Lead poisoning in a 6-month-old infant: a case report.

Background: Lead is a toxic element of the environment that leads to major complications once it enters the blood stream, affecting multiple organs and systems of the body. Methods: We present a case of a 6-month-old female infant diagnosed with lead poisoning after presenting for routine child health care. The child's mother denied that the infant had a history of exposure to lead-containing substances. After a month of calcium supplementation, the patient's blood lead level remained elevated. We then tested the blood lead level of the mother and father. The results showed that the blood lead level of the mother was 77.0 µg/L and that of the father was 36.9 µg/L. The high blood lead level of the mother attracted our attention. We found that the mother had been using an external traditional Chinese medicine, Hu Wang Fen, which contains lead. After the mother's discontinuation of use of the traditional medicine, the child was treated with symptomatic treatment and chelation therapy. Subsequently, the patient's blood lead level decreased significantly. Results: Lead toxicity can be a life-threatening problem because of its potential for severe complications. In children, there is no safe blood lead level, and the toxic effects of lead can be prevented by the awareness and avoidance of traditional Chinese medicines that may contain lead. Conclusion: Even though the diagnosis of lead poisoning remains difficult in children, it must be taken into consideration by the clinician when treating a child using traditional Chinese medicines.

Parathyroid hormone (1-34) ameliorates cartilage degeneration and subchondral bone deterioration in collagenase-induced osteoarthritis model in mice.

AIMS: Parathyroid hormone (PTH) (1-34) exhibits potential in preventing degeneration in both cartilage and subchondral bone in osteoarthritis (OA) development. We assessed the effects of PTH (1-34) at different concentrations on bone and cartilage metabolism in a collagenase-induced mouse model of OA and examined whether PTH (1-34) affects the JAK2/STAT3 signalling pathway in this process. METHODS: Collagenase-induced OA was established in C57Bl/6 mice. Therapy with PTH (1-34) (10 µg/kg/day or 40 µg/kg/day) was initiated immediately after surgery and continued for six weeks. Cartilage pathology was evaluated by gross visual, histology, and immunohistochemical assessments. Cell apoptosis was analyzed by TUNEL staining. Microcomputed tomography (micro-CT) was used to evaluate the bone mass and the microarchitecture in subchondral bone. RESULTS: Enhanced matrix catabolism, increased apoptosis of chondrocytes in cartilage, and overexpressed JAK2/STAT3 and p-JAK2/p-STAT3 were observed in cartilage in this model. All of these changes were prevented by PTH (1-34) treatment, with no significant difference between the low-dose and high-dose groups. Micro-CT analysis indicated that bone mineral density (BMD), bone volume/trabecular volume (BV/TV), and trabecular thickness (Tb.Th) levels were significantly lower in the OA group than those in the Sham, PTH 10 µg, and PTH 40 µg groups, but these parameters were significantly higher in the PTH 40 µg group than in the PTH 10 µg group. CONCLUSION: Intermittent administration of PTH (1-34) exhibits protective effects on both cartilage and subchondral bone in a dose-dependent manner on the latter in a collagenase-induced OA mouse model, which may be involved in regulating the JAK2/STAT3 signalling pathway.Cite this article: Bone Joint Res 2020;9(10):675-688.

Flavonoid Myricetin Modulates GABA(A) Receptor Activity through Activation of Ca(2+) Channels and CaMK-II Pathway.

The flavonoid myricetin is found in several sedative herbs, for example, the St. John's Wort, but its influence on sedation and its possible mechanism of action are unknown. Using patch-clamp technique on a brain slice preparation, the present study found that myricetin promoted GABAergic activity in the neurons of hypothalamic paraventricular nucleus (PVN) by increasing the decay time and frequency of the inhibitory currents mediated by GABA(A) receptor. This effect of myricetin was not blocked by the GABA(A) receptor benzodiazepine- (BZ-) binding site antagonist flumazenil, but by KN-62, a specific inhibitor of the Ca(2+)/calmodulin-stimulated protein kinase II (CaMK-II). Patch clamp and live Ca(2+) imaging studies found that myricetin could increase Ca(2+) current and intracellular Ca(2+) concentration, respectively, via T- and L-type Ca(2+) channels in rat PVN neurons and hypothalamic primary culture neurons. Immunofluorescence staining showed increased phosphorylation of CaMK-II after myricetin incubation in primary culture of rat hypothalamic neurons, and the myricetin-induced CaMK-II phosphorylation was further confirmed by Western blotting in PC-12 cells. The present results suggest that myricetin enhances GABA(A) receptor activity via calcium channel/CaMK-II dependent mechanism, which is distinctively different from that of most existing BZ-binding site agonists of GABA(A) receptor.

Traditional Chinese medicine Bak Foong Pills alters uterine quiescence - possible role in alleviation of dysmenorrhoeal symptoms.

Since contractility of the uterus appears to be the major source of pain during dysmenorrhoea, alleviation of the contractions is believed to be a possible treatment strategy. Bak Foong Pills, a traditional Chinese formulation for use in gynaecological disorders, has long been thought as effective in the treatment of dysmenorrhoeal symptoms. The present study thus aims to investigate whether ethanol extract of Bak Foong Pills (BFP-Ex) or its constituent herbs may have direct effects on alleviating dysmenorrhoeal symptoms by altering uterine tone. This was investigated using isolated uterine preparations and intracellular messenger analysis of adenylate cyclase, via [(3)H]-adenine assay, and calcium, with fluorometry imaging, in myometrial cultures. BFP-Ex can stimulate uterine relaxation following oxytocin-induced contractions ex-vivo. Attempted inhibition of BFP-Ex's relaxatory response with a nitric oxide inhibitor and adenylate cyclase inhibitor, however, had no significant effect, suggesting that most of BFP-Ex's relaxatory response was not due to increases in NO or cAMP. Further studies on tetramethylpyrazine (TMP), a major active ingredient of BFP-Ex, indicated that TMP could modulate intracellular calcium levels in favour of uteri relaxation. The ability of Bak Foong Pills to alleviate menstrual pain may be due to direct regulation of uterine tone.

Bak Foong protects dopaminergic neurons against MPTP-induced neurotoxicity by its anti-apoptotic activity.

Bak Foong pill (BFP) is a well-known traditional Chinese medicine used for treatment of various gynaecological disorders. In addition, it exerts beneficial effects on other functional systems including the central nervous system. In the present study, we have investigated the possible neuroprotective action of BFP upon the nigrostriatal dopaminergic system by examining its effect on the expression patterns of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the 1-methyl-4-phenyl-1,2,3,6-tetrahyrdropyridine (MPTP)-induced Parkinson's disease (PD) mouse model. MPTP significantly decreased TH and DAT mRNA levels in the striatum and midbrain of both female and male C57BL/6 mice. However, with BFP pre-treatment mice showed a reduced neurotoxicity, with TH and DAT mRNA levels either not affected by MPTP or affected to a lesser extent in the midbrain and striatum when compared to vehicle treated animals. Possible anti-apoptotic activity of BFP was further studied in a dopamine-secreting neuroendocrine cell line, PC12. In this assay, MPTP elevated the expression of a pro-apoptotic gene, Bax, while this expression was reduced by BFP pre-treatment. Flow cytometry results also revealed that the effect of MPTP-induced apoptosis in PC12 cell lines was significantly reduced by BFP. The present results suggest that BFP is able to protect dopaminergic neurons from neurotoxin-induced neuronal injury with anti-apoptotic activity being one of the possible mechanisms.

Anti-apoptotic activity of Bak Foong Pills and its ingredients on 6-hydroxydopamine-induced neurotoxicity in PC12 cells.

Bak Foong Pills (BFP), a traditional Chinese medicine used for centuries for the enhancement of women's health, was shown to display neuro-protective activity in the 1-methyl-4-phenyl-1,2,4,6,-tetrahydro-pyridine (MPTP)-induced mouse model in a previous study. In order to elucidate its mechanism of action, we investigated the anti-apoptotic properties of Bak Foong Pills and its main ingredients, including Panax ginseng, Angelica sinensis, Glycyrrhiza uralensis, and Ligusticum chuanxiong, in the 6-hydroxydopamine (6-OHDA)-treated PC12 cell model. The addition of the neurotoxin could cause significant cell death and reduction of cell proliferation, as shown in the results determined by MTT assay, nitric oxide (NO) measurement and flow cytometric propidium iodine (PI) staining analysis, while pre-treatment of PC12 cell with either BFP or its main ingredients prevented the toxicity to some degree. In addition, the neurotoxin caused an elevated activation of caspase-3, the key enzyme for activation of the cellular apoptotic cascade, whereas BFP or its main ingredients inhibited the activation of caspase-3. These results strongly indicate that BFP and its main ingredients may provide a useful therapeutic strategy for the treatment of neurodegenerative diseases, such as Parkinson's disease.

Neuroprotective effects of Bak Foong Pill in 1-methyl-4-phenyl-1,2,3,6-tetrahyrdropyridine (MPTP)-induced Parkinson's disease model mice.

Neuroprotective effects of estrogen and estrogen-like chemicals on neurodegenerative diseases, especially Parkinson's disease, have been well established. In the present study, we compared the effects of Bak Foong Pill (BFP), a well-known gynaecological tonic in China, and 17beta-estradiol, on dopamine transporter (DAT) and tyrosine hydroxylase (TH) gene expression patterns in ovariectomized, 1-methyl-4-phenyl-1,2,3,6-tetrahyrdropyridine (MPTP)-induced Parkinson's disease (PD) model mice, using multiplex reverse transcription-polymerase chain reaction (RT-PCR). MPTP, a specific dopaminergic neurotoxin, significantly decreased DAT and TH mRNA levels in the striatum, midbrain and cerebellum, but not the cortex, of C57BL/6 mice. However, MPTP-challenge with BFP pretreatment demonstrated reduced neurotoxicity, with DAT and TH mRNA levels either not affected by MPTP or affected to a significantly lesser extent in the midbrain and striatum as compared to the MPTP treated controls. 17beta-estradiol treatment prevented MPTP-induced reduction of DAT expression in striatum and midbrain, but failed to alter TH expression. These results suggest that BFP is able to protect dopaminergic neurons against MPTP-induced neuronal damage in a mechanism that is different from the protective effect of estrogen.

Improvement of barrier function and stimulation of colonic epithelial anion secretion by Menoease Pills.

AIM: Menoease Pills (MP), a Chinese medicine-based new formula for postmenopausal women, has been shown to modulate the endocrine and immune systems. The present study investigated the effects of MP and one of its active ingredients, ligustrazine, on epithelial barrier and ion transport function in a human colonic cell line, T84. METHODS: Colonic transepithelial electrophysiological characteristics and colonic anion secretion were studied using the short circuit current (ISC) technique. RT-PCR was used to examine the expression of cytoplasmic proteins associated with the tight junctions, ZO-1 (zonula occludens-1) and ZO-2 (zonula occludens-2). RESULTS: Pretreatment of T84 cells with MP (15 microg/mL) for 72 h significantly increased basal potential difference, transepithelial resistance and basal ISC. RT-PCR results showed that the expressions of ZO-1 and ZO-2 were significantly increased after MP treatment, consistent with improved epithelial barrier function. Results of acute stimulation showed that apical addition of MP produced a concentration-dependent (10-5,000 microg/mL, EC50 = 293.9 microg/mL) increase in ISC. MP-induced ISC was inhibited by basolateral treatment with bumetanide (100 micromol/L), an inhibitor of the Na+-K+-2Cl- cotransporter, apical addition of Cl- channel blockers, diphenylamine-2, 2'-dicarboxylic acid (1 mmol/L) or glibenclamide (1 mmol/L), but not 4, 4'-diisothiocyanostilbene-2, 2'-disulfonic acid or epithelial Na+ channel blocker, amiloride. The effect of MP on ZO-1 and ZO-2 was mimicked by Ligustrazine and the ligustrazine-induced ISC was also blocked by basolateral application of bumetanide and apical addition of diphenylamine-2, 2'-dicarboxylic acid or glibenclamide, and reduced by a removal of extracellular Cl-. CONCLUSION: The results of the present study suggest that MP and lligustrazine may improve epithelial barrier function and exert a stimulatory effect on colonic anion secretion, indicating the potential use of MP and its active ingredients for improvement of GI tract host defense and alleviation of constipation often seen in the elderly.

Effects of Bak Foong Pills and Menoease Pills on white blood cell distribution in old age female rats.

This study examined the effects of Bak Foong Pills (BFP) and the new BFP-derived post-menopause formula, Menoease Pills (MBFP), on the distribution of peripheral white blood cells (WBC) between BFP/MBFP-treated and non-treated rats. Eighteen months old female SD rats were used to mimic post-menopausal and old age animal models. The percentage distribution of lymphocytes, monocytes and granulocytes were measured using flow cytometry with and without treatments of BFP or MBFP. Results showed that WBC distribution in old age rats were significantly different from that of adult rats, suggesting that as the animal aged, their WBC distributions were altered. Old age rats were observed to have much lower percentages of lymphocytes, but higher percentages of granulocytes when compared to the adult rats, indicating possible attenuated immunity. Following treatment with BFP or MBFP, WBC populations were found to be redistributed back into the ranges observed in adult animals. Furthermore, MBFP, was found to alter WBC distribution in a dose-dependent manner. When compared to estrogen (E(2)), a well documented regulator of immune function, results showed that MBFP was able to show significantly greater effects on WBC redistribution compared to E(2). However, in ovariectomised (ovx) old age rats, neither MBFP nor E(2) treated groups showed any changes in WBC redistribution. These results indicate that MBFP may share similarities to E(2). Indeed, the effect of MBFP and E(2) seems to require intact ovaries, which are believed to be necessary for the modulation of WBC distributions and immune functions. Overall, our findings suggest that BFP and MBFP may be able to regulate WBC population in old age female rats, and thus, indicate their potential role on improving the attenuated immunity evident in post-menopausal and elderly women.

Effect of Bak Foong pills on exocrine pancreatic-bile secretion.

We have recently demonstrated that Bak Foong Pills (BFP), a well-known Chinese medicine widely used for treating gynecological disorders, stimulates human colonic epithelial anion secretion, which was mediated by intracellular cAMP and Ca(2+). The present study further investigated the effect of BFP on exocrine pancreatic-bile secretion using in vivo and in vitro approaches. Duodenal infusion of BFP ethanol extract (1 g/kg) in rats produced increases in the volume and protein output of pancreatic-bile juice, but did not affect its pH. Surgical ablation of vagal neural pathway slightly reduced the effect of BFP on the protein output and volume, indicating that the vagal nerve pathway was not the major player in medicating the effect of BFP on exocrine pancreatic-bile secretion. Using CAPAN-1 cell line, a human pancreatic duct cell line, in conjunction with the short-circuit current (I(SC)) measurements, we further demonstrated that BFP could directly stimulate pancreatic HCO(3)(-) secretion. Basolateral addition of BFP (600 microg/ml) produced averaged charges transported of 2100+/-382.5 microC/cm(2), which was blocked by apical addition of Cl(-) channel blocker. Removal of HCO(3)(-) from the Krebs-Henseleit (K-H) solution inhibited the BFP-induced I(SC) by more than 95%. The present results suggest that BFP could improve digestive function by stimulating pancreatic protein and HCO(3)(-) secretion.

Cardiovascular protective effects of traditional Chinese medicine bak foong pills in spontaneously hypertensive rats.

The cardiovascular protective effects of the traditional Chinese medicine Bak Foong Pills (BFP) were investigated. Spontaneously hypertensive rats (SHR) were treated (3 g/kg) over a 5-month period and blood pressure measurements periodically tested with a plethysmographic tail cuff. Following treatment, blood samples were analysed for serum electrolyte levels and lipid levels and brain tissue subjected to micro-array analysis. In vitro experiments were also conducted to identify possible direct vasorelaxatory effect. The results showed that BFP was able to significantly reduce both systolic and diastolic blood pressure by about 30 mmHg in SHR following 5 months of treatment, when compared to untreated animals. Investigation for possible mechanisms of actions revealed that BFP treated rats had elevated blood serum K(+) levels, and also demonstrated decreased serum triglyceride levels. Micro-array analysis of brain tissue showed altered expression of acetylcholine and lysosphingolipid receptor genes that are known to regulate blood pressure. In vitro experiments also showed that BFP caused a concentration-dependant vasorelaxation of isolated rat aortae when contracted with phenylepherine, which was partially inhibited by nitric oxide synthase inhibitor L-NAME (100 microM). These data suggest that BFP is able to significantly reduce hypertension in SHR through mechanisms probably involving a combination of increased serum K(+), vasorelaxatory action, reduced serum triglyceride and altered gene regulation in the higher centres.

Effect of Bak Foong pills on enhancing dopamine release from the amygdala of female rats.

To investigate whether Bak Foong Pills (BFP), a well-known gynaecological tonic, has a direct effect on the central nervous system, we employed the in vivo electrochemical detection technique, fast cyclic voltammetry (FCV), to measure the dopamine release from the mesolimbic structure-amygdala of both male and female rats. The results showed that intracerebroventricular BFP (0.75, 1.5 microg) treatment promoted dopamine release from the amygdala in both female and ovariectomized female rats. The BFP-induced response appeared within 5 min after addition of BFP and lasted for at least 40 min. However, no effect of BFP was observed in male rats for an observed period of up to 60 min. The results suggest that BFP may have gender-specific beneficial effect on dopaminergic functions of the amygdala.

Effects of Bak Foong Pill on blood coagulation and platelet aggregation.

The effects of Bak Foong Pill (BFP, also known as Bai Feng Wan), a preparation of crude drugs in wide clinical use for treatment of gynecological disorders, on blood coagulation and platelet aggregation were investigated. The anticoagulant effect of BFP was evaluated by using thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (APTT) assays. Results showed that BFP 70% ethanol extract (BFP-E-ext) significantly prolonged the TT in a dose-dependent manner with values of 17.6, 38.3, and 50.4 s at concentrations of 4.0, 6.0, and 12.0 mg/ml, respectively. Whereas, the BFP-E-ext did not show significant prolonging effect in PT and APTT assays. The results suggest that the anticoagulant effect of BFP is mediated by directly blocking thrombin, the key enzyme in the blood coagulation cascade. BFP-E-ext significantly inhibited platelet aggregation induced by collagen and adenosine diphosphate (ADP) with inhibition percentages of 74 and 52% at a concentration of 6.0 mg/ml, respectively, whereas, it exhibited a weak inhibitory activity on platelet aggregation induced by archidonic acid (AA). Comparing to BFP-E-ext, the effects of BFP aqueous extract (BFP-W-ext) on both anticoagulant and antiplatelet activities were significantly less potent. Moreover, the effects of the 26 ingredients of BFP on blood coagulation and platelet aggregation were separately evaluated with 19 ingredient herbs exhibiting anticoagulant effect and 10 exhibiting antiplatelet effect. The anticoagulant and antiplatelet effects of BFP were collectively demonstrated by in vivo assays showing prolonged bleeding times after BFP treatment for two weeks. The results of the present studies may provide explanations for beneficial effects of BFP on the circulation and indicate its potential use for cardiovascular diseases.

Bak Foong Pills stimulate anion secretion across normal and cystic fibrosis pancreatic duct epithelia.

The present study examined the effect of Bak Foong Pills (BFP), an over-the-counter traditional Chinese medicine (China registration no. Z980035), on anion secretion and the underlying signaling pathways in normal and cystic fibrosis pancreatic duct cell lines, CAPAN-1 and CFPAC-1, respectively, using the short-circuit current technique. Apical addition of BFP ethanol extract (600 microg/ml) induced a fast transient I(SC) peak that was followed by a slower but more sustained increase in I(SC) in CAPAN-1 cells. However, the response to BFP in CFPAC-1 was predominantly the first transient peak. Apical addition of DIDS (200 microM) inhibited the first peak by more than 60% in both cell lines without significantly affecting the second I(SC) rise. More than 85% of the BFP-induced first transient in both cell lines was inhibited when extra and intracellular Ca(2+) was chelated or emptied by pre-treatment with BAPTA (100 microM) and thapsigargin (10 microM), respectively. Acute addition of PMA (1 microM), a PKC activator, blocked more than 95% of the BFP-induced first peak in both cell lines, consistent with previously reported PKC modulation of Ca(2+)-dependent pancreatic anion secretion. The BFP-induced second I(SC) rise in CAPAN-1 could be inhibited by 73.6% and 71.13% by pretreatment of the cells with MDL-12330A (20 microM), an adenylate cyclase inhibitor and Rp-cAMP (200 microM), a cyclic AMP antagonist, respectively. However, less than 25% of the I(SC) was inhibited by combined treatment with BAPTA and thapsigargin. The second rise was also completely blocked by DPC (2mM) or Glibenclamide (1mM). The results indicate that BFP ethanol extract stimulates pancreatic duct anion secretion in normal and CF cells via different signaling pathways involving both Ca(2+) and cAMP.