Effect of the traditional Chinese herb Helminthostachys zeylanica on postsurgical recovery in patients with ankle fracture: A double-blinded randomized controlled clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Helminthostachys zeylanica (HZ), which is also called "Dao-Di-U-Gon" in Taiwan, has anti-inflammatory and antiedema effects and is commonly used to treat edema in patients with fractures. The ugonin K component of HZ can induce osteogenesis and promote bone mineralization, its therapeutic effect, however, its therapeutic effect remains unclear. Therefore, the purpose of the present study was to investigate the effect of HZ on functional recovery in patients with ankle fractures requiring surgical treatment. METHODS: A double-blinded, randomized, controlled study was conducted. A total of 45 patients with ankle fractures requiring surgical treatment were assigned to either the control group (n = 23 patients), which received the oral administration of HZ placebo 1.0 g t.i.d. for 42 days continuously, or to the treatment group (22 patients), which received HZ for 42 days. RESULTS: The serum amino-terminal propeptide of type 1 procollagen (PINP) levels were similar in the first assessment (V1) between the control (45.90 ± 16.31 ng/mL) and treatment groups (52.61 ± 21.02 ng/mL; p = 0.240); the differences in PINP level between the third assessment (V3) and V1 were greater in the treatment group (35.84 ± 24.56 ng/mL) than in the control group (16.34 ± 11.97 ng/mL; p = 0.002). Radiographic healing time (RHT) was 9.09 ± 1.15 weeks in the treatment group, which was shorter than the 9.91 ± 0.79 weeks (p = 0.012) in the control group. CONCLUSION: Oral administration of HZ for 42 days can increase serum PINP level and reduce the RHT. Therefore, HZ can be used to treat patients with ankle fractures requiring surgical treatment. However, a larger sample size is needed in future studies.

An Intermediate Concentration of Calcium with Antioxidant Supplement in Culture Medium Enhances Proliferation and Decreases the Aging of Bone Marrow Mesenchymal Stem Cells.

Human bone marrow stem cells (HBMSCs) are isolated from the bone marrow. Stem cells can self-renew and differentiate into various types of cells. They are able to regenerate kinds of tissue that are potentially used for tissue engineering. To maintain and expand these cells under culture conditions is difficult-they are easily triggered for differentiation or death. In this study, we describe a new culture formula to culture isolated HBMSCs. This new formula was modified from NCDB 153, a medium with low calcium, supplied with 5% FBS, extra growth factor added to it, and supplemented with N-acetyl-L-cysteine and L-ascorbic acid-2-phosphate to maintain the cells in a steady stage. The cells retain these characteristics as primarily isolated HBMSCs. Moreover, our new formula keeps HBMSCs with high proliferation rate and multiple linage differentiation ability, such as osteoblastogenesis, chondrogenesis, and adipogenesis. It also retains HBMSCs with stable chromosome, DNA, telomere length, and telomerase activity, even after long-term culture. Senescence can be minimized under this new formulation and carcinogenesis of stem cells can also be prevented. These modifications greatly enhance the survival rate, growth rate, and basal characteristics of isolated HBMSCs, which will be very helpful in stem cell research.

Green Tea Catechin (-)-Epigallocatechin-3-Gallate (EGCG) Facilitates Fracture Healing.

Green tea drinking can ameliorate postmenopausal osteoporosis by increasing the bone mineral density. (-)-Epigallocatechin-3-gallate (EGCG), the abundant and active compound of tea catechin, was proven to be able to reduce bone loss and ameliorate microarchitecture in female ovariectomized rats. EGCG can also enhance the osteogenic differentiation of murine bone marrow mesenchymal stem cells and inhibit the osteoclastogenesis in RAW264.7 cells by modulation of the receptor activator of nuclear factor-kB (RANK)/RANK ligand (RANKL)/osteoprotegrin (OPG) (RANK/RANKL/OPG) pathway. Our previous study also found that EGCG can promote bone defect healing in the distal femur partially via bone morphogenetic protein-2 (BMP-2). Considering the osteoinduction property of BMP-2, we hypothesized that EGCG could accelerate the bone healing process with an increased expression of BMP-2. In this manuscript, we studied whether the local use of EGCG can facilitate tibial fracture healing. Fifty-six 4-month-old rats were randomly assigned to two groups after being weight-matched: a control group with vehicle treatment (Ctrl) and a study group with 10 µmol/L, 40 µL, EGCG treatment (EGCG). Two days after the operation, the rats were treated daily with EGCG or vehicle by percutaneous local injection for 2 weeks. The application of EGCG enhanced callus formation by increasing the bone volume and subsequently improved the mechanical properties of the tibial bone, including the maximal load, break load, stiffness, and Young's modulus. The results of the histology and BMP-2 immunohistochemistry staining showed that EGCG treatment accelerated the bone matrix formation and produced a stronger expression of BMP-2. Taken together, this study for the first time demonstrated that local treatment of EGCG can accelerate the fracture healing process at least partly via BMP-2.

(-)-Epigallocatechin-3-gallate (EGCG) enhances healing of femoral bone defect.

BACKGROUND: Previously, we found that (-)-epigallocatechin-3-gallate (EGCG) enhanced osteogenic differentiation of murine bone marrow mesenchymal stem cells by increasing the mRNA expression of osteogenesis-related genes, alkaline phosphatase activity and eventually mineralization. We further found EGCG supplementation preserved bone mass and microarchitecture in female rats during estrogen deficiency in the proximal tibia and lumbar spine at least in part by increasing bone morphogenetic protein-2 (BMP2). BMP2 can enhance de novo bone formation. PURPOSE: In this study, we evaluate the effect of local EGCG application in de novo bone formation in bone defect healing. METHODS: Twenty-four rats aged 4 months were weight-matched and randomly allocated to 2 groups: defect control with vehicle treatment (control) and defect with 10 µM EGCG treatment (EGCG). Daily vehicle and EGCG were applied locally by percutaneous local injection 2 days after defect creation for 2 weeks. Four weeks after treatment, animals were sacrificed for micro-computed tomography (µ-CT) and biomechanical analysis. RESULTS: Local EGCG at femoral defect can enhance de novo bone formation by increasing bone volume and subsequently improve mechanical properties including max load, break point, stiffness, area under the max load curve, area under the break point curve and ultimate stress. CONCLUSIONS: Local EGCG may enhance bone defect healing via at least partly by the de novo bone formation of BMP-2.

Exercise training ameliorates glucosamine-induced insulin resistance in ovariectomized rats.

OBJECTIVE: Glucosamine (GlcN), which has been reported to induce insulin resistance (IR), is a popular nutritional supplement used to treat osteoarthritis in menopausal women. We previously demonstrated that GlcN treatment caused IR in ovariectomized rats by reducing the expression of glucose transport protein subtype 4 (GLUT-4) in skeletal muscle. In the present study, we hypothesized that endurance exercise training can reverse GlcN-induced IR. METHODS: Fifty female rats were randomly divided into five groups with 10 rats in each group: (1) sham-operated group; (2) sham-operated group with GlcN treatment for 14 days; (3) ovariectomy (OVX) group; (4) OVX with GlcN treatment; and (5) OVX with GlcN treatment followed by exercise training (running program) for 8 weeks. RESULTS: Fasting plasma glucose increased in the OVX + GlcN group, and fasting plasma insulin and the homeostasis model assessment-insulin resistance (HOMA-IR) were significantly higher only in this group. After the rats received exercise training for 8 weeks, no increase in the fasting plasma glucose, insulin, or HOMA-IR was observed. In an intraperitoneal glucose tolerance test, the plasma glucose, plasma insulin, HOMA-IR, and glucose-insulin index were significantly elevated only in the OVX with GlcN treatment group. However, the plasma glucose, plasma insulin, HOMA-IR, and glucose-insulin index decreased after exercise training for 8 weeks, implying that GlcN-induced IR in OVX rats could be reversed through exercise. A histological analysis revealed that exercise training can reduce islet hypertrophy and maintain GLUT-4 in skeletal muscle. CONCLUSIONS: Exercise training can alleviate IR in OVX rats treated with GlcN. Islet hyperplasia was subsequently prevented. Preserving GLUT-4 expression may be one of the mechanisms by which exercise prevents IR.

TCM tongue diagnosis index of early-stage breast cancer.

OBJECTIVES: This paper investigates discriminating tongue features to distinguish between early stage breast cancer (BC) patients and non-breast cancer individuals through non-invasive traditional Chinese medicine (TCM) tongue diagnosis. DESIGN: The tongue features for 67 patients with 0 and 1 stages of BC, and 70 non-breast cancer individuals are extracted by the automatic tongue diagnosis system (ATDS). A total of nine tongue features, namely, tongue color, tongue quality, tongue fissure, tongue fur, red dot, ecchymosis, tooth mark, saliva, and tongue shape are identified for each tongue. Features extracted are further sub-divided according to the areas located, i.e., spleen-stomach, liver-gall-left, liver-gall-right, kidney, and heart-lung areas. This study focuses on deriving significant tongue features (p<0.05) to discriminate early-stage BC patients from non-breast cancer individuals. RESULTS: The Mann-Whitney test shows that the amount of tongue fur (p=0.024), maximum covering area of tongue fur (p=0.009), thin tongue fur (p=0.009), the average area of red dot (p=0.049), the maximum area of red dot (p=0.009), red dot in the spleen-stomach area (p=0.000), and red dot in the heart-lung area (p=0.000) demonstrate significant differences. The data collected are further classified into two groups. The training group consists of 57 early-stage BC patients and 60 non-breast cancer individuals, while the testing group is composed of 10 early-stage BC patients and 10 non-breast cancer individuals. The logistic regression by utilizing these 7 tongue features with significant differences in Mann-Whitney test as factors is performed. In order to reduce the number of tongue features employed in prediction, tongue features with the least amount of significant difference, namely, maximum area of red dot and average area of red dot, are removed progressively. The tongue features of the testing group are employed in the aforementioned three models to test the power of significant tongue features identified in predicting early-stage BC. An accuracy of 80%, 80% and 90% is reached on non-breast cancer individuals by applying the 7, 6 and 5 significant tongue features obtained through Mann-Whitney test, respectively, while 60%, 60% and 50% is reached on the corresponding early-stage BC patients. CONCLUSION: The TCM tongue diagnosis can serve as a preliminary screening procedure in the early detection of BC in light of its simple and non-invasive nature, followed by other more accurate testing process. To the best of our knowledge, this is the first attempt in applying non-invasive TCM tongue diagnosis to the discrimination of early-stage BC patients and non-breast cancer individuals.

Education of Chinese medicine tongue diagnosis by automatic tongue diagnosis system.

OBJECTIVE: To investigate the efficacy of applying automatic tongue diagnosis system (ATDS) to the training of novice Chinese medicine (CM) doctors in the tongue diagnosis. METHOD: A two-round experiment composed of pre- and after-training tests was performed. Two groups of CM doctors were formed by experienced doctors (ED) and novice doctors (ND). The intra- and inter-observer agreements for the ED and ND groups, and the inter-observer agreements between ATDS and ED, and ATDS and ND were derived first. After a 1-month training of the ND, the intra- and inter-observer agreements of the ND group, and the inter-observer agreement between ATDS and ND were obtained. RESULT: The kappa value of agreement data for ND after training was significantly higher than ND before training with a level of P<0.05, indicating an obvious improvement of intra-observer agreement for ND after training with ATDS. The inter-observer agreements for the ED group, and ND group after training were significantly higher (P<0.05) than the ND group before training. In contrast, no significance is observed between the ED group and the ND group after training, indicating that physicians in the ND group achieve an inter-observer agreement to the same level as that of ED after 1-month training. CONCLUSION: This finding validates the utilization of ATDS as a mean of education in forging consensus during the learning process of tongue diagnosis.

Baicalein inhibits HMGB1 release and MMP-2/-9 expression in lipopolysaccharide-induced cardiac hypertrophy.

Myocardial dysfunction, a common complication after sepsis, significantly contributes to the death of patients with septic shock. In the search for potentially effective drugs to decrease mortality from sepsis, we investigated the cardioprotective effects of baicalein, a flavonoid present in the root of Scutellaria baicalensis, on lipopolysaccharide (LPS)-induced pro-inflammatory cytokine production and matrix metalloproteinase-2 and -9 (MMP-2/-9) expression. We found that baicalein significantly attenuated LPS-induced cardiac hypertrophy and counteracted reactive oxygen species (ROS) generation in neonatal rat cardiomyocytes. In addition, pretreatment with baicalein inhibited LPS-induced early (e.g., tumor necrosis factor-α (TNF-α) and interleukin-6) and late (e.g., high mobility group box 1 (HMGB1) pro-inflammatory cytokine release, inducible nitric oxide synthase (iNOS) expression and NO production. Finally, baicalein also significantly down-regulated the expression of MMP-2/-9 and attenuated HMGB1 translocation from the nucleus to the cytoplasm. These results suggest that baicalein can protect cardiomyocytes from LPS-induced cardiac injury via the inhibition of ROS and inflammatory cytokine production. These cardioprotective effects are possibly mediated through the inhibition of the HMGB1 and MMP-2/-9 signaling pathways.

Tongue diagnosis of traditional Chinese medicine for rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic, systemic autoimmune disease with unknown aetiology that causes the immune system to attack the joints (synoviums), leading to chronic inflammation. According to the traditional Chinese medicine (TCM), RA falls into the category of Impediment disease ("Bi" syndrome), that is, poor circulation of qi and blood (stasis). Tongue diagnosis is an important method of TCM to detect blood stasis. In this study, 74 RA patients, meeting the pre-set criteria, were recruited via rheumatology outpatient clinic and examined by experienced rheumatology physicians. Two images-one of the tongue and the other, sublingual vessels-of the same patient were taken by a Canon digital camera in a darkroom with uniform lighting conditions. Relevant features of the tongue were extracted by utilising image processing techniques. Every tongue was classified into corresponding patterns based on the features identified. The subjects included 62 females and 12 males with an average age of 49.86 ± 13.81 years old, an average morbidity period of 4.56 ± 3.92 years, an average rheumatoid factor (RF) of 225.3 ± 373.8 IU/mL and an average erythrocyte sedimentation rate of (ESR) 40.9 ± 31.9 m/hr. According to our study, 86% of the patients with RA have tongues with sublingual vessels with a width of more than 2.7 mm, a length of more than 3/5 from tongue tipto sublingual caruncle, or a count of sublingual vessels more than 2. Moreover, since RA index is highly correlated with blood stasis in TCM, a logistic regression is conducted to predict the probability of presence of RA using RF and ESR as explanatory variables. Also, the logistic regression analysis of RA with respect to the conventional tongue diagnosis criteria was performed. Based on the aforementioned studies, we concluded that tongue diagnosis is helpful in detecting blood stasis of RA.

On the hazard caused by the heat of acupuncture needles in warm needling (wen zhen).

Due to its simplicity and convenience, acupuncture has become popular as a complementary therapy. In this Chinese medicine, doctors have to find the traditional meridian acupuncture points before puncturing the needles into them. Moxibustion ( Ài Jiǔ) is also an important part of the acupuncture remedy. Treatment by acupuncture can be classified roughly into two types - direct moxibustion and indirect moxibustion. Warm-needling acupuncture ( Wen Zhen Jiǔ) is classified under the method of indirect moxibustion. In the present study, 10 standard stainless steel acupuncture needles with 10 pieces of cylinder-shaped moxa cone ( Ài Zhù) as the heat source of warm needles were used. In order to prevent the practitioners from getting burns, it is necessary to study the temperature changes in some designated parts of the needles. Two sizes, 0.6 g and 1.0 g, of moxa cones were used for comparison of the measured temperatures. The needles are typically divided into two parts - the handle part and the needle body. In our experiment, the temperatures of WNA at different parts of the needles were measured. The larger the size of moxa cone is, the longer is the burning time. Based on the observations we suggest that when 0.6 g moxa is used, the physicians should better pick out the needles around 9 min after ignition; however, while using the 1 g moxa, it might be safer to pick out the needles around 13 min after ignition.

Breast cancer index: a perspective on tongue diagnosis in traditional chinese medicine.

Breast cancer (BC) ranks second in the cancer fatality rate among females worldwide. Mammogram, ultrasound, magnetic resonance imaging (MRI), blood testing, and fine needle aspiration biopsy are usually applied to discriminate BC patients from normal persons. False-negative results, undetectable calcifications, movement-incurred blurry image, infection, and sampling error are commonly associated with these traditional means of diagnosis. Traditional Chinese medicine (TCM) covers a broad range of medical practices sharing common theoretical concepts. Tongue diagnosis plays an important role in TCM. Organ conditions, properties, and variation of pathogens can be revealed through observation of tongue. In light of this observation, this paper investigates discriminating tongue features to distinguish between BC patients and normal people, and establishes differentiating index to facilitate the non-invasive detection of BC. The tongue features for 60 BC patients and 70 normal persons were extracted by the Automatic Tongue Diagnosis System (ATDS). The Mann-Whitney test showed that the amount of tongue fur (P = 0.007), tongue fur in the spleen-stomach area, maximum covering area of tongue fur, thin tongue fur, the number of tooth marks, the number of red dots, red dot in the spleen-stomach area, red dot in the liver-gall-left area, red dot in the liver-gall-right area, and red dot in the heart-lung area demonstrated significant differences (P < 0.05). The tongue features of the testing group were employed to test the power of significant tongue features identified in predicting BC. An accuracy of 80% was reached by applying the seven significant tongue features obtained through Mann-Whitney test. To the best of our knowledge, this is the first attempt in applying TCM tongue diagnosis to the discrimination of BC patients and normal persons.

Visual agreement analyses of traditional chinese medicine: a multiple-dimensional scaling approach.

The study of TCM agreement in terms of a powerful statistical tool becomes critical in providing objective evaluations. Several previous studies have conducted on the issue of consistency of TCM, and the results have indicated that agreements are low. Traditional agreement measures only provide a single value which is not sufficient to justify if the agreement among several raters is strong or not. In light of this observation, a novel visual agreement analysis for TCM via multiple dimensional scaling (MDS) is proposed in this study. If there are clusters present in the raters in a latent manner, MDS can prove itself as an effective distinguisher. In this study, a group of doctors, consisting of 11 experienced TCM practitioners having clinical experience ranging from 3 to 15 years with a mean of 5.5 years from the Chinese Medicine Department at Changhua Christian Hospital (CCH) in Taiwan were asked to diagnose a total of fifteen tongue images, the Eight Principles derived from the TCM theorem. The results of statistical analysis show that, if there are clusters present in the raters in a latent manner, MDS can prove itself as an effective distinguisher.

The Study on the Agreement between Automatic Tongue Diagnosis System and Traditional Chinese Medicine Practitioners.

Tongue diagnosis is an important practice in traditional Chinese medicine (TCM) for diagnosing diseases before determining proper means of treatments. Traditionally, it depends solely on personal knowledge and experience of the practitioner, thereby being criticized as lacking of objectivity. Currently, no research regarding intra- and inter-agreements of automatic tongue diagnosis system (ATDS) and TCM doctors has been conducted. In this study, the ATDS is developed to extract a variety of tongue features and provide practitioners with objective information to assist diagnoses. To evaluate the ATDS clinical stability, 2 sets of tongue images taken 1 hour apart from 20 patients with possible variations in lighting and extruding tongue, are employed to investigate intra-agreement of the ATDS, intra-agreement of the TCM doctors, and the inter-agreement between the ATDS and TCM doctors. The ATDS is shown to be more consistent with significantly higher intra-agreement than the TCM doctors (kappa value: 0.93 ± 0.06 versus 0.64 ± 0.13) with P < 0.001 (Student's t-test). Inter-agreements between the ATDS and TCM doctors, as well as among the TCM doctors are both moderate. The high agreement of the ATDS can provide objective and reliable tongue features to facilitate doctor in making effective observation and diagnosis of specific diseases.

Analysis of agreement on traditional chinese medical diagnostics for many practitioners.

In Traditional Chinese Medicine (TCM) diagnostics, it is an important issue to study the degree of agreement among several distinct practitioners. In order to study the reliability of TCM diagnostics, we have to design an experiment to simultaneously deal with both of the cases when the data is ordinal and when there are many TCM practitioners. In this study, we consider a reliability measure called "Krippendorff's alpha" to investigate the agreement of tongue diagnostics in TCM. Besides, since it is not easy to obtain a large data set with patients rated simultaneously by many TCM practitioners, we use the renowned "bootstrapping" to obtain a 95% confidence interval for the Krippendorff's alpha. The estimated Krippendorff's alpha for the agreement among ten physicians that discerned fifteen randomly chosen patients is 0.7343, and the 95% bootstrapping confidence interval for the true alpha coefficient is [0.6570, 0.7349]. The data was collected and analyzed at the Department of Traditional Chinese Medicine, Changhua Christian Hospital (CCH) in Taiwan.