A dual organelle-targeting photosensitizer based on curcumin for enhanced photodynamic therapy.

The efficiency of photodynamic therapy (PDT) is related to the subcellular localization of photosensitizers (PSs) because organelles are associated with many fundamental life-sustaining activities. In this work, we synthesized a PS (CN) based on curcumin (CUR) and obtained enhanced PDT efficiency by simultaneously targeting lipid droplets (LDs) and the endoplasmic reticulum (ER). Compared with CUR, CN with a D-π-A-π-D structure possessed stronger intramolecular charge transfer features, resulting in longer absorption and emission wavelengths. In cell imaging experiments of CN using a confocal laser scanning microscope, a bright green emission in LDs and a weak orange emission in the ER were simultaneously observed due to its sensitivity to polarity. Surprisingly, CN with low singlet oxygen yields (0.13) exhibited an excellent photodynamic effect. Further experimental results showed that the phototoxicity of CN resulted in apoptosis by destroying the ER and ferroptosis by oxidizing polyunsaturated fatty acids (PUFAs) in LDs. This work paves the way for developing more effective photosensitizers with superior dual-targeting specificity.

Lipid droplet targeting-guided hypoxic photodynamic therapy with curcumin analogs.

Two photosensitizers (CCOH and CCN) were designed and synthesized by introducing coumarin into the curcumin (CUR) structure. Compared with CUR, more reactive oxygen species (ROS) were generated by CCOH and CCN in type I and II synergy upon light irradiation. Cell experiments indicated that CCN with an excellent LD-targeting effect could be used to monitor the changes in the morphology and number of LDs in tumor cells during PDT.

Treatment with 1,25-Dihydroxyvitamin D3 Delays Choroid Plexus Infiltration and BCSFB Injury in MRL/lpr Mice Coinciding with Activation of the PPARγ/NF-κB/TNF-α Pathway and Suppression of TGF-ß/Smad Signaling.

Neuropsychiatric systemic lupus erythematosus (NPSLE) is a serious complication of systemic lupus erythematosus (SLE) involving the nervous system with high morbidity and mortality. A key hypothesis in NPSLE is that a disrupted barrier allows autoantibodies and immune components of peripheral blood to penetrate into the central nervous system (CNS), resulting in inflammation and damage. The blood cerebrospinal fluid barrier (BCSFB), which consists of the choroid plexus and the hypothalamic tanycytes, has long been regarded as an immunological sanctuary site. 1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3] is the active form of vitamin D, which plays multiple roles in inflammation and immunoregulation. In this study, we investigated the possible protective effects of 1,25-dihydroxyvitamin D3 against BCSFB dysfunction in NPSLE in MRL/lpr mice and explored the mechanism by which 1,25-dihydroxyvitamin D3 inhibits the progression of NPSLE. In this study, we found that supplementation with 1,25-dihydroxyvitamin D3 markedly improved serological and immunological indices, delayed inflammatory infiltration, delayed neuronal deformation, and upregulated the expression of brain-derived neurotrophic factor (BDNF) proteins in the brain. Furthermore, 1,25-dihydroxyvitamin D3 downregulated proinflammatory cytokines such as nuclear factor kappa-B (NF-κB) and tumor necrosis factor-α (TNF-α) by activating peroxisome proliferator-activated receptor γ (PPARγ), and it reduced the expression of the TGF-ß/Smad signaling pathway. Our findings demonstrate that 1,25-dihydroxyvitamin D3 delayed cell infiltration into the choroid plexus and decreased markers suggestive of cognitive decline in MRL/lpr mice, and the mechanism may be related to protection against BCSFB disruption through activation of the anti-inflammatory PPARγ/NF-κB/TNF-α pathway as well as upregulation of BDNF and inhibition of the TGF-ß/Smad signaling pathway. These findings provide a novel direction for the study of NPSLE.

Struvite crystallization by using active serpentine: An innovative application for the economical and efficient recovery of phosphorus from black water.

Recovery of phosphorus from wastewater through struvite crystallization is one of the most attractive methods. However, the cost of chemical consumption makes this technology is unattractive to some extent. In this work, highly active serpentine was prepared by one-step mechanical activation and then used to recover phosphate as struvite from the black water containing 132.8 mg/L phosphorus and 3144 mg/L ammonia nitrogen. The results indicated that the prepared active serpentine can release magnesium ions and hydroxide ions simultaneously into an aqueous solution and is an ideal raw material for struvite crystallization. The factors for phosphorus recovery in this process mainly include mechanical activation intensity, serpentine dosage, and contact time. For the actual black water, a high recovery rate of phosphorus (>98%) is achieved by using active serpentine as the magnesium and alkali source for struvite precipitation. The recovery product was identified as struvite with a median particle size of 32.96 µm. It was confirmed that the mechanical activation damaged the crystal structure of the raw serpentine, improving the activity of Mg2+ and OH-. The undissolved Si-containing particles act as crystal seeds, accelerating the struvite crystallization process. Furthermore, a pilot-scale test was conducted with a rural public toilet in Xiong'an New District, Hebei Province. The results showed that an acceptable phosphorus recovery (98%) could be achieved using active serpentine. Additionally, it was demonstrated that the serpentine process to recover phosphate as struvite reduced the cost by 54.4% in compared with an ordinary chemical process. The active serpentine is a promising dual source of magnesium and alkali for the phosphorus recovery by the struvite method. It has a potential prospect for the large-scale application in phosphorus recovery and struvite fertilizer production.

Nitrogen and phosphorous recycling from human urine by household electrochemical fixed bed in sparsely populated regions.

Decentralized treatment of human urine in sparsely populated regions could avoid the problem of sewage collection in traditionally centralized treatment schemes and simultaneously utilize the recovered N/P fertilizer in-situ to nurture gardens. Herein, an integrated electrochemical fixed bed packed with divided magnesite and carbon zones was constructed for the pretreatment of human urine, followed by the recovery of 95.0% NH4+ and 85.8% PO43- via struvite precipitation and NH3 volatilization as well as the on-site employment of the produced struvite as fertilizer. In the process, the acid/base zones created by electrochemical water splitting dissolved the magnesite filler as the Mg2+ source of struvite, further creating an ideal pH environment for struvite precipitation and NH3 volatilization in the effluent. Without the need to control solution pH by chemical addition, the system can resist impacts from changes in water quality by adjustment of the current density and flow rate, indicating its great potential for automatic operation. Life cycle assessment indicated that the on-site employment of produced struvite avoids the long-distance fertilizer transportation required for fertilization, thus reducing carbon emission by a hundred million tons per year if the household facility is driven by clean electricity.

Genistein Alleviates High-Fat Diet-Induced Obesity by Inhibiting the Process of Gluconeogenesis in Mice.

Genistein is an isoflavone phytoestrogen that has been shown to improve obesity; however, the underlying molecular mechanisms involved therein have not been clearly elucidated. In this study, we administered genistein to high-fat diet-induced obese mice to investigate its effect on hepatic gluconeogenesis. The results showed that genistein treatment significantly inhibited body weight gain, hyperglycemia, and adipose and hepatic lipid deposition in high-fat diet-induced obese mice. Glucose tolerance test (GTT), insulin tolerance test (ITT) and pyruvate tolerance test (PTT) showed that genistein treatment significantly inhibited gluconeogenesis and improved insulin resistance in obese mice. In addition, this study also found that genistein could promote the expression of miR-451 in vitro and in vivo, and the dual-luciferase reporter system showed that G6pc (glucose-6-phosphatase) may be a target gene of miR-451. Both genistein treatment and in vivo injection of miR-451 agomir significantly inhibited gluconeogenesis and inhibited the expression of G6pc and Gk (glycerol kinase, a known target gene of miR-451). In conclusion, genistein may inhibit gluconeogenesis in obese mice by regulating the expression of Gk and G6pc through miR-451. These results may provide insights into the functions of miR-451 and food-derived phytoestrogens in ameliorating and preventing gluconeogenesis-related diseases.

Dietary betaine prevents obesity through gut microbiota-drived microRNA-378a family.

Betaine is a natural compound present in commonly consumed foods and may have a potential role in the regulation of glucose and lipids metabolism. However, the underlying molecular mechanism of its action remains largely unknown. Here, we show that supplementation with betaine contributes to improved high-fat diet (HFD)-induced gut microbiota dysbiosis and increases anti-obesity strains such as Akkermansia muciniphila, Lactobacillus, and Bifidobacterium. In mice lacking gut microbiota, the functional role of betaine in preventing HFD-induced obesity, metabolic syndrome, and inactivation of brown adipose tissues are significantly reduced. Akkermansia muciniphila is an important regulator of betaine in improving microbiome ecology and increasing strains that produce short-chain fatty acids (SCFAs). Increasing two main members of SCFAs including acetate and butyrate can significantly regulate the levels of DNA methylation at host miR-378a promoter, thus preventing the development of obesity and glucose intolerance. However, these beneficial effects are partially abolished by Yin yang (YY1), a common target gene of the miR-378a family. Taken together, our findings demonstrate that betaine can improve obesity and associated MS via the gut microbiota-derived miR-378a/YY1 regulatory axis, and reveal a novel mechanism by which gut microbiota improve host health.

Recovery phosphate and ammonium from aqueous solution by the process of electrochemically decomposing dolomite.

The cost-effective recovery of phosphate is of great significance to the mitigation of phosphorus resource depletion crisis. The electrochemical-decomposition of dolomite was developed to recover phosphate and ammonium from aqueous solution. The dolomite ore is mainly composed of CaMg(CO3)2 (53.73%), CaCO3 (28.93%) and SiO2 (16.59%). The continuous release of Mg2+ and Ca2+ were achieved by electrochemically decomposing dolomite ore, accompanied by the generation of base solution (9.0-10.5). The main factors affecting the recovery performance of phosphate (PO4-P) and ammonium (NH4-N) are current, initial concentration of PO4-P and NH4-N, initial pH of feed solution and feed rate. For a 30-d operation, the recovery rate of PO4-P was maintained at 90-97% and that of NH4-N at 50-60% under optimized operating conditions. The recovered product had low water solubility but high citric-acid-soluble, and was proposed as a slow-release fertilizer for crops. The proposed process as a simple, effective and green route may serve as a new strategy for recovering PO4-P and NH4-N from wastewaters.

Bidirectional regulation of genistein on the proliferation and differentiation of C2C12 myoblasts.

Genistein is a widely studied phytoestrogen. The effects of genistein on myoblasts were reported long ago, but the conclusions are controversial. In this study, we evaluated the effects of different concentrations of genistein on C2C12 myoblasts. Genistein treatment promoted myoblast proliferation in a dose-dependent manner in the concentration range of 0-2 µM/L, reaching its maximum effect at 2 µM/L. Proliferation then declined, and a concentration higher than 20 µM/L showed significant inhibition. In addition, genistein treatment promoted myoblast differentiation at a dose of 10 µM/L. However, at treatment concentrations higher than 10 µM/L, the effect on myoblast differentiation was rapidly inhibited as the concentration increased. Genistein treatment also down-regulated the expression of miR-222, resulting in increased expression of its target genes, MyoG, MyoD, and ERα and thereby promoting myoblast differentiation. Our results suggest that genistein has a dose-dependent and bidirectional regulation effect on myoblast proliferation and differentiation. We also found that genistein is a miRNA inducer, and it specifically affects the expression of miR-222 to regulate myoblast differentiation.

Guanidinoacetic Acid Regulates Myogenic Differentiation and Muscle Growth Through miR-133a-3p and miR-1a-3p Co-mediated Akt/mTOR/S6K Signaling Pathway.

Guanidinoacetic acid (GAA), an amino acid derivative that is endogenous to animal tissues including muscle and nerve, has been reported to enhance muscular performance. MicroRNA (miRNA) is a post-transcriptional regulator that plays a key role in nutrient-mediated myogenesis. However, the effects of GAA on myogenic differentiation and skeletal muscle growth, and the potential regulatory mechanisms of miRNA in these processes have not been elucidated. In this study, we investigated the effects of GAA on proliferation, differentiation, and growth in C2C12 cells and mice. The results showed that GAA markedly inhibited the proliferation of myoblasts, along with the down-regulation of cyclin D1 (CCND1) and cyclin dependent kinase 4 (CDK4) mRNA expression, and the upregulation of cyclin dependent kinase inhibitor 1A (P21) mRNA expression. We also demonstrated that GAA treatment stimulated myogenic differentiation 1 (MyoD) and myogenin (MyoG) mRNA expression, resulting in an increase in the myotube fusion rate. Meanwhile, GAA supplementation promoted myotube growth through increase in total myosin heavy chain (MyHC) protein level, myotubes thickness and gastrocnemius muscle cross-sectional area. Furthermore, small RNA sequencing revealed that a total of eight miRNAs, including miR-133a-3p and miR-1a-3p cluster, showed differential expression after GAA supplementation. To further study the function of miR-133a-3p and miR-1a-3p in GAA-induced skeletal muscle growth, we transfected miR-133a-3p and miR-1a-3p mimics into myotube, which also induced muscle growth. Through bioinformatics and a dual-luciferase reporter system, the target genes of miR-133a-3p and miR-1a-3p were determined. These two miRNAs were shown to modulate the Akt/mTOR/S6K signaling pathway by restraining target gene expression. Taken together, these findings suggest that GAA supplementation can promote myoblast differentiation and skeletal muscle growth through miR-133a-3p- and miR-1a-3p-induced activation of the AKT/mTOR/S6K signaling pathway.

The regulation of skeletal muscle fiber-type composition by betaine is associated with NFATc1/MyoD.

Increasing evidence indicates that muscular dysfunction or alterations in skeletal muscle fiber-type composition not only are involved in muscle metabolism and function but also can limit functional capacity. Therefore, understanding the mechanisms regulating key events during skeletal myogenesis is necessary. Betaine is a naturally occurring component of commonly eaten foods. Here, we showed that 10 mM betaine supplementation in vitro significantly repressed myoblast proliferation and enhanced myoblast differentiation. This effect can be mediated by regulation of miR-29b-3p. Further analysis showed that betaine supplementation in vitro regulated skeletal muscle fiber-type composition through the induction of NFATc1 and the negative regulation of MyoD expression. Furthermore, mice fed with 10 mM betaine in water for 133 days showed no impairment in overall health. Consistently, betaine supplementation increased muscle mass, promoted muscle formation, and modulated the ratio of fiber types in skeletal muscle in vivo. These findings shed light on the diverse biological functions of betaine and indicate that betaine supplementation may lead to new therapies for diseases such as muscular dystrophy or other diseases related to muscle dysfunction. KEY MESSAGES: Betaine supplementation inhibits proliferation and promotes differentiation of C2C12 myoblasts. Betaine supplementation regulates fast to slow muscle fiber-type conversion and is associated with NFATc1/MyoD. Betaine supplementation enhances skeletal myogenesis in vivo. Betaine supplementation does not impair health of mice.

Betaine Supplementation Enhances Lipid Metabolism and Improves Insulin Resistance in Mice Fed a High-Fat Diet.

Obesity is a major driver of metabolic diseases such as nonalcoholic fatty liver disease, certain cancers, and insulin resistance. However, there are no effective drugs to treat obesity. Betaine is a nontoxic, chemically stable and naturally occurring molecule. This study shows that dietary betaine supplementation significantly inhibits the white fat production in a high-fat diet (HFD)-induced obese mice. This might be due to betaine preventing the formation of new white fat (WAT), and guiding the original WAT to burn through stimulated mitochondrial biogenesis and promoting browning of WAT. Furthermore, dietary betaine supplementation decreases intramyocellular lipid accumulation in HFD-induced obese mice. Further analysis shows that betaine supplementation reduced intramyocellular lipid accumulation might be associated with increasing polyunsaturated fatty acids (PUFA), fatty acid oxidation, and the inhibition of fatty acid synthesis in muscle. Notably, by performing insulin-tolerance tests (ITTs) and glucose-tolerance tests (GTTs), dietary betaine supplementation could be observed for improvement of obesity and non-obesity induced insulin resistance. Together, these findings could suggest that inhibiting WAT production, intramyocellular lipid accumulation and inflammation, betaine supplementation limits HFD-induced obesity and improves insulin resistance.

Gypenosides Ameliorate Carbon Tetrachloride-Induced Liver Fibrosis by Inhibiting the Differentiation of Hepatic Progenitor Cells into Myofibroblasts.

Gypenosides (GPs), the predominant components of Gynostemma pentaphyllum, exert antifibrotic effects; however, the mechanisms underlying their ability to ameliorate liver fibrosis are unclear. Liver fibrosis was induced in C57BL/6 mice via subcutaneous injection of 10% carbon tetrachloride (CCl[Formula: see text] three times a week for two weeks. Then, CCl4 was administered in conjunction with intragastric GPs for another three weeks. For in vitro analyses, WB-F344, hepatatic progenitor cells (HPCs) were treated with transforming growth factor beta 1 (TGF-[Formula: see text]1) with or without GPs for 48[Formula: see text]h. The results showed that alanine aminotransferase (ALT) and aspartate transaminase (AST) activity, deposition of collagen, hydroxyproline content, and expression of alpha-smooth muscle actin ([Formula: see text]-SMA) and collagen type I (Col I) were significantly decreased after treatment with GPs ([Formula: see text], [Formula: see text], [Formula: see text], [Formula: see text]). In the 5M CCl4 group, the expression of HPC markers, Sox9 and cytokeratin 19 (CK19), was significantly increased compared with the normal or GPs-treated group ([Formula: see text], [Formula: see text]). Immunostaining showed that the number of Sox9 and [Formula: see text]-SMA double-positive cells was higher in the 5M CCl4 group than in the normal group, but the addition of GPs caused this cell number to decrease. In WB-F344 cells, the expression of [Formula: see text]-SMA and Col I was significantly increased after treatment with TGF-[Formula: see text], whereas in the GPs treatment group, expression was markedly decreased ([Formula: see text]). The levels of TGF-[Formula: see text] and TGF-[Formula: see text]R1 were markedly reduced after GPs treatment both in vivo and in vitro. In conclusion, GPs ameliorated CCl4-induced liver fibrosis via the inhibition of TGF-[Formula: see text] signaling, consequently inhibiting the differentiation of HPCs into myofibroblasts.

A sub-milligram-synthesis protocol for in vitro screening of HDAC11 inhibitors.

This work demonstrated the high efficiency of a sub-milligram-synthesis based medicinal chemistry method. Totally 72 compounds, consisting a tri-substituted pyrrolidine core, were prepared. Around 0.1mg of each compound was solid-phase synthesized. Based on the additive property of UV absorptions of unconjugated chromophores of a molecule, these compounds were quantified by UV measurement. A hit, whose IC50 value was 1.2µM in HDAC11 inhibition assays, highlights the applicability of the approach reported here in future optimization works.

Adaptability comparison of E. fetida in vermicomposting against sludge from livestock wastewater treatment plant based on their several growth stages.

Vermicomposting is a low-cost, eco-efficient process to deal with organic wastes. Mixtures of swine manure (SM), cow dung (CD), and animal wastewater treatment plant sludge (S) were applied as feeds, and Eisenia fetida was employed in this study to investigate the vermicomposting efficiency based on their several growth stages. The hatching test resulted in a 100 % hatching rate in S60SM40 (60 % S + 40 % SM) mixture, while 4.40 hatchlings per cocoon were observed. The growth of infancy performed best in 0-20 % CD mixtures (0.05 ± 0.002 g), followed by in SM + CD (0.04 ± 0.003 g). The highest growth rate of young and adult E. fetida was noticed in CD + S mixtures (11.14 ± 0.01 and 6.00 ± 0.02 mg/d/worm, respectively), while the higher cocoon production of adults was noticed in S + SM mixtures especially in S40SM60 (537 ± 5 worms). Moreover, the conversion of solids; the modified pH value; the reduction in total organic carbon (TOC); total Kjeldahl nitrogen (TKN), NH4-N, NO3-N, and C:N ratio; and the rich in total available phosphorus (TAP) and total potassium (TK) content by young and adult E. fetida were related to the growth of worms. Such work would benefit understanding and to increase the efficiency of vermicompost processing of different wastes.

Vermicomposting of sludge from animal wastewater treatment plant mixed with cow dung or swine manure using Eisenia fetida.

Vermicomposting of animal wastewater treatment plant sludge (S) mixed with cow dung (CD) or swine manure (SM) employing Eisenia fetida was tested. The numbers, weights, clitellum development, and cocoon production were monitored for 60 days at a detecting interval of 15 days. The results indicated that 100 % of the sludge can be the suitable food for growth and fecundity of E. fetida, while addition of CD or SM in sludge significantly (P < 0.05) increased the worm biomass and reproduction. The sludge amended with 40 % SM can be a great medium for the growth of E. fetida, and the sludge amended with 40 % CD can be a suitable medium for the fecundity of E. fetida. The addition of CD in sludge provided a better environment for the fecundity of earthworm than SM did. Moreover, vermicomposts obtained in the study had lower pH value, lower total organic carbon (TOC), lower NH4 (+)-N, lower C/N ratio, higher total available phosphorous (TAP) contents, optimal stability, and maturity. NH4 (+)-N, pH and TAP of the initial mixtures explained high earthworm growth. The results provided the theory basic both for management of animal wastes and the production of earthworm proteins using E. fetida.

Chronic acarbose treatment alleviates age-related behavioral and biochemical changes in SAMP8 mice.

The administration of maintaining the homeostasis of insulin/insulin-like growth factor 1 (IGF-1) signaling and/or glucose metabolism may reverse brain aging. In the present study, we investigated the effect of acarbose, an inhibitor of α-glucosidase, on age-related behavioral and biochemical changes. The SAMP8 mice were randomly divided into old control group and acarbose-treatment group. The mice in the acarbose group were administered acarbose (20 mg/kg/d, dissolved in drinking water) orally from 3 to 9 months of age when a new group of 3-month-old mice was added as young controls. The results showed that the aged controls exhibited declines in sensorimotor ability, open field anxiety, spatial and non-spatial memory abilities, decreased serum insulin levels, increased IGF-1 receptor and synaptotagmin 1 (Syt1) levels and decreased insulin receptor, brain-derived neurotrophic factor (BDNF) and syntaxin 1 (Stx1) levels in the hippocampal layers. The age-related behavioral deficits correlated with the serological and histochemical data. Chronic acarbose treatment relieved these age-related changes, especially with respect to learning and memory abilities. This protective effect of acarbose on age-related behavioral impairments might be related to changes in the insulin system and the levels of BDNF, IGF-1R, and the pre-synaptic proteins Syt1 and Stx1. In conclusion, long-term treatment with acarbose ameliorated the behavioral deficits and biochemical changes in old SAMP8 mice and promoted successful aging. This study provides insight into the potential of acarbose for the treatment of brain aging.

Lactation-related microRNA expression profiles of porcine breast milk exosomes.

Breast milk is the primary source of nutrition for newborns, and is rich in immunological components. MicroRNAs (miRNAs) are present in various body fluids and are selectively packaged inside the exosomes, a type of membrane vesicles, secreted by most cell types. These exosomal miRNAs could be actively delivered into recipient cells, and could regulate target gene expression and recipient cell function. Here, we analyzed the lactation-related miRNA expression profiles in porcine milk exosomes across the entire lactation period (newborn to 28 days after birth) by a deep sequencing. We found that immune-related miRNAs are present and enriched in breast milk exosomes (p<10(-16), χ(2) test) and are generally resistant to relatively harsh conditions. Notably, these exosomal miRNAs are present in higher numbers in the colostrums than in mature milk. It was higher in the serum of colostrum-only fed piglets compared with the mature milk-only fed piglets. These immune-related miRNA-loaded exosomes in breast milk may be transferred into the infant body via the digestive tract. These observations are a prelude to in-depth investigations of the essential roles of breast milk in the development of the infant's immune system.

Economic evaluation of treating herpes zoster with various methods of acupuncture and moxibustion.

OBJECTIVE: To analyze the cost effect of surrounding acupuncture plus electric acupuncture, cotton-sheet moxibustion, puncturing with red-hot needles, tapping plus cupping on herpes zoster. METHODS: Five hundred patients with herpes zoster were randomly divided into group A (surrounding acupuncture plus electric acupuncture), group B (cotton-sheet moxibustion), group C (puncturing with red-hot needles), group D (tapping plus cupping), and group E (Western medicine). The treatment was carried out twice a day in group E and once a day in the other four groups. The curative effect was observed on the 10th day of treatment; the cost was calculated for the five therapies, and the cost-effect ratio (C/E) and increment ratio (delta C/delta E) were analyzed. RESULTS: After the 10-day treatment, there was no statistical difference (P > 0.05) in the curative effect among the five groups. Pain being alleviated one day faster than in group E amounted to a saving of RMB 21.90 yuan in group A, a saving of RMB 21.87 yuan in group B, a saving of RMB 26.00 yuan in group C, and a saving of RMB 20.23 yuan in group D. Compared with group C, the values of delta C/delta E were RMB 1.55, 2.81, and 0.21 yuan in groups A, B, and D, respectively. CONCLUSIONS: The curative effect in groups A, B, C, and D was similar to that in group E, but the C/E was better than in group E.