% vs. 82.55%, χ² = 340.15, P < 0.001). Patients with high FSTL5 expression had a better prognosis (χ²= 8.22, P= 0.004) and smaller tumor diameter (χ² = 45.52, P < 0.001), less lymph node metastasis (χ² = 5.58, P= 0.02), earlier tumor node metastasis stage (χ² = 11.29, P= 0.001), a reduced number of tumors (χ² = 5.05, P= 0.02), lower alpha-fetoprotein value (χ² = 24.36, P < 0.001), more probability of hepatitis carrying (χ² = 40.9, P < 0.001), and better liver function grade (χ² = 5.21, P = 0.02). Immunohistochemistry showed that FSTL5 expression in HCC tissues was positively correlated with E-cadherin expression (r = 0.38, P < 0.001) and negatively correlated with vimentin expression (r = -0.385, P < 0.001). Furthermore, over-expression of FSTL5 up-regulated the expression of E-cadherin and down-regulated the expression of vimentin in SK-Hep1 (negative control [NC] vs. FSTL5-interfering group [Lv-FSTL5]: E-cadherin [t= 45.03, P < 0.001], vimentin [t= 67, P < 0.001]) and MHCC-LM3 (NC vs. Lv-FSTL5: E-cadherin [t = 50, P < 0.001], vimentin [t = 72.75, P < 0.001]) cells at mRNA level. The same as protein level. In addition, the over-expression of FSTL5 inhibited the proliferation (NC vs. Lv-FSTL5: SK-Hep1, 3 d [t = 7.324, P = 0.018], 4 d [t = 6.23, P = 0.021], 5 d [t= 10.21, P= 0.003]; MHCC-LM3, 3 d [t= 4.32, P= 0.037], 4 d [t= 7.49, P= 0.012], 5 d [t= 9.3661, P = 0.009]) and invasion (NC vs. Lv-FSTL5: SK-Hep1, t= 21.57, P < 0.001; MHCC-LM3, t= 18.04, P < 0.001) of HCC cells.

Down-regulation of FSTL5 may contribute to EMT of HCC, and FSTL5 is a potential target in the treatment of HCC.

We previously found that the intestinal epithelial chemokine (C-C motif) ligand 7 (CCL7) plays an important role in the development of toxin-induced acute liver damage. The detailed effects of intestinal epithelial CCL7 on chronic diseases; however, are still unclear. Here, we aimed to investigate the impact of intestinal epithelial CCL7 overexpression on high-fat diet (HFD)-induced obesity and steatohepatitis in mice.

Intestinal epithelial CCL7 overexpression (CCL7^tgIEC) mice and their wild-type (WT) littermates were fed with normal chow or HFD for 16 weeks to induce obesity and non-alcoholic fatty liver disease. Body weight gain, as well as adipose tissue index were assessed. Liver injury was monitored by histological analysis and real time polymerase chain reaction. Gut microbial composition was analyzed by 16S rRNA gene sequencing.

We found that the CCL7^tgIEC mice on a HFD had markedly decreased weight gain (8.9 vs. 17.0 g, P < 0.05) and a lower adipose tissue index that include mesenteric fat (1.0% vs. 1.76%, P < 0.05), gonadal fat (2.1% vs. 6.1%, P < 0.05), subcutaneous fat (1.0% vs. 2.8%, P < 0.05) compared to WT animals. HFD-induced glucose intolerance and insulin resistance were also significantly improved in CCL7^tgIEC mice compared to WT. Furthermore, HFD-fed CCL7^tgIEC mice displayed less hepatic lipid accumulation and lower expression of inflammatory factors than WT mice. 16S rRNA gene sequencing demonstrated that CCL7 overexpression in intestinal epithelial cells improved HFD-induced gut microbial dysbiosis.

Our study revealed that CCL7 overexpression in the intestinal epithelium protects mice against the progression of diet-induced obesity, hepatic steatosis, and enteric dysbiosis.

Vascular endothelial dysfunction is considered a key pathophysiologic process for the development of acute lung injury. In this study, we aimed at investigating the effects of unfractionated heparin (UFH) on the lipopolysaccharide (LPS)-induced changes of vascular endothelial-cadherin (VE-cadherin) and the potential underlying mechanisms.

Male C57BL/6 J mice were randomized into three groups: vehicle, LPS, and LPS + UFH groups. Intraperitoneal injection of 30 mg/kg LPS was used to induce sepsis. Mice in the LPS + UFH group received subcutaneous injection of 8 U UFH 0.5 h before LPS injection. The lung tissue of the mice was collected for assessing lung injury by measuring the lung wet/dry (W/D) weight ratio and observing histological changes. Human pulmonary microvascular endothelial cells (HPMECs) were cultured and used to analyze the effects of UFH on LPS- or tumor necrosis factor-alpha (TNF-α)-induced vascular hyperpermeability, membrane expression of VE-cadherin, p120-catenin, and phosphorylated myosin light chain (p-MLC), and F-actin remodeling, and on the LPS-induced activation of the phosphatidylinositol-3 kinase (PI3K)/serine/threonine kinase (Akt)/nuclear factor kappa-B (NF-κB) signaling pathway.

In vivo, UFH pretreatment significantly attenuated LPS-induced pulmonary histopathological changes (neutrophil infiltration and erythrocyte effusion, alveolus pulmonis collapse, and thicker septum), decreased the lung W/D, and increased protein concentration (LPS vs. LPS + UFH: 0.57 ± 0.04 vs. 0.32 ± 0.04 mg/mL, P = 0.0092), total cell count (LPS vs. LPS + UFH: 9.57 ± 1.23 vs. 3.65 ± 0.78 × 10⁵/mL, P= 0.0155), polymorphonuclear neutrophil percentage (LPS vs. LPS+ UFH: 88.05% ± 2.88% vs. 22.20% ± 3.92%, P = 0.0002), and TNF-α (460.33 ± 23.48 vs. 189.33 ± 14.19 pg/mL, P = 0.0006) in the bronchoalveolar lavage fluid. In vitro, UFH pre-treatment prevented the LPS-induced decrease in the membrane expression of VE-cadherin (LPS vs. LPS + UFH: 0.368 ± 0.044 vs. 0.716 ± 0.064, P = 0.0114) and p120-catenin (LPS vs. LPS + UFH: 0.208 ± 0.018 vs. 0.924 ± 0.092, P = 0.0016), and the LPS-induced increase in the expression of p-MLC (LPS vs. LPS + UFH: 0.972 ± 0.092 vs. 0.293 ± 0.025, P = 0.0021). Furthermore, UFH attenuated LPS- and TNF-α-induced hyperpermeability of HPMECs (LPS vs. LPS + UFH: 8.90 ± 0.66 vs. 15.84 ± 1.09 Ω·cm², P = 0.0056; TNF-α vs. TNF-α + UFH: 11.28 ± 0.64 vs. 18.15 ± 0.98 Ω·cm², P = 0.0042) and F-actin remodeling (LPS vs. LPS + UFH: 56.25 ± 1.51 vs. 39.70 ± 1.98, P = 0.0027; TNF-α vs. TNF-α + UFH: 55.42 ± 1.42 vs. 36.51 ± 1.20, P = 0.0005) in vitro. Additionally, UFH decreased the phosphorylation of Akt (LPS vs. LPS + UFH: 0.977 ± 0.081 vs. 0.466 ± 0.035, P = 0.0045) and I kappa B Kinase (IKK) (LPS vs. LPS + UFH: 1.023 ± 0.070 vs. 0.578 ± 0.044, P = 0.0060), and the nuclear translocation of NF-κB (LPS vs. LPS + UFH: 1.003 ± 0.077 vs. 0.503 ± 0.065, P = 0.0078) in HPMECs, which was similar to the effect of the PI3K inhibitor, wortmannin.

The protective effect of UFH against LPS-induced pulmonary endothelial barrier dysfunction involves VE-cadherin stabilization and PI3K/Akt/NF-κB signaling.

Acquired aplastic anemia (AAA) is an auto-immune disease (AID) resulting from aberrant T-cell-mediated and antigen-driven immune responses to hematopoietic stem and progenitor cells (HSPCs) in genetically susceptible individuals, leading to the significantly enhanced suppression and apoptosis of HSPCs. Active systemic and local inflammation is responsible for the overall pathophysiology. The dysregulated auto-immunity is characterized by the abnormally increased number and function of proinflammatory cells and the abnormally decreased immunoregulatory cells. Significantly up-regulated expression of human leukocyte antigen (HLA) and Fas molecules on CD34⁺HSPCs induces enhanced antigen-presenting activities and accelerated apoptosis.^[1]The high efficiency of rapamycin in ameliorating bone marrow suppression (BMS) clearly indicates the abnormally activated mTOR (mammalian target of rapamycin) pathway and its contribution.

To the Editor:Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide. It is associated with visceral obesity and is considered to be the hepatic component of metabolic syndrome, as insulin resistance plays a major role in this process. Despite its known association, the pathogenesis remains uncertain. Recent works from both our groups and other groups have demonstrated that zinc-α2-glycoprotein (ZAG) or alpha-2-glycoprotein 1, zinc-binding, a novel adipokine, is closely linked to obesity and obesity-related metabolic disease. Because of this, it has been proposed as a candidate factor in the regulation of lipid metabolism and insulin sensitivity.^[1]Despite its link, the clinical relevance of ZAG in NAFLD remains largely unknown. We conducted a case-control study to measured serum ZAG levels as well as its association with various metabolic parameters in NAFLD patients.

To the Editor:Colorectal cancer (CRC) remains the second most frequent cause of cancer-related deaths worldwide. Since the concept of total mesorectal excision (TME) introduced by Healdet al^[1]has become a gold standard of rectal cancer surgery, there has been a significant improvement in oncological outcomes including disease-free and overall survival. Although TME is the standard procedure for rectal cancer patients undergoing either low anterior resection (LAR) or abdominoperineal resection, the optimum strategy to restore bowel continuity between colon and rectum (in case of LAR) is still debated. There is no clear consensus of the best approach both in terms of anastomotic technique^[2](end-to-end or side-to-end) and in terms of vessel ligation (high tie or low tie ligation).

To the Editor:Sepsis is defined as the life-threatening organ dysfunction caused by dysregulated host response to infection, eventually leading to multiple organ dysfunction syndrome. The gut, which is considered to play a significant role during sepsis, is defined as the motor of sepsis. During sepsis, increasing epithelial apoptosis damages gut integrity and weakens cell adherence, eventually leading to intestinal barrier failure and aggravating the degree of sepsis.^[1]The nuclear factor (NF)-κB pathway plays an important role in intestinal mucosal barrier failure during sepsis. However, whether NF-κB activation plays a harmful or beneficial role in the intestine in response to inflammation remains a matter of debate.^[2]Here, we established a model of lipopolysaccharide (LPS)-induced intestinal sepsisin vitroand measured the apoptosis in sepsis intestinal tissues to explore the function of NF-κB pathway in sepsis.

To the Editor:In the very interesting report published in theChinese Medical Journal,^[1]a 42-year-old Chinese male patient who had not any previous history of cardiovascular disease, but only bee stings without any sequelae, was stung again while was cycling outdoors. Immediately, he developed sweats, dizziness, headaches, bosom frowsty, shortness of breath, urinary incontinence, hypotension, hypoxygenation, tachycardia, confusion, ST-segment depression in II-III leads, and ST-segment elevation in aVR, eosinophilia, increased cardiac enzymes, but with normal coronary arteries on angiography, compatible with Kounis type I variant progressing to acute myocardial infarction.^[2,3]In this patient; however, specific immunoglobulin tests were positive for tree combination (willow/poplar/elm). The patient had complete recovery with antiallergic medications and fluid replacement.