"One-gate" design should be used for the study of the diagnostic efficacy of biological markers.

1. The first draft of this article proposes "Please authors answer a question: was there only 43 patients with diagnosed gastric cancer from April 2018 to July 2018?" Is it to know whether the researchers chose 43 patients because of fewer cases or intentionally fewer? The reviewer's "response reviewers" admitted "it's better to add a benign disease group" and "chose 43 patients with diagnosed gastric cancer randomly for our validation," which confirmed my opinion "have design flaws both in enroll subjects and the number of cases". So after getting the reply from the original author, I deleted the above sentence. This time amend "43 patients" on line 43 to "43 patients which random selection rather than all patients in the study time window". 2. Some revision has been highlighted in red. 3. We re-edit the article sentence by sentence in English to make it more in line with the English expression.

Moxibustion Reduces Inflammatory Response in the Hippocampus of a Chronic Exercise-Induced Fatigue Rat.

Accumulating data indicates that brain inflammation plays an important role in the pathophysiology of chronic exercise-induced fatigue. Moxibustion in traditional Chinese medicine has been found to alleviate exercise-induced fatigue. However, it remains unclear whether the effect of moxibustion is related to its anti-inflammatory properties. In this study, rats were exposed to 3-week exhaustive swimming to induce chronic exercise-induced fatigue. The body weight, exhaustive swimming time, tail suspension test and open-field test were observed. Real-time polymerase chain reaction (RT-PCR) was used to determine the mRNA expression of proinflammatory cytokines (interleukin-1ß [IL-1ß], interleukin-6 [IL-6], and tumor necrosis factor-α[TNF-α]), and enzyme-linked immunosorbent assay (ELISA) was used to detect IL-1ß, IL-6, and TNF-α concentrations. Chronic exhaustive exercise significantly reduced the body weight and exhaustive swimming time, and increased tail suspension immobility time, which were reversed by moxibustion treatment. Compared with control rats, the mRNA and protein expression of IL-1ß, IL-6, and TNF-α in the hippocampus was significantly increased in exhaustive swimming trained rats. Moxibustion significantly decreased the level of IL-6 in the hippocampus, but not affected IL-1ß and TNF-α level significantly. Our results suggested that a potential inflammatory damage in the brain may be involved during chronic exhaustive exercise-induced fatigue. Moxibustion could attenuate the inflammatory impairment in exercise-induced fatigue, which might be mediated by inhibition of the proinflammatory cytokine IL-6 levels in the brain region.

[Effect of moxibustion on inflammatory cytokine levels in hippocampus and hypothalamus in rats with fatigue].

OBJECTIVE: To investigate the effect of moxibustion of "Zusanli "(ST36) on levels of inflammatory cytokines in the hippocampus and hypothalamus regions of rats with fatigue, so as to reveal its anti-fatigue mechanism. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into normal control, fatigue model, and moxibustion groups, with 8 rats in each group. The fatigue model was established by chronic weight-loaded exhaustive swimming. During modeling, the rats in the moxibustion group were given moxibustion (3 moxa cones, about 8 min in duration) at bilateral ST36, once every other day for 11 times in total. The duration of exhausted swimming was observed, and ELISA was used to measure the contents of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interferon gamma (IFN-γ) in the hippocampus and hypothalamus tissues. RESULTS: The duration of exhausted swimming was obviously prolonged on day 14 and 21 after moxibustion intervention relevant to the model group (P<0.05, P<0.01). The contents of IL-1ß， IL-6 and TNF-α in the hippocampus were significantly increased in the model group than in the normal group (P<0.01), and that of hippocampal IL-6 was considerably decreased in the moxibustion group than in the model group (P<0.05). The content of hippocampal IFN-γ was significantly lower in the model group than in the normal group (P<0.05) and was increased mildly after moxibustion (P>0.05). No significantly changes were found in the levels of hypothalamic IL-1ß， IL-6, TNF-α and IFN-γ after modeling and moxibustion (P>0.05), and in the levels of hippocampal IL-1ß， TNF-α and IFN-γ after moxibustion (P>0.05)．. CONCLUSION: Moxibustion can significantly down-regulate hippocampal IL-6 content in fatigue rats, which maybe contribute to its effect in alleviating fatigue. Further studies need being conducted to identify this conclusion.

Effects of Mongolian Warm Acupuncture on iNOS/NO and Inflammatory Cytokines in the Hippocampus of Chronic Fatigue Rats.

The inducible nitric oxide synthase/nitric oxide (iNOS/NO) signaling pathway and inflammatory cytokines play important roles in the pathogenesis of exercise-induced fatigue. Studies have found that Mongolian warm acupuncture (WA) could alleviate exercise-induced fatigue. However, the exact mechanisms underlying its effects remain unclear. In the present study, we investigated the effects of Mongolian WA on iNOS/NO signaling pathway and proinflammatory cytokines in a chronic exhaustive swimming-induced fatigue rat model. Animals were randomly divided into Control group, Ctrl + WA group, Model group, and Model + WA group. The body weight, exhaustive swimming time test, and Morris water maze test were performed before and after the chronic exhaustive swimming. The serum levels of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and iNOS were detected by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of IL-1ß, IL-6, TNF-α, IFN-γ, and iNOS in the hippocampus were measured by real-time polymerase chain reaction (RT-PCR). Moreover, the protein expression of iNOS in the hippocampus was measured by western blot, and the NO productions in the serum and hippocampus were detected by Griess reaction system. Chronic exhaustive exercise significantly reduced the body weight and exhaustive swimming time, and induced impairment in learning and memory, and which were reversed by WA treatment. Chronic exhaustive exercise also increased the expressions of iNOS and proinflammatory cytokines, while WA treatment significantly decreased the level of iNOS and proinflammatory cytokines. However, chronic exhaustive exercise did not affect the NO production. These findings demonstrated that WA could alleviate the chronic exhaustive swimming-induced fatigue and improve the learning and memory ability, and the actions might be related to the reduction of inflammatory response and iNOS expression.

A Novel Method for Estimating Low-Density Lipoprotein (LDL) Levels: Total Cholesterol and Non-High-Density Lipoprotein (HDL) Can Be Used to Predict Abnormal LDL Level in an Apparently Healthy Population.

BACKGROUND We aimed to predict the abnormal LDL level by using TG, TC, HDL, and non-HDL in this study. MATERIAL AND METHODS Triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) data were obtained from the Laboratory Information System (LIS) for 4 years (Oct 1, 2013 to Sept 30, 2017) from among 34 270 healthy Chinese patients at Shuyang People's Hospital. TG, TC, HDL, and LDL (direct clearance method) were measured using a TBA2000FR biochemical analyzer. The non-HDL was calculated as TC minus HDL. Correlations between TG, TC, non-HDL, and LDL were analyzed using Spearman's rank correlation. Receiver operating characteristics (ROC) curve analysis was used to evaluate the predictive utility of TG, TC, and non-HDL for the abnormal LDL level (<130 mg/dL). RESULTS Both TC (r=0.870) and non-HDL (r=0.893) were significantly positively correlated with LDL. The area under curve of TC and non-HDL can be used to predict abnormal LDL levels. Optimal thresholds were 182.5 mg/Dl (4.72 mmol/L) for TC and 135.3 mg/Dl (3.50 mmol/L) for non-HDL. Based on these optimal thresholds, less than 0.5% and 0.4% of tests with elevated LDL were missed using TC and non-HDL, respectively, but the value of these missed LDL levels was not very high (<147.3 mg/dL). CONCLUSIONS If the value of non-HDL is less than 135.3 mg/Dl (3.50 mmol/L) and/or TC is less than 182.5 mg/Dl (4.72 mmol/L) for the apparently healthy populations, the LDL level will be less than 130 mg/Dl (3.36 mmol/L). TC and non-HDL can be used to predict the abnormal LDL level in apparently healthy populations.

Reference intervals of carbohydrate antigen 19-9 in the apparently healthy adult population.

BACKGROUND: To establish reference intervals of carbohydrate antigen 19-9(CA 19-9) according to the CLSI CA28-A3 guideline and to evaluate age- and gender-related variations. METHODS: Serum CA 19-9 values of 10 149 healthy subjects (from 20 years old to 60 years old) were measured from location health checkups. The relationship between CA 19-9 and age was analyzed using Spearman's approach. The reference intervals of CA19-9 were established using Q2.5 and Q97.5 , and the 90% confidence intervals of upper limits were calculated. RESULTS: The reference intervals of CA 19-9 were 1.98-25.12 U/mL for males (1.97-25.06 U/mL for 20-50 years old and 2.31-26.13 U/mL for 50-60 years old) and 2.36-29.29 U/mL for adult (20-60 years old) females. The upper limit of reference intervals for all individuals was 26.45 U/mL; the level of CA 19-9 is higher in females than males. Carbohydrate antigen (CA) 19-9 is significantly associated with aging in adult males(r = .0930, P < .0001), but not in females (P = .4734). CONCLUSIONS: Establishing reference intervals for CA19-9 and giving age-related reference intervals of CA19-9 using a big data of healthy adult, we first discovered that CA19-9 tends to increase with age in adult males but not in females.

Establishing Reference Intervals of Aspartate Aminotransferase-to-Platelet Ratio Index for Apparently Healthy Elderly.

BACKGROUND: Aspartate aminotransferase (AST) to platelet ratio index (APRI) serves as a parameter in evaluating liver fibrosis in current clinical practice. However, reference standard (reference intervals, RIs) or baseline levels of APRI have not been previously reported. The purpose of this paper is to establish the reference intervals of APRI in apparently healthy elderly people from the region of Shuyang, China. METHODS: Blood specimens were collected from local elderly residents (selected 51,263 elderly Han Shuyang Chinese from 65 to 97 years old, 32.97% males and 67.03% females) by standard procedures. Complete blood counts were determined by Sysmex XE-2100 analyzer and the AST values were measured by a TBA2000FR automatic biochemical analyzer (Toshiba Co., Ltd., Japan). The 95% reference intervals were calculated by using the non-parametric method according to the document: Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline-Third Edition (C28-A3) of CLSI. RESULTS: RIs established for healthy elderly include: 0.1398-0.6266 for males and 0.1282-0.5798 for females (0.1284-0.5086 for 65-74 years old; 0.1209-0.5704 for > or = 75 years old). Ris of APRI for elderly males were higher than those of females, and values of APRI increased with increasing age for females. CONCLUSIONS: We established scientific and reasonable RIs of APRI for the healthy elderly in our region.

Comparative proteomics analysis of cerebrospinal fluid of patients with Guillain-Barré syndrome.

To better understand the pathophysiologic mechanisms underlying Guillain-Barré syndrome (GBS), Comparative proteomic analysis of cerebrospinal fluid (CSF) between patients with GBS (the experiment group) and control subjects suffering from other neurological disorders (the control group) was carried out using two-dimensional gel electrophoresis (2-DE) technique, in combination with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) and database searching to determine abnormal CSF proteins in GBS patients. Image analysis of 2-DE gels silver stained revealed that 10 protein spots showed significant differential expression between the two groups of CSF samples. The expression of cystatin C, transthyretin, apolipoprotein E and heat shock protein 70 were decreased. However, haptoglobin, alpha-1-antitrypsin, apolipoprotein A-IV and neurofilaments were elevated. The subsequent ELISA measured the concentration of cystatin C and confirmed the result of the proteomic analysis. These identified proteins may be involved in the pathophysiological process of GBS and call for further studying the role of these proteins in the pathogenesis of the disease.