[Mechanism of electroacupuncture penetration needling for relieving synovial inflammation of knee osteoarthritis through TLR4/MyD88/NF-κB signal pathway].

OBJECTIVE: To observe the effects of electroacupuncture (EA) penetration needling on Toll-like receptors 4/myeloid differentiation factor 88/nuclear factor-kappa B (TLR4/MyD88/NF-κB) signaling pathway in rat synovium and the serum-related inflammatory factors, so as to explore the mechanism of EA penetration needling on synovial inflammation in rats with knee osteoarthritis (KOA). METHODS: SD male rats were randomly divided into sham-operation group, model group, EA+penetration needling group, and conventional EA group, with 16 rats in each group. The rats model was prepared by anterior cruciate ligment transection and these rats were forced to exercise for 8 weeks after operation. After successful modeling, in the EA+penetration needling group, the needles were inserted at "Dubi" (ST35) "Neixiyan" (EX-LE4), and at "Xuehai"(SP10) "Liangqiu"(ST34) on the right hind limb, towards each other, 5-8 mm in depth, respectively. In the conventional EA group, the needles were inserted at ST35 and EX-LE4 on the right hind limb, obliquely, at 30° angle to the skin, 3-5 mm in depth; and were inserted at SP10 and ST34 on the right hind limb perpendicularly, 3-5 mm in depth. In these two groups, electric stimulation was operated with dense-disperse wave, 2 Hz/10 Hz in frequency and 0.5-1.5 mA in intensity, retained for 20 min in each treatment. The treatment was given once daily, 10 days as 1 course of treatment, and 2 courses were required at the interval of 2 days. After the intervention, the knee joint effusion was observed by musculoskeletal ultrasound; the contents of IL-1ß, IL-6 and TNF-α in serum were determined by ELISA; the morphological changes in the synovium were observed after H.E. staining; the positive expression of NF-κB p65 in the synovial membrane was detected by immunohistochemical method; the expression levels of TLR4, MyD88, TRAF-6 and NF-κB p65 proteins in the synovial membrane were determined by Western blot. RESULTS: Compared with the sham-operation group, in the model group, the knee joint effusion was obviously increased, the synovial lining cells were distributed irregularly, the cells were disarranged, the pannus was formed largely, and a great number of the inflammatory cells were infiltrated; the contents of serum IL-1ß, IL-6 and TNF-α, the positive expression of NF-κB p65, the protein expression levels of TLR4, MyD88, TRAF-6 and NF-κB p65 in the synovial tissue were increased (P<0.05). Compared with the model group, the knee joint effusion was reduced, the synovial lining cells were proliferated, a small number of the inflammatory cells were infiltrated, and the pannus was formed lightly; the contents of serum IL-1ß, IL-6 and TNF-α, the positive expression of NF-κB p65, the protein expression levels of TLR4, MyD88, TRAF-6 and NF-κB p65 in the synovial tissue were lower (P<0.05) in the EA+penetration needling group and the conventional EA group. In the conventional EA group, the knee joint effusion was increased, the synovial lining cells were proliferated, the inflammatory cells were infiltrated largely, and the pannus was formed increasingly; the contents of serum IL-1ß, IL-6 and TNF-α, and the protein expression levels of TLR4, MyD88 and NF-κB p65 in the synovial tissue were increased when compared with the EA+penetration needling group (P<0.05). CONCLUSION: The EA+penetration needling can significantly relieve the synovial inflammatory reaction and the knee joint effusion in KOA rats. The mechanism is probably related to down-regulating the downstream inflammatory cascade through inhibiting the transduction of TLR4/MyD88/NF-κB signaling pathway.

Valproic acid attenuates global cerebral ischemia/reperfusion injury in gerbils via anti-pyroptosis pathways.

Ischemic stroke is the third most common cause of death and the leading cause of disability worldwide in adults. The antiepileptic drug valproic acid (VPA) was reported to protect cerebral ischemia/reperfusion injury. However, the action mechanism of VPA in cerebral ischemia/reperfusion injury has not been fully understood. We explored the action mechanism of VPA in vivo and in vitro. Gerbils were subjected to transient global cerebral ischemic-reperfusion injury, and hippocampal neuron injury was treated with oxygen-glucose deprivation in vitro. Morris water maze test was performed to evaluate the cognitive dysfunction. Histopathological examinations and western blot were performed to evaluate the pyroptosis of neurons. The results showed that VPA attenuated the cognitive dysfunction, pyroptosis of the gerbils suffer from ischemic-reperfusion injury and decreased hippocampal neurons pyroptosis induced by oxygen-glucose deprivation in vitro. In addition, western blot and real-time PCR analysis revealed that VPA modulated the protein expression of apoptosis repressor with caspase recruitment domain (ARC), caspase-1 and IL-1ß/IL-18. Our results suggested that VPA alleviated ischemic/reperfusion injury-mediated neuronal impairment by anti-pyroptotic effects.

Boronate-functionalized hydrogel as a novel biosensing interface for the glycated hemoglobin A1c (HbA1c) based on the competitive binding with signaling glycoprotein.

According to recent increases in public healthcare costs associated with diabetes mellitus, the development of new glycemic monitoring techniques based on the biosensing of glycated hemoglobin A1c (HbA1c), a promising long-term glycemic biomarker, has become a major challenge. In the development of HbA1c biosensors for point-of-care applications, the selection of an effective biorecognition layer that provides a high reaction yield and specificity toward HbA1c is regarded as the most significant issue. To address this, we developed a novel HbA1c biosensing interfacial material by the integration of boronate hydrogel with glass fiber membrane. In the present study, a new boronate-functionalized hydrogel was designed and spatio-selectively photopolymerized on a hydrophilic glass fiber membrane by using N-hydroxyethyl acrylamide, 3-(acrylamido)phenylboronic acid, and bis(N,N'-methylene-bis-acrylamide). Using this approach, the boronic acid group, which specifically recognizes the cis-diol residue of glucose on the HbA1c molecule, can be three-dimensionally coated on the surface of the glass fiber network with a high density. Because this network structure of boronate hydrogel-grafted fibers enables capillary-driven fluid control, facile HbA1c biosensing in a lateral flow assay concept could be accomplished. On the proposed HbA1c biosensing interface, various concentrations of HbA1c (5-15%) in blood-originated samples were sensitively measured by a colorimetric assay using horseradish peroxidase, a glycoenzyme can generate chromogenic signal after the competitive binding against HbA1c to the boronic acid residues. Based on the demonstrated advantages of boronate hydrogel-modified membrane including high analytical performance, easy operation, and cost-effectiveness, we expect that the proposed biorecognition interfacial material can be applied not only to point-of-care HbA1c biosensors, but also to the quantitative analysis of other glycoprotein biomarkers.

[A cohort study on the relationship between cerebrovascular hemodynamic changing and risk of stroke].

OBJECTIVE: To study the role of cerebrovascular hemodynamic indexes (CVHI) changing in stroke and to provide reference for stroke prevention and risk factor study. METHODS: From 2003 to 2004, participants aged 40 years and above in two communities in Fengxian district were recruited by cluster sampling. Risk factors of stroke and CVHI were investigated and checked during baseline investigation. A total of 10 565 individuals completed the survey and met the inclusion criterion. After baseline investigation, the cohort was followed up for stroke occurrence. Relative risk (RR) of CVHI and common risk factors were estimated by cohort study design. RESULTS: Age of the cohort was (56.2 ± 11.4) years. 4444 (42.1%) were males and 6121 (57.9%) were females. Total follow-up duration was 67 885.7 person-years. A total of 195 stroke cases occurred and incidence density of stroke was 287.2 per 100 000 person-years. Stroke incidence in exposure groups of hypertension, heart disease and alcohol drinking was 3.47% (108/3118), 2.96% (21/710) and 2.50% (47/1882), respectively. The incidence in corresponding non-exposure group was 1.17% (87/7448), 1.77% (174/9855) and 1.70% (148/8683) respectively. There was significant difference between 2 groups (χ(2) value was 62.72, 4.56 and 4.94, respectively, P < 0.05). Stroke incidence in CVHI score < 25, 25 - 49, 50 - 74 and ≥ 75 groups was 9.12% (59/647), 5.68% (44/775), 2.52% (39/1545) and 0.72% (53/7403)(χ(2)trend = 273.57, P < 0.05), respectively. Incidence of stroke in 40 - 49, 50 - 59, 60 - 69, ≥ 70 years age group was 0.22% (8/3565), 1.28% (43/3357), 2.71% (50/1848) and 5.88% (94/1600) (χ(2)trend = 181.48, P < 0.05), respectively. Multiple Cox regression analysis indicated that RR (95%CI) value of hypertension and cigarette smoking was 1.40(1.02 - 1.92) and 1.59(1.19 - 2.12), respectively when comparing with non-exposure group. RR (95%CI) value in CVHI score < 25, 25 - 49 and 50 - 74 points group were 6.15 (4.08 - 9.26), 4.55 (2.98 - 6.96) and 2.68 (1.75 - 4.09), respectively when comparing with the score ≥ 75 points group. RR (95%CI) value in age 50 - 59, 60 - 69 and ≥ 70 years group was 4.61 (2.16 - 9.82), 7.81 (3.67 - 16.60) and 13.49(6.44 - 28.24), respectively when comparing with below 40 years group. CONCLUSION: CVHI score is the strong independent predictive factor and hypertension, cigarette smoking and age are the independent risk factors of stroke.

Bioelectrocatalytic detection of glycated hemoglobin (HbA1c) based on the competitive binding of target and signaling glycoproteins to a boronate-modified surface.

We developed an electrochemical glycated hemoglobin (HbA(1c)) biosensor for diagnosing diabetes in whole human blood based on the competitive binding reaction of glycated proteins. Until now, no studies have reported a simple and accurate electrochemical biosensor for the quantification of HbA(1c) in whole blood. This is because it is very difficult to correctly distinguish HbA(1c) from large amounts of hemoglobin and other components in whole blood. To detect glycated hemoglobin, we used electrodes modified with boronic acid, which forms a covalent bond between its diol group and the cis-diol group of the carbohydrate moiety of glycated proteins. For accurate HbA(1c) biosensing, we first removed blood components (except for hemoglobin) such as glycated proteins and blood glucose as they interfere with the boronate-based HbA(1c) competition analysis by reacting with the boronate-modified surface via a cis-diol interaction. After hemoglobin separation, target HbA(1c) and GOx at a predetermined concentration were reacted through a competition onto the boronate-modified electrode, allowing HbA(1c) to be detected linearly within a range of 4.5-15% of the separated hemoglobin sample (HbA(1c)/total hemoglobin). This range covers the required clinical reference range of diabetes mellitus. Hence, the proposed method can be used for measuring %HbA(1c) in whole human blood, and can also be applied to measuring the concentration of various glycated proteins that contain peripheral sugar groups.

[Study on over-weight, obesity and the exposure to stroke risk factors among 40 year olds and over in a community-based population].

OBJECTIVE: To describe the distribution of over weight and obesity to discover the level of exposure on risk factors of stroke among people aged 40 and over, in a community-based population. METHODS: From 2003 to 2004, people aged > or = 40 years in a community of Fengxian district in Shanghai were selected by cluster sampling. Out of 11,791 individuals who were selected, 10,565 met the inclusion criteria and responded to the investigation. By face to face interview, a cross-sectional survey was carried out, using a questionnaire for risk factors of stroke. Height, weight and blood pressures were measured and cerebrovascular hemodynamic parameters (CVHP) were checked. Age and gender distribution of over weight and obesity in the population were described. Using 60 year as cut-off point, participants were grouped into three: normal, over weight and obesity by body mass index. Level of stroke risk factor exposure between groups was compared and logistic regression model was used for multiple analyses. RESULTS: Proportions of over weight and obesity were 28.5 percent and 4.1 percent in male and 26.3 percent and 4.2 percent in female (P = 0.045). Systolic and diastolic blood pressure in over weight group were (132.5 +/- 19.4) mm Hg and (83.9 +/- 10.5) mm Hg (1 mm Hg = 0.133 kPa), which were higher than that in normal weight group and lower than that in obese group (P < 0.05). Exposure rate of heart disease, family history of stroke in < 60 year old group and diabetes in > or = 60 year group increased along with the increase of weight. Exposure rate of hypertension, abnormality of CVHP score in both age groups were also increased with the increase of weight. Data from multiple logistic regression indicated that hypertension, family history of stroke and heart disease, CVHP score below 75 points, sex and age were independent factors of over weight and obese. CONCLUSION: The prevalence of over weight or obesity in a community-based population among aged 40 years or over was around 30 percent. The overall exposure rate of stroke risk factors were increasing along with the increase of weight, especially for those in the middle age.

[Baseline study and analysis on a stroke risk factor-related cohort in Fengxian county of Shanghai].

OBJECTIVE: To explore the exposed level of stroke risk factors in Fengxian county of Shanghai and the relationships between risk factors and stroke. METHODS: Community based populations including 10,565 individuals aged 40 years old and over were cluster sampled from Fengxian county in 2003 or 2004. Status of exposure on common risk factors such as hypertension, cardiovascular disease, diabetes, family history of stroke, cigarette smoking, alcohol intake were gathered while cerebral vascular hemodynamic index (CVHI) was examined, with CVHI score below 75 points as abnormal. During 2003 to 2006, 78 stroke cases occurred. The relationships between risk factors and stroke were analyzed by univariate and Cox proportional hazards regression models. RESULTS: Rates of exposure regarding hypertension, cardiovascular disease, diabetes, family history of stroke, obesity (BMI > or = 28 kg/m2), smoking, drinking and CVHI score abnormal were 21.14%, 6.72%, 1.88%, 5.63%, 4.17%, 34.96%, 17.81% and 29.43%, respectively. Data from Univariate analysis indicated that relative risk (95% CI) of above-mentioned risk factors were 2.76 (1.76-4.32), 2.19 (1.16-4.14), 1.52 (0.38-6.19), 1.58 (0.69-3.62), 1.24 (0.45-3.38), 1.75 (1.12-2.73), 2.10 (1.30-3.39) and 12.72 (7.02-23.06), respectively. Results from Cox proportional hazards regression models analysis showed that cigarette smoking, CVHI score abnormal were screened into equation. CONCLUSION: Among all the risk factors, rate of hypertension was the highest while hypertension, cardiovascular disease, cigarette smoking, alcohol intake and abnomal CVHI score had remarkable etiological correlations to stroke. Abnormal CVHI score, cigarette smoking seemed to be the independent forecasting factors related to stroke.

[Baseline study on a stroke-related cohort in the rural community of Shanghai].

OBJECTIVE: To present data from a baseline investigation on stroke-related cohort population in rural area of Shanghai. METHODS: A cross-sectional study was carried out in a cluster sampling population aged 40 years and over. General information and data on common risk factors in the population were gathered and cerebral vascular hemodynamic indexes were checked. Hemodynamic score was estimated according to single indexes by unified methods. 5335 persons who had met the inclusion criteria were enlisted in the study. Exposure level of risk factors, prevalence of stroke, and hemodynamic indexes were analyzed and distributional characteristics were described. RESULTS: Exposure rate of hypertension, heart disease, diabetes, family history of hypertension, overweight or obesity in males were 31.74%, 6.09%, 1.16%, 3.22%, 17.64%, 29.68% and were 32.76%, 9.22%, 1.55%, 3.84%, 19.22%, 29.44% in female respectively. Standard prevalence of stroke was 1167.3/100000, which in male was significantly higher than that in female (P < 0.05). The change of cerebral vascular hemodynamic indexes was significantly associated with age. Hemodynamic score in 21.3% of the subjects was below 75 points. CONCLUSIONS: Among population of 40 years old and over in rural areas, hypertension was the principal risk factor regarding the rate of stroke. Prevalence of stroke in males was significantly higher than that in females. Abnormal rate of hemodynamic score was about 20% in this population.

[Effectiveness of Nao'an Capsule on stroke prevention among high risk population in Nanhui, Shanghai].

OBJECTIVE: To evaluate the effectiveness of stroke prevention among high risk population, using Nao'an Capsules. METHODS: Participants were selected from 696,558 residents in Nanhui, using county of Shanghai city. Individuals aged 35 years old and over with at least one risk factor exposure to stroke, received cerebral vascular hemodynamic examination. 18,271 cases meeting the criteria of cerebral vascular hemodynamic indexes accumulative score below 70 points were defined as individuals with high-risk and targets to receive intervention. According to the willingness of the participants, 10,313 cases received Nao'an Capsules for intensive intervention based on general intervention measures compared to 7958 cases only receiving general intervention. After the implementation of intervention, incidence and mortality rates of stroke and the effectiveness of three-year intervention were studied. RESULTS: Incidence of stroke in Nao'an Capsules group was significantly lower than that of the general intervention (P < 0.01) with 53.8% in males and 58.4% in females. The relative risk (RR) in two gender groups were 0.46 (0.33 - 0.64) and 0.39 (0.30 - 0.50) respectively. Multiple logistic regression analysis indicated that the history of hypertension, accumulative score of cerebral vascular hemodynamic indexes, age, gender and Nao'an Capsules intervention were the variables selected into the equation and significantly related to stroke. Among the variables, Nao'an Capsule was the strongest factor with a RR of 0.41. CONCLUSION: Incidence of stroke in Nao'an Capsule intervention group was significantly lower than that in the general intervention group after 3 years of intervention, suggesting that Nao'an Capsule intervention was the strongest factor affecting stroke occurrence in individuals at high-risk.

[The relationship between hypertension and cerebral hemodynamic scores of vascular function accumulative].

OBJECTIVE: To probe into the relationship between hypertension and hemodynamic score of cerebral vascular accumulative function by though studying the history of hypertension and level of blood pressure. METHODS: The database of blood pressure and cerebral vascular hemodynamic indexes (CVHI) were from baseline data and measurement in a cohort. 18,512 participants who met the included criteria were enlisted in the study. The function scores were estimated using uniform methods according to CVHI. Participants were grouped by age, years of having hypertension and level of blood pressure. Differences of the function score between different groups were compared. RESULTS: The function score in hypertension group was significantly lower than that of normal tension group, which decreased by 13.3 points. Within 5 years of hypertension history, the function score decreased as course of hypertension prolonged. The same trend was found between the score and level of both systolic and diastolic blood pressure. When systolic blood pressure elevated to 140 mmHg or diastolic blood pressure elevated to 95 mmHg, the function score was below 75 points. CONCLUSION: There were significant relationship between decrease of the function scores and medical history, course of hypertension as well as level of blood pressure.

[Distribution of cerebral vascular hemodynamic indexes].

OBJECTIVE: To describe the distribution of cerebral vascular hemodynamic indexes (CVHI). METHODS: A number of 25,355 age 35 and over were selected in the Northeast China by cluster sampling. CVHI were checked during baseline survey and were followed to see the occurrence of stroke. Distribution of CVHI among non-stroke population, individuals prior to the onset of stroke and patients with stroke were described. RESULTS: The CVHI accumulative score, V(mean), V(max) and V(min) were dramatically decreasing, but RV, Zcv, WV and DR were significantly increasing as age increased. V(max), RV and CP were significantly higher in males but WV was lower than that of females. The CVHI accumulative score, V(min) and RV were 95.0, 10.23 and 75.8 in non-stroke population, 51.25, 6.71 and 122.72 pre stroke group, and 55.0, 6.78 and 115.89 in patients with stroke respectively. There were significant differences among three groups after controlling of age and sex (P < 0.01). CONCLUSION: Variance of CVHI was closely related to age, and there appeared a significant abnormal of CVHI before and after stroke.

[Principal component analysis and integral methods of cerebral vascular hemodynamic parameters].

OBJECTIVE: To establish a predicting model for stroke according to cerebral vascular hemodynamic indexes and major risk factors of stroke. METHODS: Participants selected from a stroke cohort with 25,355 population in China. The first step was to carry out principal component analysis using CVHI. Logistic regression with principal component and main risk factors of stroke were then served as independent variables and stroke come on as dependent variables. The predictive model was established according to coefficient of regression and probability of each participant was also estimated. Finally, ROC curve was protracted and predictive efficacy was measured. RESULTS: The accumulative contribution rates of four principal components were 58.1%, 79.4%, 88.4% and 94.6% respectively. Seven variables were being selected into the equation with the first to fourth principal component as history of hypertension, age and sex. Area under ROC curve was 0.855 and optimal cut-off point was probability over 0.05. Sensitivity, specificity and accuracy of stroke prediction were 80.7%, 78.5% and 78.5% respectively. CONCLUSION: The model established by principal component and regression could effectively predict the incidence of stroke coming on.

[A cohort study on cerebral vascular hemodynamics accumulative score and risks of stroke].

OBJECTIVE: To estimate relative risk (RR) of cerebral vascular hemodynamics indexes (CVHI) accumulative score as well as some other risk factors of stroke. METHODS: A cohort study was carried out in 25 355 participants aged 35 and over in Northeast of China from 1994 to 2001. In the baseline survey, exposure rates of hypertension, diabetes, family history of stroke and hypertension, etc. were collected and CVHI was examined. CVHI accumulative score was synthesized according to contribution of single index. During the 7-year following up, 228 stroke cases were identified and stroke coming on was defined as the endpoint of observation. Uni-variable and multiple variables analysis were carried out for estimating RR of CVHI and other risk factors of stroke. RESULTS: Uni-variables analysis indicated that RR (95% confidence interval, CI) of Hypertension, heart diseases, diabetes, family history of hypertension and stroke, overweight or obesity, alcohol intake, cigarette smoking and CVHI < 75 scores were 3.23 (2.48 - 4.20), 2.53 (1.92 - 3.33), 2.38 (1.55 - 3.64), 1.32 (1.02 - 1.72), 1.82 (1.37 - 2.41), 1.62 (1.25 - 2.11), 1.48 (1.07 - 2.04), 1.76 (1.34 - 2.31) and 7.30 (5.43 - 9.80) respectively. Cox regression analysis showed that CVHI below 75 points, family history of stroke, cigarette smoking, heart disease, hypertension, age and sex were the factors being selected in equation. There was a significant dosage response between the decrease of CVHI score and the increase of stroke risk. RR of stroke reached 12.55 when CVHI below 75 points and history of hypertension appeared in the same individuals. CONCLUSION: Abnormality of CVHI score seemed to be the most important and independent factor among stroke risk factors. Decrease of CVHI score might serve as a marker of high risk and play important role in stroke, especially coexistence with hypertension.

[Changes of cerebral vascular hemodynamics from normal population to stroke].

OBJECTIVE: To probe into changes of cerebral vascular hemodynamics indexes (CVHI) from normal population to different clinical stage before and after occurring of stroke. METHODS: Participants were selected from 25,355 stroke cohort study population aged 35 years and over in Northeast of China and 55 acute stroke patients were selected from Fuzhou PLA General Hospital. CVHI indexes were checked during baseline investigation or within one week after acute stroke. Participant enlisted in the study were divided into following 5 groups, normal population, high risk population, individuals before stroke, acute stroke patients and convalescence stroke patients. Characteristics of CVHI indexes in different population were analyzed and compared. RESULTS: V(min) of cerebral vascular in previous defined 5 group participants were 11.39 +/- 3.27, 9.66 +/- 3.18, 6.71 +/- 3.30, 4.13 +/- 1.27, 6.78 +/- 3.09, respectively. V(mean) and V(max) were with the same decreasing trends as V(min). However, RV in 5 group participants were 62.35 +/- 21.11, 82.32 +/- 31.16, 122.72 +/- 52.73, 137.46 +/- 49.56 and 115.89 +/- 55.51, respectively. Zcv, WV, DR and CP were also with the same increasing trends as RV. Abnormal rate of CVHI score (< 75 points) from normal population to convalescence stroke patients were 13.3%, 34.7%, 74.1%, 100% and 66.7%, respectively. CONCLUSION: From normal population to clinical stage of stroke, cerebral vascular velocity showed decreasing trends while other indexes, such as RV, Zcv, WV, DR and CP were increasing.

[The reference range of cerebrovascular hemodynamics indexes parameters].

OBJECTIVE: To study the reference range of cerebrovascular hemodynamics indexes (CVHI) in a sampling population. METHODS: Fourteen thousand, four hundred and eighty participants without history of stroke or exposure to risk factors were screened from 26 cities in Northeast China, North China, East China, Northwest China, South China, and Southwest China in the prospective study on stroke intervention. CVHI were checked and 95% confidence intervals were calculated using normal distribution methods, which were grouped by age, sex, area, and check positions. RESULTS: Characteristics of CVHI 95% confidence intervals showed that low limit of carotid blood velocity as V(max), V(min), V(mean) decreased, but upper limit of cerebral RV, Zcv, WV, DR and CP increased with the increase of age. However, Q(mean) and DP did not show significant trends of age difference. Low limit of Q(mean), V(max) and upper limit of RV, DR, CP and DP in males were higher than that in females while upper limit of WV and Zcv in females were higher than that in males. Left carotid low limit of Q(mean), V(mean), V(max), V(min), and upper limit of WV, Zcv and CP were higher than that of the right but upper limit of RV, DR and DP were lower than that of the right. There were no obvious differences between areas. CONCLUSION: Obvious differences of CVHI 95% confidence intervals ranges were found between age groups, sex, and checking positions, respectively, suggesting that the reference ranges of CVHI to be estimated by every subgroup.