[Effect of Erxian Decoction-containing serum on H_2O_2-induced proliferation and osteogenic differentiation of MC3T3-E1 cells via BK channels].

This study aimed to investigate the effects of Erxian Decoction(EXD)-containing serum on the proliferation and osteogenic differentiation of MC3T3-E1 cells under oxidative stress through BK channels. The oxidative stress model was induced in MC3T3-E1 cells by H_2O_2, and 3 mmol·L~(-1) tetraethylammonium(TEA) chloride was used to block the BK channels in MC3T3-E1 cells. MC3T3-E1 cells were divided into a control group, a model group, an EXD group, a TEA group, and a TEA+EXD group. After MC3T3-E1 cells were treated with corresponding drugs for 2 days, 700 µmol·L~(-1) H_2O_2 was added for treatment for another 2 hours. CCK-8 assay was used to detect cell proliferation activity. The alkaline phosphatase(ALP) assay kit was used to detect the ALP activity of cells. Western blot and real-time fluorescence-based quantitative PCR(RT-qPCR) were used to detect protein and mRNA expression, respectively. Alizarin red staining was used to detect the mineralization area of osteoblasts. The results showed that compared with the control group, the model group showed significantly blunted cell proliferation activity and ALP activity, reduced expression of BK channel α subunit(BKα), collagen Ⅰ(COL1), bone morphogenetic protein 2(BMP2), osteoprotegerin(OPG), and phosphorylated Akt, decreased mRNA expression levels of Runt-related transcription factor 2(RUNX2), BMP2, and OPG, and declining area of calcium nodules. EXD-containing serum could significantly potentiate the cell proliferation activity and ALP activity, up-regulate the protein expression of BKα, COL1, BMP2, OPG, and phosphorylated Akt, and forkhead box protein O1(FoxO1), promote the mRNA expression of RUNX2, BMP2, and OPG, and enlarge the area of calcium nodules. However, BK channel blockage by TEA reversed the effects of EXD-containing serum in promoting the protein expression of BKα, COL1, BMP2, OPG, and phosphorylated Akt and FoxO1, increasing the mRNA expression of RUNX2, BMP2, and OPG, and enlarging the area of calcium nodules. EXD-containing serum could improve the proliferation activity, osteogenic differentiation, and mineralization ability of MC3T3-E1 cells under oxidative stress, which might be related to the regulation of BK channels and downstream Akt/FoxO1 signaling pathway.

Recent Advancements in Prevention and Treatment of Osteoporosis with Traditional Chinese Medicine: A Long Way from Lab Bench to Bedside.

Osteoporosis is becoming more prevalent in the ageing society, however, its treatment is still a problem for both society and individuals. Traditional Chinese Medicine (TCM) has a long history in treating osteoporosis and is receiving increasing attention. Multiple formulas of TCM showed satisfactory effects in treating osteoporosis in both animal models and clinical patients. However, because TCM usually consists of multiple plant and/or animal products, it is difficult to clarify the mechanism of TCM according to the requirements of Western medicine regarding purity, efficacy, dosage, and safety. With increasing researchers have started to investigate the TCM using modern scientific tools such as bioinformatics and network pharmaceutics in osteoporosis and the addition of TCM in the latest version of International Statistical Classification of Diseases and Related Health Problems (ICD-11 version, 2019) by WHO, TCM is showing large potential in treating osteoporosis although there is still a long way. The review aimed to summarize recent advancements of TCM treating osteoporosis.

[Effects of long-term moderate-small intensity aerobic exercise on differential expression of proteome in left ventricular muscle of rats].

Objective: To investigate the effects of long-term moderate-small intensity aerobic exercise on the differential expression of proteome in left ventricular muscle of rats, and to screen the target proteins sensitive to moderate-small intensity aerobic exercise stimulation. This study will enrich the basic theory of exercise and fitness and provide new ideas and experimental basis for the rehabilitation treatment of chronic cardiovascular disease. Methods: Twenty male SD rats were randomly divided into exercise group and control group (n=10). The treadmill training model of long-term moderate-small intensity aerobic exercise was established, and the whole protein samples of left ventricular muscle were extracted and separated by two-dimensional gel electrophoresis (2-DE). The two-dimensional gel electrophoresis map was analyzed by Bio PD quest image analysis software. The protein spots with differential expression more than 5 times or down-regulated over 80% after exercise were identified by tandem time-of-flight mass spectrometry (ULGRAFL-FLEX-TOF/TOF). Results: Compared with the group C, the heart weight index of the group E was increased by 32.0%, and the difference was significant (P＜0.05). Compared with the group C, there were 71 protein spots expression were up-regulated≥2 times or down-regulated≥50% in the group E. 4 protein spots expression were up-regulated≥5 times or down-regulated≥80% were identified by mass spectrometry, 3 proteins and 1 unknown protein were identified. Conclusion: After long-term moderate-small intensity aerobic exercise, the rats heart had a good adaptive change, and the proteome of left ventricular muscle changed significantly. Long-term moderate-small intensity aerobic exercise can effective enhance the ability of myocardial antioxidation.

[Effect of Gancao Fuzi Decoction on osteoarthritis and proteomics of articular cartilage in rats].

The rat osteoarthritis model was replicated by injection of sodium iodoacetate into the knee joint cavity, and the effects of Gancao Fuzi Decoction on rat osteoarthritis and the proteome of articular cartilage were investigated. Sixty SD rats weighing 230-250 g were randomly divided into normal group, model group, glucosamine sulfate group, and Gancao Fuzi Decoction high, medium and low dose groups. Osteoarthritis model was induced by intra-articular injection of sodium iodoacetate(3 mg on each leg) in all groups except the normal group. After modeling, each administration group was given intragastric administration for 1 month. During the administration period, joint pain test and joint width measurement were performed every week to observe the autonomous behavior of rats. Enzyme linked immunosorbent assay(ELISA) method was used to detect the contents of tumor necrosis factor-α(TNF-α), interleukin-1ß(IL-1ß), matrix metalloproteinase-3(MMP-3), and matrix metalloproteinase tissue inhibitor(TIMP-1) in rat joint lavage fluid. Hematoxylin-eosin(HE) staining was used to observe bone and joint morphology. Nano-LC-LTQ-Orbitrap system was used to detect arti-cular cartilage proteins. The results showed that, compared with the model group, Gancao Fuzi Decoction could significantly improve joint pain and joint swelling in osteoarthritis rats, significantly reduce the contents of TNF-α, IL-1ß and MMP-3 in the joint cavity la-vage fluid, increase the content of TIMP-1, and relieve inflammatory diseases such as enlarged joint space, rough cartilage edge, different thickness of cartilage layer, and disordered arrangement of chondrocytes. After comparing the proteins between the groups, 273 differential proteins were screened out. KEGG analysis found that the above differential proteins involved 43 signaling pathways such as systemic lupus erythematosus, among which 11 signaling pathways were related to osteoarthritis. The above results indicated that Gancao Fuzi Decoction had a preventive effect on osteoarthritis, and its mechanism of action may be accomplished by regulating the protein expression of osteoarthritis-related signal pathways.

[Effect of acupuncture and moxibustion on post-stroke constipation: a network Meta-analysis].

OBJECTIVE: To systematically review the therapeutic effect of acupuncture-moxibustion therapies on post-stroke constipation based on the network Meta-analysis. METHODS: The randomized controlled trials of acupuncture and moxibustion for post-stroke constipation were retrieved from the databases, starting from the time of establishment through to June, 2019, i.e. the PubMed, EMbase, Cochrane Library, Web of Science, SinoMed, CNKI, Wanfang and VIP. The literature was selected according to inclusion and exclusion criteria, the quality of literature was evaluated by bias risk assessment tool of Cochrane Review Manual 5.3 and the data was statistically analyzed by softwares of Stata 13.1 and R Language. RESULTS: A total of 28 trails were included, involving 9 intervention methods. The therapeutic effect was arranged from high to the low according to the surface under cumulative ranking area (SUCRA), i.e. acupuncture combined with medication (0.86), warm needling (0.83), electroacupuncture combined with medication (0.68), electroacupuncture (0.68), moxibustion (0.50), auricular point sticking (0.44), acupuncture (0.31), Chinese herbal medicine (0.12) and western medication (0.04). CONCLUSION: Acupuncture- moxibustion therapies have superiority on treating post-stroke constipation, acupuncture combined with medication has the most optimum therapeutic effect and the therapeutic effect of acupuncture-moxibustion combined with medication is superior to the single treatment of medication.

[Study on 4 weeks medium intensity aeraobic exercise induce the difference expression of the proteomics in rat atrial muscle].

OBJECTIVE: To investigate the effect of moderate-intensity aerobic exercise on the differential expression of rat atrial muscle Proteomics and genes, which provide research basis for the rehabilitation of chronic cardiovascular diseases and exercise -induced cardiac remodeling research. METHODS: Twenty male SD rats were randomly divided into control group and experimental group (n=10) according to body weight. Rats in the experimental group were trained (6 days per week),which lasted for 4 weeks of moderate-intensity aerobic exercise at a rate of 24 m·min-1 for 40 min (load intensity equivalent to 60%~70% VO2max). The proteins were separated by two dimensional gel electrophoresis, and the tandem time-of- flight mass spectrometer technique was used to identify 13 candidate target protein spots. The expression levels of these 13 protein spots were up-regulated more than 5 times or down -regulated to below 1/5. The mRNA of six target proteins were detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: By software analysis, the experimental group compared with the control group, there were 8 protein points which their expression reduced more than 4/5 and 5 protein points up-regulated more than 5 times, 13 proteins were identified by mass spectrometry protein spots, the final identification results acquired 8 proteins and a unknown protein of molecular mass 54 KDa, such as:pyruvate dehydrogenase E1α1, mitochondrial aconitate hydratase, protein disulfide isomerase A3, methylmalonic acid semialdehyde dehydrogenase, mitochondrial dihydrolipoic acid dehydrogenase, isovaleryl coenzyme A dehydrogenase, glutathione synthetase, mitogen-activated protein kinase 3 and so on. Compared with the control group, the mRNA expression of methylmalonic acid semialdehyde dehydrogenase in the atrial muscle of rats was decreased after 4 weeks of moderate aerobic exercise (P<0.05), the mRNA expression levels of mitochondria Ⅱ lipoic acid dehydrogenase, protein disulfide isomerase A3, mitochondrial aconitate hydratase and glutathione synthetase were decreased (P>0.05); The mRNA expression level of isopentenyl-CoA dehydrogenase was increased (P>0.05). The results indicated that the mRNA expression level was not completely consistent with the changes in mass spectrometry identification results. CONCLUSIONS: The 4 weeks moderate-intensity aerobic exercise induced ignificant changes of rats atrial muscle protemics. The majority of the 13 identified target proteins in this experiment are energy metabolism enzymes. The majority of the expression of the target protein and the mRNA expression in the atrial muscle is inconsistent and different. Exercise may affect the regulation of gene transcription or downstream translation and modification of these target proteins, resulting in the change of differential expression.

Analysis of islet beta cell functions and their correlations with liver dysfunction in patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD).

Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) primarily manifests in neonates or infants with hepatomegaly, liver dysfunction, and hypoglycemia. This study investigated the functions of islet beta cells and their correlations with liver dysfunction in NICCD patients.We retrospectively analyzed clinical data on liver function and islet beta cell functions for 36 patients diagnosed with NICCD and 50 subjects as the control group. The NICCD group had significantly higher total bilirubin (TBIL), direct bilirubin (DBIL), alanine aminotransferase (ALT), aspartate amino transferase (AST), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP) and alpha-fetoprotein (AFP) levels and albumin/globulin ratio (A/G) (P < .05), and lower ALB and GLB levels than the control group (P < .05). The differences in fasting blood glucose (FBG), fasting insulin, C-peptide (C-P), the homeostasis model of assessment for the insulin resistance index (HOMA-IR), fasting beta cell function (FBCI), and the HOMA beta cell function index (HBCI) between the NICCD and control groups were not significant (P > .05). A linear correlation was found between FBG and fasting insulin (P < .001) and between FBG and C-P in the NICCD patients (P = .001). Fasting insulin (P = .023), HOMA-IR (P = .023), FBCI (P = .049), and HBCI (P = .048) were positively correlated with increases in the ALT level. There was no difference in islet beta cell functions between the NICCD and control groups. The liver dysfunction may be correlated with islet beta cell functions in NICCD patients.

A Systematic Review and Meta-Analysis of Influenza A Virus Infection During Pregnancy Associated with an Increased Risk for Stillbirth and Low Birth Weight.

BACKGROUND/AIMS: Impaired pregnancy outcomes, such as low birth weight are associated with increased disease risk in later life, however little is known about the impact of common infectious diseases during pregnancy on birth weight. The study had two aims: a) to investigate risk factors of influenza virus infection during pregnancy, and b) to analyze the impact of influenza virus infection on pregnancy outcome, especially birth weight. METHODS: Prospective and retrospective observational studies found in PubMed, MEDLINE, Embase, Google Scholar, and WangFang database were included in this meta analysis. Data of included studies was extracted and analyzed by the RevMan software. RESULTS: Pregnant women with anemia (P=0.004, RR=1.46, 95% CI: 1.13-1.88), obesity (P<0.00001, RR=1.35, 95% CI: 1.25-1.46) and asthma (P<0.00001, RR=1.99, 95% CI: 1.67-2.37) had higher rates of influenza virus infection. Regarding birth outcomes, influenza A virus infection did not affect the likelihood for cesarean section. Mothers with influenza had a higher rate of stillbirth (P=0.04, RR=2.36, 95% CI: 1.05-5.31), and their offspring had low 5-minute APGR Scores (P=0.009, RR=1.39, 95% CI: 1.08-1.79). Furthermore, the rate for birth weight < 2500g (P=0.04, RR=1.71, 95% CI: 1.03-2.84) was increased. CONCLUSION: Results of this study showed that anemia, asthma and obesity during pregnancy are risk factors influenza A virus infection during pregnancy. Moreover, gestational influenza A infection impairs pregnancy outcomes and increases the risk for low birth weight, a known risk factor for later life disease susceptibility.

Epigenetic modulation of insulin-like growth factor-II overexpression by hepatitis B virus X protein in hepatocellular carcinoma.

Hepatitis B virus X protein (HBx) is involved in the pathogenesis of hepatocellular carcinoma (HCC). Overexpression of the transcripts from the P3 and P4 promoters of the insulin-like growth factor-II (IGF-II) gene is observed in HCC. The present study investigated the involvement of HBx in IGF-II overexpression and its epigenetic regulation. Firstly, the effects of HBx on P3 and P4 mRNA expression, the methylation status of the P3 and P4 promoters, and MBD2 expression were analyzed in human HCC cells and HCC samples. Next, interaction between HBx and MBD2 or CBP/p300 was assessed by co-immunoprecipitation, and HBx-mediated binding of MBD2 and CBP/p300 to the P3 and P4 promoters and the acetylation of the corresponding histones H3 and H4 were evaluated by quantitative chromatin immunoprecipitation. Finally, using siRNA knockdown, we investigated the roles of MBD2 and CBP/p300 in IGF-II overexpression and its epigenetic regulation. Our results showed that HBx promotes IGF-II expression via inducing the hypomethylation of the P3 and P4 promoters, and that HBx increases MBD2 expression, directly interacts with MBD2 and CBP/p300, and elevates their recruitment to the hypomethylated P3 and P4 promoters with increased acetylation levels of the corresponding histones H3 and H4. Further results showed that endogenous MBD2 and CBP/p300 are necessary for HBx-induced IGF-II overexpression and that CBP/p300 presence and CBP/p300-mediated acetylation of histones H3 and H4 are partially required for MBD2 binding and its demethylase activity. These data suggest that HBx induces MBD2-HBx-CBP/p300 complex formation via interaction with MBD2 and CBP/p300, which contributes to the hypomethylation and transcriptional activation of the IGF-II-P3 and P4 promoters and that CBP/p300-mediated acetylation of histones H3 and H4 may be a rate-limiting step for the hypomethylation and activation of these two promoters. This study provides an alternative mechanism for understanding the pathogenesis of HBx-mediated HCC.

Telbivudine during the second and third trimester of pregnancy interrupts HBV intrauterine transmission: a systematic review and meta-analysis.

UNLABELLED: BECKGROUND: Evaluate the efficacy and safety of telbivudine during the 2nd and 3rd trimester of pregnancy in intrauterine transmission of hepatitis B virus (HBV). Based on the principle of Cochrane systematic reviews, a database was constructed from Medline, EMBASE, Cochrane Library, the US National Science Digital Library (NSDL), the China Biological Medicine Database (CBM-disc), and contact with Chinese experts in the field from November 2006 to February 2013. METHODS: The Critical Appraisal Skills Programme (CASP) of Oxford, Cochrane Collaboration's tool, and Review Manager Version 5.0 (Rev-Man 5.0) for assessing the quality of clinical trials, risk of bias, and statistical analysis was used. We analyzed the effects and safety of telbivudine treatment on intrauterine mother-to-child transmission (MTCT) of HBV from HBsAg and HBV-DNA positive mothers. All newborns received an immune prophylaxis schedule consisting of simultaneous hepatitis B virus vaccine and hepatitis B immunoglobulin (HBIG) postpartum. Of 32 studies, 7 studies fulfilled the inclusion criteria in the study. RESULTS: Either the Mantel-Haenszel or Inverse Variance fixed-effects model or Mantel-Haenszel or Inverse Variance random-effects model was applied for all analyses indicated by odds ratio (OR) and 95% confidence interval (CI). The meta-analysis based on new onset of HBsAg seropositivity of infants at 6-12 months postpartum revealed that the control group had an intrauterine transmission rate of 8.25-42.31%. This rate was reduced to 0-14.29% in the telbivudine treatment group (OR 0.09, 95% CI 0.04-0.22, including seven trials, p < 0.001). The rates of intrauterine transmission based on new onset of HBV DNA seropositivity of infants at 6-12 months postpartum were 8.25-19.23% in the control group and 0 - 3.57% in the treatment group (OR 0.07, 95% CI 0.02-0.22, p < 0.001, including only five trials, since two trials had no data on HBV DNA in infants). With the exception of CK elevations, adverse effect frequencies were similar in both groups. CONCLUSIONS: Telbivudine is an effective and safe drug for preventing intrauterine transmission of HBV.

Hepatic impairment induced by scrub typhus is associated with new onset of renal dysfunction.

BACKGROUND: Scrub typhus is a potentially fatal infectious disease caused by Orientia tsutsugamushi. There is little attention given to hepatic impairment in the adults with scrub typhus. This study investigated the incidence and the prognostic implications of hepatic impairment in patients with scrub typhus. METHODS: We retrospectively reviewed a total of 143 adult patients with scrub typhus who were admitted between January 1999 and December 2010 in Guangdong province, China. The patients were divided into three groups, e.g., normal, mild, and moderate to severe groups based on the elevated serum ALT and/or total bilirubin levels. Furthermore, clinical characteristics and prognosis of the patient groups were compared. RESULTS: 109 patients (76.2%) had abnormal liver function. Among the patients with hepatic impairment 45 cases (31.4%), 54 cases (37.8%), and 10 cases (7.0%) had mild, moderate, and severe hepatic damage, respectively. The moderate to severe hepatic impairment group had higher levels of serum creatinine compared with that of normal hepatic function. The incidence of new onset of renal dysfunction - defined as peak serum creatinine > or = 176 micromol/L during hospital stay with no evidence of renal disease prior hospitalization - was 0% in the mild hepatic impairment group, 8.9% in the moderate hepatic impairment group, and 21.9% in the severe hepatic impairment group, (p = 0.005 for trend). Additionally, the patients with hepatic impairment (n = 109) had higher incidences of episodes of thrombocytopenia (45.9% vs. 8.82%, p < 0.001), hypoalbuminemia (50.5% vs. 11.8%, p < 0.001), new onset of renal dysfunction (16.5% vs. 0.0%, p = 0.011), and electrocardiogram abnormality (28.4% vs. 8.82%, p = 0.019) than the patients without hepatic impairment. CONCLUSIONS: The degree of hepatic impairment induced by scrub typhus is associated with new onset of renal dysfunction.

Low birth weight is associated with lower respiratory tract infections in children with hand, foot, and mouth disease.

BACKGROUND: Low birth weight (LBW) might be a risk factor for acquiring lower respiratory tract infections (LRTIs) associated with disease related complications in early childhood. HFMD, a frequent viral infection in southern China, is a leading cause of lower respiratory tract infections in children. We analyzed whether LBW is a risk factor for children with HFMD to develop lower respiratory tract infections. METHODS: A total of 298 children with HFMD, admitted to a hospital in Qingyuan city, Guangdong province, were recruited. Demographic data and clinical parameters such as serum glucose level and inflammatory markers including peripheral white blood cell count, serum C-reactive protein, and erythrocyte sedimentation rate were routinely collected on admission. Birth weight data were derived from birth records. RESULTS: Mean birth weight (BW) was 167 g lower in patients with HFMD and LRTIs as compared to patients with solely HFMD (p = 0.022) and the frequency of birth weight below the tenth percentile was significantly higher in patients with HFMD and LRTIs (p = 0.002). CONCLUSIONS: The results of the study show that low birth weight is associated with a higher incidence of lower respiratory tract infections in young children with HFMD.

Differentiation of human umbilical cord mesenchymal stem cells into hepatocyte-like cells by hTERT gene transfection in vitro.

The availability of a large quantity of MSCs (mesenchymal stem cells) would greatly advance liver-directed cell therapies. However, MSCs have a limited lifespan in vitro. Therefore we tested whether hUCMSCs (human umbilical cord MSCs) could be immortalized by transduction with a lentiviral vector carrying the hTERT (human telomerase reverse transcriptase) catalytic subunit gene, and investigated their differentiation potential. Transfected hUCMSCs overexpressed the hTERT gene and up-regulated their telomerase activity. The transfected hUCMSCs maintained their typical morphology and MSC-specific markers, and vigorously proliferated, undergoing more than 100 PDs (population doublings) to date. Following incubation with hepatogenic agents, the transfected hUCMSCs differentiated into hepatocyte-like cells, and expressed hepatic markers, such as albumin, AFP (α-fetoprotein) and CK-18 (cytokeratin-18). Transfected hUCMSCs showed no transformation into tumours in nude mice. In conclusion, telomerization of hUCMSCs by hTERT overexpression extends their replicative lifespan without influencing their hepatogenic differentiation potential. This offers opportunities for obtaining sufficient quantities of cells for liver-directed therapies.

[Impact of body mass index on blood pressures in a college student population].

OBJECTIVE: To explore the morbidity of hypertension and prehypertension and analyze the association between body mass index (BMI) and blood pressure in Chinese college students. METHODS: This epidemiological study involved 490 Chinese college students (aged 15-25 years, mean 18.9∓1.2 years), and their body height, body weight, blood pressure and heart rate were recorded. The BMI was divided into four groups according to Chinese population standard classification. RESULTS: Most students were southern Han Chinese. The incidences of overweight plus obesity and obesity were 12.9% and 4.1%, respectively. The morbidity of hypertension was 0.8% in this student population. The incidences of hypertension and pre-hypertension were higher in high BMI groups than those in low BMI groups. The systolic blood pressures were significantly higher in over-weight and obese groups than in the normal BMI and lean groups. BMI was positively correlated to systolic and diastolic blood pressures in this population. CONCLUSION: High BMI is a predictor of elevated blood pressure in adolescent students.

[Test-retest reliability of Mandarin monosyllable lists: a multi-center study in Chinese dialectal regions].

OBJECTIVE: To develop 22 Chinese Mandarin monosyllable lists with good psychometrical equivalence. This study was to evaluate the test-retest reliability of these lists when it was used in speech recognition test in normal hearing dialectal speakers. METHODS: Seven cities including Dalian, Shanghai, Hangzhou, Wuhan, Guangzhou, Fuzhou and Xiamen were selected as testing centers which contain 6 typical Chinese dialectal regions including north of China, East of China, north of Fujian, south of Fujian, Guangdong and mid-south of China. At each center, 22 local normal hearing people were selected to join this study. Every participant was tested by each recognition test of all 22 lists twice in two sessions and same test order respectively. The second run of testing was carried out within 10 days-1 month since first run of testing. RESULTS: There was a significant correlations between scores obtained at the two sessions (r = 0.682, P < 0.01). Paired student-t test had shown that a gross score of all dialectal participants was significantly higher than that of initial test to retest (P < 0.01). The mean increment of score was (2.7 +/- 10.1)%. A significant difference of test-retest score in 7 sites was 19.8% and it was equal to 5 test items. A one way ANOVA analysis had indicated that there were statistically significant difference between the score improvement of 7 test sites (P < 0.01). Another analysis had shown that there was no significant correlation between test-retest score improvement and intra-session intervals (P = 0.947). CONCLUSIONS: Mandarin monosyllabic recognition test seems to be more stable, and the present study has indicated a systematic differences in Chinese Mandarin monosyllable recognition scores between test and retest. Monosyllable recognition test is not susceptible to memory effect. Pearson's correction analysis is not suitable to evaluation for test-retest reliability.