Integration of transcriptome and metabolome reveals the accumulation of related metabolites and gene regulation networks during quinoa seed development.

Quinoa seeds are gluten- and cholesterol-free, contain all amino acids required by the human body, have a high protein content, provide endocrine regulation, protein supplementation, and cardiovascular protection effects. However, metabolite accumulation and transcriptional regulatory networks in quinoa seed development are not well understood. Four key stages of seed development in Dianli-3260 and Dianli-557 were thus analyzed and 849 metabolites were identified, among which sugars, amino acids, and lipids were key for developmental processes, and their accumulation showed a gradual decrease. Transcriptome analysis identified 40,345 genes, of which 20,917 were differential between the M and F phases, including 8279 and 12,638 up- and down-regulated genes, respectively. Grain development processes were mainly enriched in galactose metabolism, pentose and glucuronate interconversions, the biosynthesis of amino acids, and carbon metabolism pathways, in which raffinose, phosphoenolpyruvate, series and other metabolites are significantly enriched, gene-LOC110689372, Gene-LOC110710556 and gene-LOC110714584 are significantly expressed, and these metabolites and genes play an important role in carbohydrate metabolism, lipid and Amino acid synthesis of quinoa. This study provides a theoretical basis to expand our understanding of the molecular and metabolic development of quinoa grains.

The Protective Effect of Bajijiasu on the Treatment of Hypertensive Nephropathy in Rats.

BACKGROUNDS: Hypertensive nephropathy (HN) is a kind of renal disease caused by essential hypertension that eventually worsens into end-stage renal disease (ESRD). HN could damage the renal tubules, induce kidney damage and renal failure, and increase the risk of stroke, heart disease or death, but there are few ideal drugs for HN treatment. METHODS: In this study, we explored the therapeutic effect of bajijiasu (a compound from Morinda officinalis how and a common traditional Chinese medicine for tonifying the kidney) on the HN rat model. Biochemical analysis, HE staining, and PAS staining were used to assess the effects of bajijiasu on HN rat model. Western blotting was used to analyze the potential mechanisms. RESULTS: The results of HE staining and PAS staining showed that bajijiasu could alleviate the pathological changes in HN rat models; biochemical analysis found that the concentration of Malondialdehyde (MDA), total protein (TP), albumin (ALB), microalbuminuria (MALB), blood urea nitrogen (BUN), creatinine (Cr), triglyceride (TG), and low-density lipoprotein-cholesterol (LDL-C) were significantly decreased compared with the model group after bajijiasu treatment; and bajijiasu could regulate the expression of TNF-α, IL-6, MDA, SOD1 and AGEs in HN rats; the result of western blotting demonstrated that bajijiasu could down-regulate the expression of TGFß1, NOX4, JNK, p- JNK and up-regulate the expression PPARγ and SOD 1 in HN rats. CONCLUSION: Those results demonstrated that bajijiasu could alleviate the pathological changes and physiological and biochemical symptoms of HN rat models by regulating the expression of TGFß1, PPARγ, JNK, p-JNK, NOX4 and SOD1 but could not lower the blood pressure of HN rats. Those pieces of evidence may provide a new therapeutic method for HN treatment.

Transcriptome and Metabolome Analyses Revealed the Response Mechanism of Quinoa Seedlings to Different Phosphorus Stresses.

Quinoa (Chenopodium quinoa Willd.) is a dicotyledonous annual herb of Family Amaranthaceae and Subfamily Chenopodiaceae. It has high nutritional and economic value. Phosphorus (P) is an essential plant macronutrient, a component of many biomolecules, and vital to growth, development, and metabolism. We analyzed the transcriptomes and metabolomes of Dianli-1299 and Dianli-71 quinoa seedlings, compared their phenotypes, and elucidated the mechanisms of their responses to the phosphorus treatments. Phenotypes significantly varied with phosphorus level. The plants responded to changes in available phosphorus by modulating metabolites and genes implicated in glycerophospholipid, glycerolipid and glycolysis, and glyconeogenesis metabolism. We detected 1057 metabolites, of which 149 were differentially expressed (DEMs) and common to the control (CK) vs. the low-phosphorus (LP) treatment samples, while two DEMs were common to CK vs. the high-phosphorus (HP) treatment samples. The Kyoto Encyclopedia of genes and genomes (KEGG) annotated 29,232 genes, of which 231 were differentially expressed (DEGs) and common to CK vs. LP, while one was common to CK vs. HP. A total of 15 DEMs and 11 DEGs might account for the observed differences in the responses of the quinoa seedlings to the various phosphorus levels. The foregoing results may provide a theoretical basis for improving the phosphorus utilization efficiency in quinoa.

Puerarin ameliorates allodynia and hyperalgesia in rats with peripheral nerve injury.

Puerarin is a major active ingredient of the traditional Chinese plant medicine, Radix Puerariae, and commonly used in the treatment of myocardial and cerebral ischemia. However, the effects of puerarin on neuropathic pain are still unclear. In this study, a neuropathic pain animal model was created by partial sciatic nerve ligation. Puerarin (30 or 60 mg/kg) was intraperitoneally injected once a day for 7 days. Mechanical allodynia and thermal hyperalgesia were examined at 1 day after model establishment. Mechanical threshold and paw withdrawal latency markedly increased in a dose-dependent manner in puerarin-treated rats, especially at 7 days after model establishment. At 7 days after model establishment, quantitative real-time reverse transcriptase-polymerase chain reaction results showed that puerarin administration reversed mRNA expression of transient receptor potential vanilloid 1 (Trpv1) and transient receptor potential ankyrin 1 (Trpa1) in a dose-dependent manner in dorsal root ganglion neurons after peripheral nerve injury. These results suggest that puerarin dose-dependently ameliorates neuropathic pain by suppressing Trpv1 and Trpa1 up-regulation in dorsal root ganglion of neuropathic pain rats.

The efficacy and safety of the fractional radiofrequency technique for the treatment of atrophic acne scar in Asians: A meta-analysis.

BACKGROUND: The fractional radiofrequency (fRF) technique is a recently emerged technique. However, the exact outcomes of fRF for treating atrophic acne scar in Asians are still unclear. OBJECTIVE: To compare the clinical outcomes of fRF with the fractional laser technique in Asians with atrophic acne scar. MATERIALS AND METHODS: The databases MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, and Chinese National Knowledge Infrastructure (CNKI) were searched. Main clinical outcomes were participant-reported scar improvement, investigator-reported scar improvement, post-inflammatory hyperpigmentation (PIH), erythema duration, scab duration, and the pain level. RESULT: Six randomized controlled trials were included in this meta-analysis. The scar improvement was similar in both groups regarding participant-reported scar improvement (p = 0.48) and investigator-reported scar improvement (p = 0.89). However, the incidence of PIH in fRF group was lower in comparison with the laser group (p < 0.001). The average duration of erythema was shorter in fRF group than in the laser group (p < 0.001). The mean time for debridement was shorter in fRF group than the carbon dioxide fractional laser system (p = 0.02). The pain level did not differ significantly in the two groups (p = 0.53). CONCLUSION: Although some bias exists in our study, fRF appears to be a superior alternative for the treatment of atrophic acne scar in Asians.

Comparison of Acupuncture Effect on Blood Perfusion between Needling Nonacupoint on Meridian and Needling Nonacupoint off Meridian.

To verify the ancient theory of rather missing the acupoint than missing the meridian, acupuncture at nonacupoint on meridian and acupuncture at nonacupoint off meridian were performed, respectively. The blood perfusion (BP) on the calf around bladder meridian area was measured with a laser Doppler perfusion imager before, during, and after acupuncture. The whole scanning field was divided into seven subareas, and mean BP on each area was calculated. The ratio of mean BP between a subarea and a reference subarea was gotten, and then the change rate was calculated as ratio change rate (RCR). The results showed that RCR on bladder meridian area and around Chengshan (BL57) during or after acupuncture at nonacupoint on meridian was significantly higher than that at nonacupoint off meridian, which supports the ancient theory. Such differences may be attributable to some factors that can facilitate the signals transmission and produce a better acupuncture effect, such as richer nerve terminals, blood vessels, and mast cells which can produce stronger signals on the acupoints and the low hydraulic resistance channel along meridians which plays a role of signal transmitting channel to get a better effect of acupuncture.

[Protective effect of Pinus massoniana bark extracts against cisplatin-induced nephrotoxicity in human embryonic kidney cells].

OBJECTIVE: To investigate the in vitro protective effect of Pinus massoniana bark extracts (PMBE) against cisplatin-induced nephrotoxicity in human embryonic kidney cells (HEK293), and preliminarily study its mechanism. METHOD: Human embryonic kidney cells (HEK293) were cultured in vitro. The MTT assay was adopted to test the effect of PMBE and cisplatin on growth of HEK293 cells, and the protective effect of PMBE on cisplatin-induced nephrotoxicity of HEK293, and then detect the intracellular reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH) content, catalase (CAT), superoxide dismutase (SOD) and activity of thioredoxin reductase (TrxR). RESULT: PMBE could promote growth of HEK293 cells at low concentrations, but generate slight nephrotoxicity at high concentration. Cisplatin could inhibit growth of HEK293 cells, increase ROS and MDA content, while reducing SOD, CAT and TrxR. The pre-protective PMBE was added to reduce cisplatin's injury to HEK293 cells, ROS, MDA and GSH content, SOD, CAT and TrxR within certain range. CONCLUSION: PMBE at specific concentration has the protective effect in cisplatin-induced nephrotoxicity in HEK293 cells. Its mechanism may be related to PMBE's antioxidant activity.

Expression of genes encoding cytokines and corticotropin releasing factor are altered by citalopram in the hypothalamus of post-stroke depression rats.

OBJECTIVES: To establish a rat model of post-stroke depression (PSD), and examine expression of genes encoding corticotropin releasing factor (CRF), interleukin 1 beta (IL-1ß), and tumor necrosis factor alpha (TNF-α) in the hypothalamus of PSD rats. METHODS: Rats were subjected to middle cerebral artery occlusion (MCAO) and chronic mild unpredictable stress (CUMS). Open field test and sucrose preference were used to examine depressive-like behaviors. Observed changes in gene expression levels in the hypothalamus of PSD rats were evaluated. RESULTS: MCAO with CUMS resulted in reduction of sucrose preference and locomotor activity. Genes encoding TNF-α, IL-1ß and CRF were highly expressed in the hypothalamus of rats subjected to MCAO and CUMS. The antidepressant citalopram, a selective serotonin reuptake inhibitor, had inhibitory effects on the expression of the aforementioned genes. We observed a correlation between CRF and IL-1ß mRNA levels in the citalopram-treated group of rats. CONCLUSION: The etiology of PSD is associated with cytokine expression in the hypothalamus and with hypothalamic-pituitary-adrenal axis activity. Citalopram administration inhibited the expression of TNF-α and IL-1ß transcripts in the hypothalamus, suggesting that selective serotonin reuptake inhibitors may be appropriate for PSD therapy.

Antitumor effects of Pinus massoniana bark extract in murine sarcoma S180 both in vitro and in vivo.

Pinus massoniana bark extract (PMBE) is a mixture of flavonoids, and showed a capability of inducing cell apoptosis; however, its properties have not yet been fully investigated. This paper evaluates the antitumor effects of PMBE in murine sarcoma S180 both in vitro and in vivo. In vitro, the growth inhibition of S180 cells was concentration dependent on PMBE as shown by the CCK-8 assay. The AO/EB staining and flow cytometry assay showed that PMBE induced S180 cell apoptosis. Cell cycle analysis revealed that the cells in the S phase were decreased by treatment with PMBE. In vivo, the treatment of 100, 200, and 300 mg/kg PMBE reduced the tumor weight and volume of S180-bearing NIH mice by 9%-67% and 13%-68%, respectively. Peripheral leukocyte count and lymphoproliferation were increased significantly after treatment with PMBE. Our results suggest that PMBE inhibits the tumor cell growth by inducing cell apoptosis and improving lymphoproliferation.

Pinus massoniana bark extract protects against oxidative damage in L-02 hepatic cells and mice.

The hepatoprotective activity of Pinus massoniana bark extract (PMBE) against hydrogen peroxide (H2O2)-induced damage in normal human liver L-02 cells and carbon tetrachloride (CCl4)-induced acute hepatotoxicity in mice was investigated. The L-02 cells were pre-treated with PMBE for 24 hours prior to exposure to 0.5 mM H2O2 for 3 or 24 hours. The cell viability, level of malondialdehyde (MDA) and glutathione (GSH), and the catalase (CAT) activity were evaluated. For in vivo experiments, mice were divided into groups and PMBE administered orally, after which each group was assigned a further treatment. Histopathological examination, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and GSH, the liver tissue levels of MDA and GSH, the activities of CAT and glutathione peroxidase (GSH-Px), were evaluated. PMBE treatment decreased the level of MDA and increased the cell viability, GSH content and CAT activity in H2O2 treated L-02 cells treated for 3 hours. PMBE obviously decreased serum ALT, AST, ALP, and liver tissue MDA, while increasing serum GSH, and liver tissue CAT and GSH-Px activities. In conclusion, PMBE treatment prevents H2O2 and CCl4-induced liver damage, and therefore could have a potential clinical usage.

Pinus massoniana bark extract selectively induces apoptosis in human hepatoma cells, possibly through caspase-dependent pathways.

Pinus massoniana bark extract (PMBE) is a mixture of flavonoids, whose antioxidant and apoptosis-inducing properties have been confirmed in vitro. In this study, the apoptotic effect and mechanism of PMBE in HepG2 human hepatoma cells were evaluated. PMBE exerted dose- and time-dependent cell growth inhibition on HepG2 cells, and selectively induced apoptosis without impact on normal liver L-02 cells. Apoptosis induced by PMBE in HepG2 cells was also confirmed by annexin-V/PI staining, transmission electron microscopy and sub-G1 phase accumulation. Moreover, PMBE also slightly blocked the cell cycle in the G2/M and S phases in HepG2 cells. The investigation of the mechanism by which PMBE induced apoptosis in HepG2 cells indicated that activation of extrinsic and intrinsic caspase, inhibition of NF-kappaB activation and decrease of the antiapoptotic protein Bcl-2 and the intact Bid protein were involved. Furthermore, the antitumor activity of PBME was demonstrated in vivo by a 42.88-69.94% reduction rate of tumor weight in H22 tumor-implanted mice. Taken together, these data indicate that PMBE selectively induces apoptosis in HepG2 cells through caspase-dependent pathways, and inhibits tumor growth in vivo, making it a potential candidate for anticancer therapeutics.

Effect of Pinus massoniana bark extract on IFN-gamma-induced ICAM-1 expression in HaCaT human keratinocytes.

AIMS OF THE STUDY: Pinus massoniana bark extract (PMBE) with known anti-oxidant activity is comprised of various flavonoids including several bioactive compounds. We found that PMBE contains 1.27% taxifolin, a well-studied compound with known anti-inflammatory activity. Therefore, in the present study, we evaluated the effects of PMBE and taxifolin on intercellular adhesion molecule-1 (ICAM-1) expression. MATERIALS AND METHODS: PMBE and taxifolin were prepared. After HaCaT cells were pre-treatmented with PMBE and taxifolin, HaCaT cells were treatmented with 1000 U/ml IFN-gamma for 24 h. RESULTS: Treatment of HaCaT cells with 1000 U/ml IFN-gamma for 24 h markedly increased ICAM-1 expression. However, PMBE pre-treatment (40 microg/ml for 24 h) significantly inhibited IFN-gamma-induced ICAM-1 expression. In equal concentrations of taxifolin, PMBE-mediated inhibition of ICAM-1 mRNA and protein expression was greater than taxifolin mediated-inhibition, and the front on inhibition of ICAM-1 protein expression was 2.24-2.30-fold of the latter. When cells were treated with both compounds at a concentration of 40 microg/ml, PMBE-mediated inhibition of ICAM-1 mRNA was also greater than taxifolin-mediated inhibition and PMBE on inhibition of ICAM-1 protein expression was 2.60-3.00-fold the inhibition mediated by taxifolin. CONCLUSIONS: PMBE including additional bioactive compounds may possibly synergize to inhibit transcription and translation of inducible ICAM-1expression and PMBE was greater than monomeric flavonoid taxifolin. These results indicate that PMBE exhibits great potential as a therapeutic treatment for inflammatory skin diseases.

Involvement of the Bcl-2 family members in Pinus massoniana bark extract induced apoptosis in HeLa cells.

Pinus massoniana bark extract (PMBE) contains a variety of flavonoids whose antioxidant properties have been confirmed in vitro. This study was undertaken to evaluate the cytotoxic effects and the mechanism of cell death on the PMBE-treated human cervical cancer cell line, HeLa. PMBE treatment led to cell growth inhibition in a dose- and time-dependent manner, and PMBE-induced apoptosis was confirmed by DAPI staining, TUNEL assays and sub-G1 phase accumulation. Cell cycle was also arrested in G2/M phase. Immunoblotting analysis showed that cytochrome c was released, the protein expression of Bax was increased, the protein expression of Bcl-2 was down-regulated and caspase-9 and -3 were activated in PMBE-treated HeLa cells. Taken together, PMBE inhibited proliferation, induces apoptosis and causes cell cycle arrest in HeLa cells, indicating that PMBE may be a potential therapeutic agent for cancer.

[Experimental verification of "treating lumbar-back problems by puncturing Weizhong (BL 40)" with blood perfusion imaging technique].

OBJECTIVE: To verify the classic saying of "Treating lumbar-back problems by puncturing Weizhong (BL 40)" in acu-moxibustion learning and to explore its underlying mechanism. METHODS: Forty volunteer sublects were asked to accept twice tests respectively, i.e., acupuncture of Weizhong (BL 40) and Yanglingquan (GB 34), for observing changes of skin blood perfusion (SBP) at the low back part of the body by using a laser Doppler perfusion imager. The whole observed area (lower back) was divided into five sub-areas, left and right areas involving Line-one and Line-two of the Bladder Meridian. RESULTS: In comparison with pre-acupuncture, SBP of both BL40 and GB34 groups increased significantly from 4 minutes' retention of needles on to 12 mm after withdrawing acupuncture needle (P < 0.01), but no significant difference was found between two groups in SBP (P > 0.05). The change rates of SBP at Line-one and Line-two of the Bladder Meridian on the same side of the stimulated Weizhong (BL 40) were significantly higher than those of Yanglingquan (GB 34) (P < 0.05), but no significant changes were found on the opposite side (both Line-one and Line-two) of the Bladder Meridian. CONCLUSION: Acupuncture of both BL4O and GB34 can increase SBP at the low back of the body (involving the Bladder Meridian) on the same side, but the changing rate of SBP of BL4O is hiaher than that of GB34.

[Re-discrimination on "reinforcement can be achieved by slow-yet-rapid needling, and rapid-yet-slow needling weakens evil qi"].

OBJECTIVE: To study on the original meanings of "reinforcement can be achieved by slow-yet-rapid needling, and rapid-yet-slow needling weakens evil-qi", one of important bases of reinforcing-reducing in acupuncture. METHODS: Using the principle of knowledge archaeology and through structure an d through structure analysis on the subjects described by "slow" and "rapid" and the papers appearing the words, the meanings of this sentence are re-discriminated. CONCLUSION: "Slow-rapid" prescribed in the reinforcing-reducing needling method in Huangdi's Internal Classic includes 3 subjects: pulse condition, needle-retained time, and velocity of withdrawing the needle. The "slow-rapid" in "reinforcement can be achieved by slow-yet-rapid needling, and rapid-yet-slow needling weakens evil-qi" should be pulse condition, but was not needling method, which means that the pulse changes from slow to rapid, marked by transformation of channels from deficiency to excess; the transform of the pulse from rapid to slow means that the channel transforms from excess to deficiency.

[Diuretic effect of injecting furosemide into low hydraulic resistance point Shuifen along the conception meridian in pigs with acute edema].

OBJECTIVE: To observe whether injection of medicine into low hydraulic resistance point along meridian brings about higher medicinal effect and to explore the efficacy of the theory that meridians are made up of channels featuring low hydraulic resistance by observing the diuretic effect of injecting furosemide or saline into the low hydraulic resistance point Shuifen (CV 9), vein and Zusanli (St 36) respectively. METHODS: Acute edema was induced in pigs by rapid intravenous injection of 2 000 ml normal saline. The pigs were divided into four groups: Shuifen (CV 9) injection of half dose furosemide group (SFF group), intravenous injection of full dose furosemide group (VF group), Zusanli (St 36) injection of full dose furosemide group (ZSLF group), and Shuifen (CV 9) injection of half dose normal saline group (SFS group). The accumulated urine quantity and the urine quantity generated in every 15-minute period were measured in each group respectively, every 15 minutes after injection, and the measurement lasted for two hours at one experiment. Each group involved eight times of experiments with one pig used for one experiment, which means the whole observation involved 32 times of experiments. RESULTS: The accumulated urine quantities observed in both SFF group and VF group were higher than those in the ZSLF group and the SFS group all through the measurement, showing significant differences during the period from the 15th minute to the 45th minute (P<0.05). But no significant difference was observed between the SFF group and the VF group during the whole 2-hour measurement (P>0.05). Analysis of urine quantity generated in every 15-minute period showed that diuretic effect climaxed during the 15th minute to the 30th minute in both SFF group and VF group. By contrast, ZSLF group reached diuresis climax during the 45th minute to 60th minute and no diuresis climax was observed in the SFS group all through the measurement. CONCLUSION: Injection of medicine into low hydraulic resistance point along meridian generates faster and more powerful medicinal potency, and this is likely to be applied to clinical practice. The theory that meridians are channels featuring low hydraulic resistance is important to the elucidation of meridians.

Effects of Pinus massoniana bark extract on cell proliferation and apoptosis of human hepatoma BEL-7402 cells.

AIM: To study the effects of Pinus massoniana bark extract (PMBE) on cell proliferation and apoptosis of human hepatoma BEL-7402 cells and to elucidate its molecular mechanism. METHODS: BEL-7402 cells were incubated with various concentrations (20-200 microg/mL) of PMBE for different periods of time. After 48 h, cell proliferation was determined by 3-(4,5-dimethyl-thiazolyl-2)-2,5-diphenyl tetrazolium bromide (MTT) assay. Apoptosis was evaluated by morphological observation, agarose gel electrophoresis, and flow cytometry analysis. Possible molecular mechanisms were primarily explored through immunohistochemical staining. RESULTS: PMBE (20-200 microg/mL) significantly suppressed BEL-7402 cell proliferation in a time- and dose-dependent manner. After treatment of BEL-7402 cells with 160 microg/mL PMBE for 24, 48, or 72 h, a typical apoptotic "DNA ladder" was observed using agarose gel electrophoresis. Nuclear condensation and boundary aggregation or split, apoptotic bodies were seen by fluorescence and electron microscopy. Sub-G1 curves were displayed by flow cytometry analysis. PMBE decreased the expression levels of Bcl-2 protein in a time-dependent manner after treatment of cells with 160 microg/mL PMBE. CONCLUSION: PMBE suppresses proliferation of BEL-7402 cells in a time- and dose-dependent manner and induces cell apoptosis by possibly downregulating the expression of the bcl-2 gene.

Potential biomedical properties of Pinus massoniana bark extract.

This study examined the potential biomedical properties of a bark extract from Pinus massoniana Lamb (PMBE). Its antioxidant activity, superoxide anion radical scavenging and free radical scavenging activities were examined. The antioxidant activity in a linoleic acid emulsion increased with increasing amounts of PMBE, with 100, 300 and 500 microg PMBE inhibiting 64.7%, 68.6% and 76.6% of peroxidation, respectively. Similarly, the radical scavenging activity by PMBE increased in a dose-dependent manner, suggesting that there may be a correlation between the antioxidant and scavenging activities. In addition, the effect of PMBE on human hepatocellular carcinoma BEL-7402 cells and normal liver L-02 cells was investigated in vitro. PMBE appeared to inhibit selectively the growth of BEL-7402 and slightly promoted the growth of L-02. Taken together, the results indicate that PMBE is a natural antioxidant that could potentially be used as a food supplement or as a candidate precursor substance for new anticancer therapeutics.

Effects of Huangqi (Hex) on inducing cell differentiation and cell death in K562 and HEL cells.

Huangqi (Astragalus membranaceus), a traditional Chinese medicine, has been used to ameliorate side effects of cancer chemotherapy in China. However, little is known about its molecular mechanisms. Here we show that induction of K562 or HEL cells with 1.5 mg/ml of Huangqi (Hex) (Components extracted from Huangqi) for 3-5 d results in the expression of beta-globin gene in both cell lines and leads to terminal differentiation. Moreover, the apoptosis in HEL cells can be induced by increasing concentration of Huangqi (Hex) to 4.5 mg/ml for 3-5 d. Upregulation of Apaf-1, caspase-3 and acetylcholinesterase (AChE) in HEL cells may play a crucial role in the process of apoptosis. The prospect of inducing expression of adult (beta) globin gene and apoptosis selectively in cancer cells is obviously attractive from a therapeutic point of view.