Advancing breastfeeding promotion: leveraging integrative natural galactagogues and unveiling their potential roles-Insights from a 19-year Taiwan nationwide registry to address lactation insufficiency in postpartum women.

Background: Lactation insufficiency is a prevalent challenge for nursing mothers globally. There is a growing interest in the use of herbal galactagogues for enhancing lactation, but their therapeutic efficacy and underlying mechanisms need thorough investigation. This study aims to investigate the efficacy and mechanisms of action of herbal galactagogues in addressing lactation insufficiency by utilizing real-world data and employing a network analysis approach. Methods: Our retrospective study used Taiwan's Longitudinal Health Insurance Database 2000 (LHID2000) to identify 490 patients diagnosed with lactation insufficiency from 2000 to 2018. We analyzed demographic characteristics, co-existing diseases, and prescription patterns for both users and non-users of Chinese herbal products (CHP). Additionally, we utilized a network analysis approach to explore potential compounds and targets in the most frequently used CHP, the Wang Bu Liu Xing and Lu Lu Tong herb pair (WLHP) combination. Results: Out of 490 patients, 81% were CHP users. There were no significant differences in demographic characteristics between CHP users and non-users, but we observed a notable divergence in the prevalence of co-existing diseases. A detailed examination of CHP prescriptions revealed the predominance of WLHP, prompting further investigation. Comprehensive analysis identified 29 major compounds in WLHP, which were associated with 215 unique targets. Intersection analysis revealed 101 overlapping targets between WLHP and lactation, suggesting their potential as therapeutic targets for lactation insufficiency treatment. Topological analysis of the protein-protein interaction (PPI) network identified 13 hub genes potentially crucial for the therapeutic effect of WLHP. Functional enrichment analysis showed that these targets were involved in critical lactation regulation pathways, including the PI3K-Akt signaling pathway, prolactin signaling pathway, estrogen signaling pathway, and AMPK signaling pathway. Discussion: This study emphasizes the potential of CHP, specifically the WLHP combination, in managing lactation insufficiency. The multi-compound, multi-target approach of WLHP and its interaction with key biological processes and signaling pathways offer valuable insights into the underlying mechanisms of its therapeutic effects. These findings warrant further experimental validation and can guide future research and clinical applications of CHP in lactation insufficiency treatment.

Integrated Chinese and Western Medicine for Coronary Heart Disease Prevention in Polycystic Ovary Syndrome: A 19-Year Nationwide Cohort Study.

Background: Polycystic ovarian syndrome (PCOS) is a common endocrine disorder in women of reproductive age. Chinese herbal medicine (CHM), a major branch of traditional Chinese medicine, has been reported to exert beneficial effects on PCOS symptoms. However, the relationship between CHM use and the risk of coronary heart disease (CHD) in patients with PCOS remains unclear. The study aimed to investigate the potential protective role of CHM on CHD incidence in patients with PCOS and to understand CHM prescription patterns for PCOS treatment. Methods: This retrospective cohort study was performed using data from Taiwan's Longitudinal Health Insurance Database 2000 (LHID2000). Patients with PCOS were identified, and data on their demographic characteristics, comorbidities, and medication use were extracted. The patients were then divided into CHM and non-CHM users. Results: Propensity score matching yielded two balanced cohorts of 2913 individuals each. CHM users demonstrated a 94% reduced risk of CHD compared to non-CHM users (adjusted hazard ratio, 0.06; 95% confidence interval, 0.04-0.1) during the 19-year follow-up period. Stratified analysis confirmed the benefits of CHM treatment on the incidence of CHD across various subgroups. The key elements identified in the CHM prescription patterns associated with reduced CHD risk were Xiang Fu, Yi Mu Cao, Jia Wei Xiao Yao San, and Gui Zhi Fu Ling Wan. Conclusion: This study provides evidence that CHM may protect against CHD development in patients with PCOS. It is also highlight the most frequently prescribed CHM for mitigating CHD risk among patients with PCOS. These findings lay the groundwork for future studies to confirm and elucidate CHM's therapeutic mechanisms in PCOS-related CHD prevention, offering a significant contribution to the field.

Acupuncture Is Effective at Reducing the Risk of Stroke in Patients with Migraines: A Real-World, Large-Scale Cohort Study with 19-Years of Follow-Up.

Migraines are common headache disorders and risk factors for subsequent strokes. Acupuncture has been widely used in the treatment of migraines; however, few studies have examined whether its use reduces the risk of strokes in migraineurs. This study explored the long-term effects of acupuncture treatment on stroke risk in migraineurs using national real-world data. We collected new migraine patients from the Taiwan National Health Insurance Research Database (NHIRD) from 1 January 2000 to 31 December 2017. Using 1:1 propensity-score matching, we assigned patients to either an acupuncture or non-acupuncture cohort and followed up until the end of 2018. The incidence of stroke in the two cohorts was compared using the Cox proportional hazards regression analysis. Each cohort was composed of 1354 newly diagnosed migraineurs with similar baseline characteristics. Compared with the non-acupuncture cohort, the acupuncture cohort had a significantly reduced risk of stroke (adjusted hazard ratio, 0.4; 95% confidence interval, 0.35-0.46). The Kaplan-Meier model showed a significantly lower cumulative incidence of stroke in migraine patients who received acupuncture during the 19-year follow-up (log-rank test, p < 0.001). Acupuncture confers protective benefits on migraineurs by reducing the risk of stroke. Our results provide new insights for clinicians and public health experts.

In Vivo Analysis of Extract of Leaves of Mistletoe as a Benalu Duku: Clinical Chemical Value Associated with Histopathology, Liver, Kidneys, and Lethal Dose Determinate.

The active compounds from the leaves of Dendrophthoe pentandra L. Miq., also known as, Benalu Duku (in Indonesia), are known to contain progesterone-like compounds (PLCs). This study aims to determine the effect of giving a single dose of PLCs on liver and kidney function in rats and the dose limit that causes the death of experimental animals. The PLCs were analyzed for chemical and physical characterization and compared to a pure standard of progesterone using HPLC, IR spectrometry, thermogravimetry, and NMR. The research was carried out in two sections. In section one, thirty-five healthy adult male rats were divided into six experimental groups and a control group of five rats each. The groups received, respectively, 50 to 75 mg/kg of PLCs (i.p.). The control group was given a 0.5 mL Aqua Pro injection. Alanine aminotransferase, aspartate aminotransferase, creatinine, and blood urea nitrogen were assessed using the clinical chemistry of blood serum analysis. Cell disruptions were analyzed to determine the degeneration effects of PLCs on the liver and kidney in the experimental and control groups. In section two, thirty healthy adult male rats were divided into 6 groups, each group of 5 rats, and injected with PLCs at a dose of 0.9-2.1 g/kg BW, followed by a lethal dose test. The control groups were available for 5 individual rats at 0 g/kg BW of PLCs. Our findings indicated that PLCs have a similarity chemical and physical characterized each other compounds, then the following administration of 50 to 75 mg/kg of PLCs did not affect the parameters of clinical chemistry. Histopathology analysis of the liver and kidney revealed normal subcellular levels in the experimental group, with the nonlethal dose at 0.9 g/kg BW.

Toxicity test of flavonoid compounds from the leaves of Dendrophthoe pentandra (L.) Miq. using in vitro culture cell models.

Background and Aim: The flavonoids from mistletoe are thought to have antimicrobial action. This encouraging finding supports the benefits of medicinal plants as a substitute for synthetic antimicrobials, thus promoting healthy lifestyles. In contrast, it is known that the use of topical drug formulations made from flavonoids of mistletoe (Dendrophthoe pentandra (L.) Miq. Loranthaceae) with Indonesian name, Benalu duku (BD) is required in skin cell irritation. This study aimed to assess the toxic effects of the flavonoid substances of BD, as an initial screening. Materials and Methods: A myeloma cell line was cultured in Roswell Park Memorial Institute medium, and the Baby Hamster Kidney clone 12 (BHK21) cell line was cultured in Dulbecco's Modified Eagle's Medium from stock (±9 × 107 cells/mL), and 1.2 mL of culture were distributed into each well of a microtiter plate. Subsequently, 0.2 mL of serially diluted flavonoid compounds (0.5-3 µg/mL) were added to 12 wells for each concentration, as trial groups (including control groups), followed by a 2-day incubation. Observations were performed based on the cytopathic effect (CPE) using an inverted microscope at a magnification of 100×. Results: Cytopathic effect was detected on the microtiter plate wells for the groups of myeloma and BHK21 cells at a flavonoid concentration of 0.5 µg/mL-3 µg/mL. Conclusion: Flavonoid compounds from BD were safely used for topical treatment of cancer cells at a concentration <2.491 µg/mL, whereas for non-cancerous cells, a concentration <2.582 µg/mL was sufficient (p < 0.05).

The Spatial Relationship and Surface Projection of Canine Sciatic Nerve and Sacrotuberous Ligament: A Perineal Hernia Repair Perspective.

Sciatic nerve entrapment can occur as post-operative complication of perineal hernia repair when sacrotuberous ligament is incorporated during hernia deficit closure. This results in sciatic sensory loss and paralysis of the hind leg. This study investigated the spatial relationship of sciatic nerve and sacrotuberous ligament and their surface topographic projection of 68 cadavers (29 Beagles and 39 Taiwanese mongrels) with various heights (25-56 cm). By gross dissection, the sacrotuberous ligament and sciatic nerve were exposed and their distance in between was measured along four parts (A, B, C, D) of sacrotuberous ligament. The present study revealed that the C was the section of sacrotuberous ligament where the sciatic nerve and the sacrotuberous ligament are closest to each other. Furthermore, a positive correlation was observed between C and height of the dogs. From the present study, we found that the C in smaller dogs has the shortest distance between the sciatic nerve and the sacrotuberous ligament, and thus the most vulnerable to sciatic nerve entrapment, and needs to be avoided or approached cautiously during perineal hernia repair.

The antimicrobial peptide pardaxin exerts potent anti-tumor activity against canine perianal gland adenoma.

Pardaxin is an antimicrobial peptide of 33 amino acids, originally isolated from marine fish. We previously demonstrated that pardaxin has anti-tumor activity against murine fibrosarcoma, both in vitro and in vivo. In this study, we examined the anti-tumor activity, toxicity profile, and maximally-tolerated dose of pardaxin treatment in dogs with different types of refractory tumor. Local injection of pardaxin resulted in a significant reduction of perianal gland adenoma growth between 28 and 38 days post-treatment. Surgical resection of canine histiocytomas revealed large areas of ulceration, suggesting that pardaxin acts like a lytic peptide. Pardaxin treatment was not associated with significant variations in blood biochemical parameters or secretion of immune-related proteins. Our findings indicate that pardaxin has strong therapeutic potential for treating perianal gland adenomas in dogs. These data justify the veterinary application of pardaxin, and also provide invaluable information for veterinary medicine and future human clinical trials.

US-like strain of porcine epidemic diarrhea virus outbreaks in Taiwan, 2013-2014.

Since late 2013, several outbreaks of porcine epidemic diarrhea virus (PEDV) infection have emerged in Taiwan. Suckling piglets under 2 weeks of age showed severe vomiting and watery yellowish diarrhea with morbidity and mortality ranging from 80 to 100% and 90 to 100%, respectively. A total of 68 samples from 25 pig farms were confirmed as positive for PEDV and negative for rotavirus and transmissible gastroenteritis virus by reverse transcription PCR, and the partial S gene of PEDV was analyzed. Phylogenetic analysis places all 18 Taiwanese PEDV isolates collected during this outbreak in the same clade as the US strains of PEDV. This novel PEDV is prevailing and currently causing severe outbreaks in Taiwan.

Neuronal Activity Stimulated by Liquid Substrates Injection at Zusanli (ST36) Acupoint: The Possible Mechanism of Aquapuncture.

Aquapuncture is a modified acupuncture technique and it is generally accepted that it has a greater therapeutic effect than acupuncture because of the combination of the acupoint stimulation and the pharmacological effect of the drugs. However, to date, the mechanisms underlying the effects of aquapuncture remain unclear. We hypothesized that both the change in the local spatial configuration and the substrate stimulation of aquapuncture would activate neuronal signaling. Thus, bee venom, normal saline, and vitamins B1 and B12 were injected into a Zusanli (ST36) acupoint as substrate of aquapuncture, whereas a dry needle was inserted into ST36 as a control. After aquapuncture, activated neurons expressing Fos protein were mainly observed in the dorsal horn of the spinal cord in lumbar segments L3-5, with the distribution nearly identical among all groups. However, the bee venom injection induced significantly more Fos-expressing neurons than the other substrates. Based on these data, we suggest that changes in the spatial configuration of the acupoint activate neuronal signaling and that bee venom may further strengthen this neuronal activity. In conclusion, the mechanisms for the effects of aquapuncture appear to be the spatial configuration changes occurring within the acupoint and the ability of injected substrates to stimulate neuronal activity.

Genetic characterization of type 2a canine parvoviruses from Taiwan reveals the emergence of an Ile324 mutation in VP2.

BACKGROUND: Canine parvovirus 2 (CPV 2) is a major infectious cause of mortality in puppies. The characteristic symptom of CPV 2 disease is intestinal hemorrhage with severe bloody diarrhea. Soon after CPV was first recognized in the late 1970s, the original virus, CPV 2, was replaced in the canine population by strains carrying minor antigenic variants (termed 2a, 2b, and 2c) of the VP2 gene that could be distinguished using monoclonal antibodies and molecular analyses. Here, we provide an updated molecular characterization of the CPV 2 circulating in Taiwan. METHODS: In this study, 28 isolates of CPV 2 from 144 dogs with suspected CPV infection were obtained from northern, central, and southern Taiwan from 2008 to 2012 and screened by PCR. The 28 isolates were sequenced, and a phylogenetic analysis of the VP2 gene was performed. RESULTS: Of the 28 Taiwanese CPV 2 isolates, 15 were identified as new CPV 2a, and 13 were identified as new CPV 2b. Compared to the reference CPV 2a, all 15 of the CPV 2a sequences collected in this study contain an Ile324 mutation caused by a TAT to ATT mutation at nucleotides 970-972 of the VP2 gene. CONCLUSION: Our VP2 sequence data revealed that both types are currently prevalent CPV 2 field strains circulating in Taiwan, and a unique Ile324 VP2 mutation was found in our Taiwanese CPV 2a isolates and recent Asian isolates. CPV 2c was not observed in this study.

The Possible Neuronal Mechanism of Acupuncture: Morphological Evidence of the Neuronal Connection between Groin A-Shi Point and Uterus.

Somatovisceral reflex suggested that the somatic stimulation could affect visceral function like acupuncture which treats diseases by stimulating acupoints. The neuronal connection between somatic point and visceral organ was not clear. Uterine pain referred to the groin region has long been recognized clinically. Wesselmann, using neurogenic plasma extravasation method, showed that uterine pain was referred to the groin region through a neuronal mechanism (Wesselmann and Lai 1997). This connection could be considered through the somatovisceral reflex pathway. However, the relay center of this pathway is still not clearly identified. In the present study, bee venom was injected in the groin region to induce central Fos expression to map the sensory innervation of groin region. Pseudorabies virus (PrV), a transneuronal tracer, was injected in the uterus to identify the higher motor control of the uterus. Immunohistochemistry staining revealed the Fos expression and PrV-infected double-labeled neurons in the nucleus of solitary tract (NTS), the dorsal motor nucleus of vagus (DMX), and the paraventricular hypothalamic nucleus (PVN). These results suggest a somatoparasympathetic neuronal connection (groin-spinal dorsal horn-NTS/DMX-uterus) and a somatosympathetic neuronal connection (groin-spinal dorsal horn-NTS-PVN-uterus). These two neuronal connections could be the prerequisites to the neuronal basis of the somatovisceral reflex and also the neuronal mechanism of acupuncture.

The spatial and segmental innervation of somatic acupoint - a study of canine Shen-Shu point (BL-23).

Although an acupuncture needle penetrates the skin, subcutaneous tissue, and underlying muscle, the most effective locus for the somatic acupoint on the needle path is not well established. We therefore investigated the sensory innervations of tissues in the needle path of the canine Shen-Shu point and evaluated their roles in initiating an acupunctural signal. Horseradish peroxidase solution was injected at all three levels within the acupoint. Only a few peroxidase-positive neurons were observed in the L1 dorsal root ganglion following intradermal injection. Following subcutaneous injection, peroxidase-labeled neurons were detected extending from spinal levels T10 to L2, with maximal labeling at T12 (46.3%). Approximately 95% of positive neurons were at spinal levels T11, T12, T13, and L1. As a result of an intramuscular injection, labeled neurons were observed at spinal levels T12 to L3, with most labeling occurring at L1 (39.9%). Approximately 95% of positive neurons were at spinal levels T13, L1, and L2. The results suggest that most afferent terminals are in the subcutaneous tissue rather than the muscular tissue, with an approximate ratio of 3.75:1. The data provide solid evidence that sensory innervation to a somatic acupoint is confined to a spinal segment and spatially organized, and we speculate that to cause a maximum effect, the centripetally transmitted signal from needling a somatic acupoint is spatio-segmental and divergently amplified.

The spinal cord connections of the myofascial trigger spots.

BACKGROUND: Recent electrophysiological studies revealed that endplate noise (EPN) could be specifically recorded from a myofascial trigger point (MTrP) region. EPN has been considered as the focal graded potentials due to excessive acetylcholine release in neuromuscular junction. A recent histological study has demonstrated a free nerve ending at the vicinity of the site, from where EPN could be recorded in an MTrP region. However, the sensory (afferent) and the motor (efferent) connections between an MTrP and the spinal cord still has never been fully studied. AIMS: The aim of this study was to delineate both motor and sensory connections between an MTrP and the spinal cord by applying the stain with horseradish peroxidase (HRP). METHODS: Nine Wistar rats were studied. When the rat was anesthetized, its biceps femoris muscles were exposed for localizing the myofascial trigger spot (MTrS, equivalent to MTrP in human). In one side, a monopolar Teflon-coated, hollow-needle electrode was used for searching EPN in an MTrS region, and then HRP was injected via this hollow-needle electrode into the site where EPN was recorded. HRP was also injected into a normal (non-taut band, non-MTrS) site in the contralateral side to obtain the control data. Two days after HRP injection, the rats were sacrificed and their spinal cords and dorsal root ganglia (DRG) were sectioned for the identification of the sites where neurons were labeled with HRP. RESULTS: The HRP-labeled neurons were found in the ventral horn of the spinal cord and in the DRG over L3, L4, and L5, while most were found in the L5 level. The mean numbers of HRP-labeled neurons in the EPN side looked smaller than that in the control side, but the difference did not reach statistically significant level (P>0.05). The mean values of the diameters of the HRP-labeled neurons in the DRG were not significantly different between the EPN side and the control side (P>0.05). However, HRP-neurons in the ventral horn of the spinal cord in the EPN side showed mild tendency to be smaller than that in the control side. CONCLUSIONS: The spinal cord connections of an MTrS are basically similar to that for a normal tissue region. The motor neurons related to MTrS tended to be smaller in their diameters. The findings in this study further supported the previously proposed hypotheses for the pathogenesis of an MTrP.

The regulatory mechanism of Tremella mesenterica on steroidogenesis in MA-10 mouse Leydig tumor cells.

Tremella mesenterica (TM), a yellow jelly mushroom, has been traditionally used as tonic food to improve body condition in Chinese society for a long time. We have previously demonstrated that TM reduced in vitro hCG-treated steroidogenesis in MA-10 mouse Leydig tumor cells without any toxicity effect. In the present study, the mechanism how TM suppressed hCG-treated steroidogenesis in MA-10 cells was investigated. MA-10 cells were treated with vehicle, human chorionic gonadotropin (hCG, 50 ng/ml), or different reagents with or without TM to clarify the effects. TM significantly suppressed progesterone production with the presences of forskolin (10 and 100 microM) or dbcAMP (0.5 and 1mM), respectively, in MA-10 cells (p<0.05), which indicated that TM suppressed steroidogenesis after PKA activation along the signal pathway. Beyond our expectation, TM induced the expression of steroidogenic acute regulatory (StAR) protein with or without hCG treatments. However, TM profoundly decreased P450 side chain cleavage (P450scc) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) enzyme activities without any influences on the expression of both enzymes. These inhibitions on steroidogenic enzyme activities might counteract the stimulation of StAR protein expression. In conclusion, results suggest that TM suppressed hCG-treated steroidogenesis in MA-10 cells by inhibiting PKA signal pathway and steroidogenic enzyme activities.

The in vivo effect of Cordyceps sinensis mycelium on plasma corticosterone level in male mouse.

Cordyceps sinensis (CS), an Ascomycetes fungus parasitic to Lepidoptera larvae, has been traditionally used as nutritious food for the enhancement in immuno-modulation in Chinese society for a long time. Previous report has demonstrated the CS water extract stimulates in vitro corticosterone production in rat primary adrenal cells. In the present studies, we determined the in vivo effects of CS and its fractions on plasma corticosterone production in mouse. Different concentrations of CS and CS fractions dissolved in water (0.02 and 0.2 mg/g body weight) were fed to immature and mature mice from 1, 3 or 7 d. The plasma levels of corticosterone were determined by radioimmunoassay (RIA), and the weight of adrenal gland and body weight were also evaluated. Results illustrated that plasma corticosterone levels were significantly induced by F2 at 0.02 mg/g body weight with 7 d feeding in immature mice, and by CS at 0.02 mg/g body weight with 3 d feeding and F3 at 0.02 mg/g body weight for 7 d feeding in mature mice, respectively (p < 0.05). There were no differences of adrenal gland weight except there was significant stimulation by CS at 0.2 mg/g body weight with 3 d feeding in mature mice (p < 0.05) and there were significant inhibitions by both dosages of F3 for 3 d feeding in immature mice and F2 for 7 d feeding in mature mice (p < 0.05), respectively. Concerning body weight, the stimulatory effects were observed with CS feeding at 0.2 mg/g body weight for 7 d and F3 feeding at 0.02 mg/g body weight for 3 and 7 d in mature mice. Whereas, the inhibitory effect were observed in F2 feeding at 0.2 mg/g body weight for 7 d in immature mice and at both dosages for 7 d in mature mice, respectively. Taken together, these studies illustrate that CS and its fractions stimulated mouse in vivo corticosterone production. However, CS and its fractions didn't have constant stimulatory or inhibitory effects on the weights of body and adrenal glands.

Effect of intrathecal octreotide on thermal hyperalgesia and evoked spinal c-Fos expression in rats with sciatic constriction injury.

This study was designed to determine whether intrathecal octreotide (sandostatin), a synthetic octapeptide derivative of somatostatin, relieved thermal hyperalgesia and reduced the evoked spinal c-Fos expression in rats with chronic constriction injury (CCI) of the sciatic nerve. Intrathecal catheters were implanted in rats 7 days before CCI of the sciatic nerve over the left hind limb. After confirmation of the development of thermal hyperalgesia by decreased paw withdrawal latencies (PWL) to heat stimulation 7 days after CCI, intrathecal sandostatin at 20, 40, and 80 microg was administered, respectively. Rats in the control group received saline injections intrathecally. PWLs were evaluated at 30, 60, 120, 180, and 240 min after drug administration. Detection of Fos-like immunoreactivity (Fos-LI) neurons in the dorsal horn of the spinal cord following drug administration was performed after mechanical stimulation (stroking of the hind paws) on the 14th day after CCI. The reduction of PWL was attenuated significantly in the groups that received intrathecal sandostatin at 20, 40, and 80 g when compared with the saline group. However, PWL did not return to pre-CCI values in all groups. In the 40 microg group, PWL returned up to 76% of pre-CCI values 120 min after drug administration. Stroking of the hind paw in CCI-treated (ipsilateral) limbs induced a significantly greater expression of spinal Fos-LI neurons than that of non-CCI treated (contralateral) limbs in each group. The number of Fos-LI neurons in animals receiving intrathecal sandostatin was dose-dependently reduced. Expression of Fos-LI neurons in the 80 microg group was nearly completely inhibited. These data suggest that intrathecal sandostatin significantly relieved thermal hyperalgesia behaviorally but with limited effects and dose-dependently reduced spinal Fos-LI neurons expression evoked by stroking stimulation, which may reflect mechanical allodynia in rats with sciatic constriction injury. This implies that intrathecal sandostatin was effective in the treatment of neuropathic pain.

Effect of botulinum toxin on endplate noise in myofascial trigger spots of rabbit skeletal muscle.

OBJECTIVE: To assess the effect of botulinum toxin type A (BTX-A) on the endplate noise prevalence in rabbit myofascial trigger spots to confirm the role of excessive acetylcholine release on the pathogenesis of myofascial trigger points and to develop an objective indicator of the effectiveness of BTX-A in the treatment of myofascial trigger points. DESIGN: Eighteen adult New Zealand rabbits were divided into three groups that received a single bolus of BTX-A over a myofascial trigger spot region on one side of the biceps femoris muscle. Another 10 rabbits received multiple-point injections in a myofascial trigger spot where endplate noises were found. A control study was performed on the other side of the biceps femoris muscle. The endplate noise prevalence in a myofascial trigger spot region was assessed. RESULTS: It was found that injection of BTX-A reduced the prevalence of endplate noise. No significant differences between a single bolus injection and multiple-point injections were noted, although there was some evidence that multiple-point injections might maintain the endplate noise decreasing effect much longer than a single injection. CONCLUSIONS: This study demonstrated the suppressive effect of BTX-A on endplate noise prevalence in a myofascial trigger spot region. The prevalence of endplate noise in the myofascial trigger point region may be a useful objective indicator for evaluating the therapeutic effectiveness of BTX-A injection to treat myofascial trigger points.