An optimal combination of five main monomer components in Wuzi Yanzong Pill that prevents neural tube defects and reduces apoptosis and oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Wuzi Yanzong Pill (WYP) is a classic traditional Chinese medicine (TCM) formula that is used for reproductive system diseases. Previous studies showed that WYP had a preventive effect on the development of neural tube defects (NTDs) induced by all-trans retinoic acid (atRA) in mice. AIM OF THE STUDY: This study aimed to determine the optimal combination of main monomer components in WYP on preventing NTDs and to understand the underlying mechanism. MATERIALS AND METHODS: An optimal combination was made from five representative components in WYP including hyperoside, acteoside, schizandrol A, kaempferide and ellagic acid by orthogonal design method. In a mouse model of NTDs induced by intraperitoneal injection of atRA, pathological changes of neural tube tissues were observed by Hematoxylin & Eosin (HE) staining, neural tube epithelial cells apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), protein changes related to apoptosis, anti-apoptosis, and antioxidant factors were detected with Western blot. Potential targets and mechanisms of monomer compatibility group (MCG) acting on NTDs were analyzed by bioinformatics. RESULTS: Treatment with different combinations of WYP bioactive ingredients resulted in varying decreases in the incidence of NTDs in mice embryos. The combination of MCG15 (200 mg/kg of hyperoside, 100 mg/kg of acteoside, 10 mg/kg of schizandrol A, 100 mg/kg of kaempferide and 1 mg/kg of ellagic acid) showed the most significant reduction in NTD incidence. Mechanistically, MCG15 inhibited apoptosis and oxidative stress, as evidenced by reduced TUNEL-positive cells, downregulation of caspase-9, cleaved caspase-3, Bad, and Bax, and upregulation of Bcl-2, as well as decreased MDA and increased SOD, CAT, GSH, HO-1, and GPX1 levels. Bioinformatics analysis showed that MCG15 acted on the PI3K/Akt signaling pathway, which was confirmed by Western blot analysis showing increased expression of p-PI3K, p-Akt/Akt, and Nrf2 related indicators. CONCLUSION: We have identified an optimal combination of five bioactive components in WYP (MCG15) that prevented NTDs in mice embryos induced by atRA by activating the PI3K/Akt signaling pathway and inhibiting apoptosis and oxidative stress.

Purification of Polysaccharides from Orostachys cartilaginous Cell Cultures by Macroporous Resin Absorption and Bioactivities of the Purified Polysaccharides.

Orostachys cartilaginous is a traditional herbal medicine and its cell cultures contain large amounts of polysaccharides. To utilize the cultured O. cartilaginous cells, this study purified the crude polysaccharides of O. cartilaginous cells by macroporous resin absorption and optimized the purification process in the experiment of orthogonal design with four factors (sample concentration and volume, and eluent concentration and volume) and three levels; the antibacterial and anti-cancer effects of the purified polysaccharides (OTP) were further examined. The results showed that polysaccharide purity reached 95 % in the optimized group, i. e., 1.6 mg/mL of sample (crude polysaccharides) concentration, 3.0 bed volume (BV) of sample volume, 65 % eluent (ethanol) concentration, and 3.0 BV of eluent volume. In the antibacterial experiment, the growth of three bacterial species, i. e., Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis was inhibited by OTP, whereas that of Escherichia coli was not affected; the largest diameter of the inhibitory zone was found on B. subtilis; the extracellular alkaline phosphatase activity and the electrical conductivity, nucleic acid, and protein levels of B. subtilis increased after OTP treatment, indicating that the inhibition of B. subtilis growth was caused by the leakage of cell contents. In the anti-cancer experiment, OTP decreased the cell viabilities of the tested human cancer cells, i. e., AGS (gastric cancer), HCT116 (colon cancer), HepG2 (liver cancer), and HeLa (cervical cancer), and the highest inhibitory effect was on HCT116. OTP promoted HCT116 apoptosis and affected the expression of apoptosis-related proteins, i. e., the expression of B-cell lymphoma-2 decreased and that of bcl-2 associated X protein, cytochrome c, caspase 9 and caspase 3 increased. The findings of the present study suggest that O. cartilaginous cell cultures have a potential application in food or drug production.

Artificial neural network-genetic algorithm-based optimization of aerobic composting process parameters of Ganoderma lucidum residue.

The rapid development of traditional Chinese medicine enterprises has put forward higher requirements for the resource utilization of traditional Chinese medicine residues (TCMR). Aerobic composting of TCMR to prepare bio-organic fertilizer is an effective resource utilization method. In this study, a back-propagation artificial neural network (BPNN) model using composting factors as inputs (C/N, initial moisture content, type of inoculant, composting days) and the humic acid content as the output was constructed based on the orthogonal test data. BPNN-GA (a genetic algorithm) was used for extreme value optimization, and the optimal composting process parameter combination was obtained and verified. The results show that the combination of orthogonal testing and BPNN can effectively establish the relationship between the composting process parameters and humic acid content. The R2 value was 0. 9064. The optimized parameter combination is as follows: C/N,37.42; moisture content,69.76%; bacteria,no; and composting time,50 d.

An Optimal Combination of Chemically Pure Compounds from Salvia miltiorrhiza for Inhibiting Cell Proliferation.

OBJECTIVE: Salvia miltiorrhiza (SM) is a traditional Chinese medicine used clinically to treat cardiovascular diseases, including atherosclerosis and myocardial infarction. Its therapeutic effect has been confirmed by many clinical and pharmacological studies. However, the optimal formulation of active ingredients in SM for treating cardiovascular diseases remains unclear. In this study, we determined the ratio of the optimal compatibility of SM ingredients DSS, Sal-A, Sal-B, and PAL (SABP)with a uniform and orthogonal optimized experimental design. In addition, we determined the anti-oxidation effect of SABP using Adventitial Fibroblasts (AFs). METHODS: By using a combination of uniform and orthogonal designs, we determined the optimal formulation of aqueous extract from SM. MTT assay was used to determine the inhibitory effects of these 4 components of SM on the AFs, which were isolated and cultured from the aorta. The reactive oxygen species (ROS) production in AFs was compared before and after SABP treatment. RESULTS: The optimal formulation of these 4 aqueous extracts from SM were 150 : 7 : 300 : 500, and their concentrations were S(1.5×10-4 mol/L), A(7×10-6 mol/L), B(3×10-4 mol/L), and P(5×10-4 mol/L). There were some synergies between these 4 components. Moreover, SABP decreased ROS production in AFs. CONCLUSION: These findings suggest that SABP inhibits the proliferation and oxidation stress in AFs. The present study provides new evidence that the efficacy and function generated from the optimal formulation of active ingredients in SM are better than lyophilized powder of SM.

A six-herb Chinese medicine composition ointment as a promising candidate for treatment of hypertrophic scars.

Objective: To study the anti-hypertrophic scar effect of the six-herb Chinese medicine composition (SCMC) ointment on the rabbit ear hypertrophic scar models. Methods: The optimal formulation of SCMC ointment matrix was screened by the orthogonal designs and a series of evaluation tests. The SCMC ointment was prepared through emulsifying method. The rabbit ear hypertrophic scar models were established and used to investigate the anti-hypertrophic scar effect of SCMC ointment. Results: Our results demonstrated that all the quality control indications of the SCMC ointment met the requirements. Anti-hypertrophic scar activity results showed that all the rabbit ear scar tissues appeared different degrees of shrink and fading, and took an unobvious but palpable shift from hard to soft texture with the low, middle and high concentration SCMC ointments treatments in vivo. Additionally, on 21st day the scar area and thickness in different concentrations of SCMC ointment groups were significantly reduced than control group, in a concentration-dependent manner. The immunohistochemical results also indicated that the SCMC ointment had good anti-hypertrophic scar properties and could inhibit hypertrophic scar formation. Conclusion: The SCMC ointment could improve the blood circulation condition of hypertrophic scar tissues. Our research has demonstrated the Chinese medicine composition ointment with good anti-hypertrophic scar properties that could be used to treat hypertrophic scars. Meanwhile, it provides a theoretical basis for further clinical application.

A six-herb Chinese medicine composition ointment as a promising candidate for treatment of hypertrophic scars / 中草药·英文版

Objective: To study the anti-hypertrophic scar effect of the six-herb Chinese medicine composition (SCMC) ointment on the rabbit ear hypertrophic scar models. Methods: The optimal formulation of SCMC ointment matrix was screened by the orthogonal designs and a series of evaluation tests. The SCMC ointment was prepared through emulsifying method. The rabbit ear hypertrophic scar models were established and used to investigate the anti-hypertrophic scar effect of SCMC ointment. Results: Our results demonstrated that all the quality control indications of the SCMC ointment met the requirements. Anti-hypertrophic scar activity results showed that all the rabbit ear scar tissues appeared different degrees of shrink and fading, and took an unobvious but palpable shift from hard to soft texture with the low, middle and high concentration SCMC ointments treatments in vivo. Additionally, on 21st day the scar area and thickness in different concentrations of SCMC ointment groups were significantly reduced than control group, in a concentration-dependent manner. The immunohistochemical results also indicated that the SCMC ointment had good anti-hypertrophic scar properties and could inhibit hypertrophic scar formation. Conclusion: The SCMC ointment could improve the blood circulation condition of hypertrophic scar tissues. Our research has demonstrated the Chinese medicine composition ointment with good anti-hypertrophic scar properties that could be used to treat hypertrophic scars. Meanwhile, it provides a theoretical basis for further clinical application.

Optimization and mechanism of postponing aging of polysaccharides from Chinese herbal medicine formula.

To study the extraction technology of polysaccharides (AAP) from Chinese herbal medicine formula and its mechanism of delaying aging. First, L9(3)4 orthogonal test was used to optimize the optimal enzyme-assisted extraction parameters of polysaccharides. And the anti-aging effects was evaluated by detecting mitochondrial function, protein, DNA, adhesion molecules and cell cycle in aging rats. The optimal extraction process parameters were the cellulase concentration of 1.5%, the pH at 5, the enzyme temperature at 50°C and the extraction time of 180 min. The anti-aging results showed that AAP can effectively increase the activities of malate dehydrogenase, succinate dehydrogenase and superoxide dismutase. It also can decrease the activity of monoamine oxidase and methane dicarboxylic aldehyde levels in the brain tissue. Meanwhile, the polysaccharides enhanced telomerase activity while reduced p16 protein expression of the brain mitochondria. In addition, the polysaccharides continued to improve heart damage and significantly lessen mitochondrial DNA concentrations. For a certain period of time, it also enhanced the activity of superoxide dismutase, reduced glutathione, glutathione peroxidase and decreased protein carbonyl and methane dicarboxylic aldehyde content of kidney in D-galactose-induced aging rats. Furthermore, the polysaccharides restored the number of cells in the peripheral blood lines and BMNC through inhibiting the drop of the number of red blood cells, white blood cells, platelets in the peripheral blood and bone marrow mononuclear cell of the aging rats. At the same time, AAP accelerated G1 phase cell to enter S phase in cell cycle in aging rats. Our research suggests that the polysaccharides may be a potential anti-aging agent and can be further developed as a functional food or new drug to delay aging or treat aging-related diseases.

Trace water activity could improve the formation of 1,3-oleic-2-medium chain-rich triacylglycerols by promoting acyl migration in the lipase RM IM catalyzed interesterification.

New structured lipids with 1,3-oleic-2-medium chain (OMO) triacylglycerols were synthesized by promoting acyl migration in Lipozyme RM IM catalyzed interesterification between coconut oil (CO) and high oleic rapeseed oil (HORO). Results from an orthogonal design L25(55) showed that the maximal yield of OMO-structured triacylglycerols was 45.65% under the following conditions: the molar ratio of CO to HORO, 50:50; enzyme dosage, 12 wt%; reaction temperature, 60 °C; reaction time, 2 h; water activity, 0.07. Low water activity showed a high rate of acyl migration (10.86% vs 5.07% no water system), which promoted OMO synthesis due to medium-chain fatty acid migration to the sn-2 position. In a low water content (5%) system of the molecular dynamics simulation, water molecules stabilized the whole structure of RM IM through hydrogen bonding, which helped fix lipase-catalyzed active sites, making substrates more easily inserted into active sites, resulting in increased enzyme activity.

Manipulation parameter optimization in Liu's back tuina therapy for kids' cough variant asthma in remission stage / 针灸推拿医学（英文版）

Objective: To determine the optimal manipulation parameters in Liu's back tuina therapy for remission-stage cough variant asthma (CVA) in kids. Methods: A 5-factor 3-level L18(37) orthogonal design was adopted. The experimental factors and levels were selected: A, the number of times of Rou-kneading Feishu (BL 13); B, the frequency of Rou-kneading Feishu (BL 13); C, the number of times of Tui-pushing in a '介'-like pattern; D, the frequency of Tui-pushing in a '介'-like pattern; E, the intensity of red color after Ca-scrubbing Feishu (BL 13) with salt. A1, A2 and A3 stood for 10 times, 20 times and 30 times, respectively; B1, B2 and B3 stood for 200 times/min, 240 times/min and 280 times/min, respectively; C1, C2 and C3 stood for 50 times, 75 times and 100 times, respectively; D1, D2 and D3 stood for 150 times/min, 175 times/min and 200 times/min, respectively; E1, E2 and E3 stood for pinkish, bright red and dark red, respectively. A total of 108 kids with CVA in remission stage were divided into 18 groups using the random number table method, with 6 cases in each group. Changes in the cough score and fractional exhaled nitric oxide (FENO) level were observed after 15-day treatment. Results: The cough score and FENO level changed significantly in every group after treatment (all P<0.01). By the effect in improving cough score, the factors were listed as D, B, A, C and E in descending order; the most optimal manipulation parameter combination was group 17 (A3B2C1D3E1); the second optimal parameter combinations were group 7 (A3B1C2D1E3), group 13 (A2B1C2D3E1) and group 14 (A2B2C3D1E2). Regarding the effect in down-regulating the level of FENO, the factors were listed as C, A, E, B and D in descending order; the most optimal manipulation parameter combination was group 13 (A2B1C2D3E1); the second optimal parameter combinations were group 18 (A3B3C2D1E2), group 16 (A3B1C3D2E3) and group 14 (A2B2C3D1E2).Conclusion: Two most effective manipulation parameter combinations were selected out of 18 different parameter combinations in Liu's back tuina therapy for remission-stage CVA in kids. Doctors can choose an appropriate manipulation based on the symptom features of the disease.

[Study on preparation of volatile oil from Acorus tatarinowii self-nanoemulsion dropping pills and its protective effect on acute myocardial ischemia injury].

In this study, solid dispersion technology was used to develop volatile oil from Acorus tatarinowii self-nanoemulsion dropping pills(VOA-SNEDDS-DP) and its protective effect on acute myocardial ischemia injury was evaluated. Taking exterior quality, weight variation and the resolving time as comprehendsive evaluation indexes, the preparation process and formulation of the dropping pills were optimized by orthogonal design, and the dissolution rate in vitro of the optimized VOA-SNEDDS-DP was investigated. The rat model of acute myocardial ischemia was induced by intraperitoneal injection of isoproterenol hydrochloride and the serum levels of superoxide dismutase(SOD), malondialdehyde(MDA), creatine kinase(CK) and pathological changes of myocardial tissue were determined to evaluate therapeutic effect of the dropping pills on acute myocardial ischemia. The results showed that the optimal formulation and preparation process of VOA-SNEDDS-DP were as follows: PEG6000-PEG8000 was 1∶1, proportion of VOA-SNEDDS and matrix was l∶2.5, the temperature of drug fluids was 75 ℃, drop rate was 35 drops/min, drop distance was 5 cm, the condensing agent temperature was 2-10 ℃. The content of ß-asarone in the dropping pills was 42.46 mg·g~(-1). The accumulated dissolution rate of the dropping pills reached 93.85% in 10 min. The results of pharmacodynamic experiments showed that VOA-SNEDDS-DP could significantly increase the SOD content(P<0.05), reduce the levels of MDA and CK(P<0.05) in serum, and effectively improve the pathological morphology of myocardial tissue. These results revealed that the preparation of VOA-SNEDDS-DP by solid dispersion technology was stable and feasible, and VOA-SNEDDS-DP had protective effect on acute myocardial ischemia injury.

Study on preparation of volatile oil from Acorus tatarinowii self-nanoemulsion dropping pills and its protective effect on acute myocardial ischemia injury / 中国中药杂志

In this study, solid dispersion technology was used to develop volatile oil from Acorus tatarinowii self-nanoemulsion dropping pills(VOA-SNEDDS-DP) and its protective effect on acute myocardial ischemia injury was evaluated. Taking exterior quality, weight variation and the resolving time as comprehendsive evaluation indexes, the preparation process and formulation of the dropping pills were optimized by orthogonal design, and the dissolution rate in vitro of the optimized VOA-SNEDDS-DP was investigated. The rat model of acute myocardial ischemia was induced by intraperitoneal injection of isoproterenol hydrochloride and the serum levels of superoxide dismutase(SOD), malondialdehyde(MDA), creatine kinase(CK) and pathological changes of myocardial tissue were determined to evaluate therapeutic effect of the dropping pills on acute myocardial ischemia. The results showed that the optimal formulation and preparation process of VOA-SNEDDS-DP were as follows: PEG6000-PEG8000 was 1∶1, proportion of VOA-SNEDDS and matrix was l∶2.5, the temperature of drug fluids was 75 ℃, drop rate was 35 drops/min, drop distance was 5 cm, the condensing agent temperature was 2-10 ℃. The content of β-asarone in the dropping pills was 42.46 mg·g~(-1). The accumulated dissolution rate of the dropping pills reached 93.85% in 10 min. The results of pharmacodynamic experiments showed that VOA-SNEDDS-DP could significantly increase the SOD content(P<0.05), reduce the levels of MDA and CK(P<0.05) in serum, and effectively improve the pathological morphology of myocardial tissue. These results revealed that the preparation of VOA-SNEDDS-DP by solid dispersion technology was stable and feasible, and VOA-SNEDDS-DP had protective effect on acute myocardial ischemia injury.

Pain management of surgical abortion using transcutaneous acupoint electrical stimulation: An orthogonal prospective study.

AIM: This study aimed to evaluate the optimal combination of parameters for the management of pain during surgical abortion using transcutaneous acupoint electrical stimulation (TEAS). METHODS: This study recruited patients scheduled for surgical abortion between October 2014 and August 2015. The treatment protocol was created using three levels for each factor (stimulating time, acupoints, age, and parity). The primary outcomes were intraoperative visual analog scale (VAS), postoperative VAS, cervical relaxation degree and intraoperative blood loss. The secondary outcomes were the vital signs. RESULTS: Stimulation time was associated with intraoperative VAS scores (P < 0.001), acupoints were associated with postoperative VAS scores (P = 0.037), and age was associated with postoperative VAS scores (P < 0.043). Parity (P = 0.025) was associated with heart rate. A comprehensive analysis of the parameters revealed the best levels for each (stimulation time: from 15 min before operation to immediate postoperative; acupoints: SP 6 and LR 3; patient age 25.1-30.0 years; and parity: G≥2 P0 A≥1 ). Seven patients did not complete follow-up. The remaining 135 subjects did not show continuous vaginal bleeding, abdominal pain, fever or any other adverse effect. CONCLUSION: During surgical abortion, TEAS stimulation from 15 min before operation to immediate postoperative, SP 6 and LR 3, age 25.1-30.0 years and G≥2 P0 A≥1 were associated with the best analgesic effect.

[Study of optimal parameters of scalp electroacupuncture for rehabilitation effect on children of cerebral palsy].

OBJECTIVE: To analyze the effect difference of wave, intensity, time and treatment frequency by orthogonal design so as to explore the optimal parameters of scalp electroacupuncture (EA) for rehabilitation effect on children of cerebral palsy. METHODS: Ninety children of cerebral palsy were assigned into 9 groups by orthogonal design, 10 cases in each one. The acupoints were bilateral excitable area, foot motor sensory area, speech two area, speech three area, balance area, and intelligent nine acupoints, including Shenting (GV 24), Sishencong (EX-HN 1), and bilateral Benshen (GB 13) and Touwei (ST 8). EA was applied at bilateral excitable area and speech three area. We designed an orthogonal experiment with four factors and three levels. We studied wave (sparse wave of 2 Hz, density wave of 100 Hz, sparse and density wave of 2 Hz /100 Hz), intensity (1 mA, 2 mA, intensity based on tolerance), time (10 min, 20 min, 30 min), frequency (once a day, once every other day, twice a week). The Gesell developmental scale was used to evaluate the developmental quotient (DQ); and gross motor function measure (GMFM), motor function before and after treatment. RESULTS: The optimal parameters for DQ and GMFM were 2 Hz/100 Hz, 20 min, once every other day. . CONCLUSION: The optimal parameters for cerebral palsy may be 2 Hz/100 Hz, 20 min, once every other day.

Preliminary Optimization of Acupuncture Protocol in Treating Migraine in Acute Stage / 上海针灸杂志

Objective To preliminarily optimize the acupuncture protocol in treating migraine in acute stage. Method Ninety patients with migraine in acute stage were observed, with Visual Analogue Scale (VAS) as the evaluation index and an orthogonal design. Acupoints groups [Taiyang (EX-HN5), Fengchi (GB20), Taiyang (EX-HN5) and Fengchi (GB20)], insertion directions (perpendicular, downward penetration, and backward penetration), stimulation dosage (1 needle, 3 needles, and 5 needles) and acupuncture duration (30 min, 1 h, and 2 h), altogether 4 factors and 3 levels, formed up different acupuncture protocols to observe the analgesic efficacy in treating migraine in acute stage, so as to determine the role of the four factors (Chi-square test), advantage of the 3 levels (multiple comparisons) and the optimal grouping of the 4 factors and 3 levels. Result Acupoints group, insertion direction and stimulation dosage were the major factors in acupuncture analgesia, and the acupuncture duration was the secondary factor (P<0.05). The analgesic effect of Taiyang (EX-HN5) and Fengchi (GB20) was more significant than either Taiyang (EX-HN5) or Fengchi (GB20) (P<0.05). Penetration puncture from Naokong (GB19) towards Fengchi (GB20) (downward penetration) produced a more significant analgesic effect than from Fengchi (GB20) towards Wangu (GB12) and from Tianyou (TE16) towards Fengchi (GB20) (both backward penetration) (P<0.05). Acupuncture with Five needles and 3 needles (parallel horizontal insertion) were superior to that with 1 needle (P<0.05); acupuncture with 5 needles was better than that with 3 needles but without a statistical significance (P>0.05). Needle retaining for 2 h produced a better analgesic effect than retaining for 30 min and 1 h, but without statistical significances (P>0.05). Conclusion Taiyang (EX-HN5) plus Fengchi (GB20), downward penetration acupuncture, 5-needle parallel horizontal acupuncture, and 2-h needle retaining combine an optimal acupuncture protocol in treating migraine in acute stage. Nevertheless, 3-needle parallel acupuncture with 30-60-min needle retaining can also be chosen according to the condition of the patients.

Analysis and identification of volatile components of different parts of Clausena lansium in Hainan by HS-SPME-GC-MS / 中草药

Objective A headspace-solid phase microextraction-gas chromatography-mass spectrometry method (HS-SPME-GC-MS) was adopted for analyzing the volatile components of different parts of Clausena lansium in Hainan Province. Methods Five different fibers were investigated and optimized. Other five experimental parameters such as volume of water, extraction temperature, equilibrium time, extraction time, and salt concentration had been evaluated and optimized by means of the orthogonal design with L16(45) table. Finally, the volatile components of C. lansium leaves, pericarps, and seeds in Hainan were analyzed and identified by GC-MS combined with retention index (RI). Results The optimum extraction conditions were as follows: a 50/30 μm DVB/CAR/PDMS fiber, 10 mg sample powders, 2.0 mL water, 0.2 g NaCl, extraction temperature 80 ℃, equilibrium time 30 min, extraction time 60 min. A total of 83 chemical components were identified from leaves, 96 from pericarps, and 106 from seeds, representing the relative contents of 95.24%, 92.15%, and 95.92% of the total composition. The highest contents were sesquiterpenes in all of the parts, but there were obviously different both in components and contents. Conclusion The HS-SPME-GC-MS method is rapid and sensitive, with a small sample size, without any organic solvents. GC-MS combined with RI has improved the accuracy of analysis and identification. The results may provide experimental basis for further exploitation of C. lansium in Hainan. This method can be used to perform enrichment analysis of the components with high-boiling point and micro-components, which can comprehensively and scientifically characterize and evaluate the quality of Chinese materia medica.

Study of optimal parameters of scalp electroacupuncture for rehabilitation effect on children of cerebral palsy / 中国针灸

<p><b>OBJECTIVE</b>To analyze the effect difference of wave, intensity, time and treatment frequency by orthogonal design so as to explore the optimal parameters of scalp electroacupuncture (EA) for rehabilitation effect on children of cerebral palsy.</p><p><b>METHODS</b>Ninety children of cerebral palsy were assigned into 9 groups by orthogonal design, 10 cases in each one. The acupoints were bilateral excitable area, foot motor sensory area, speech two area, speech three area, balance area, and intelligent nine acupoints, including Shenting (GV 24), Sishencong (EX-HN 1), and bilateral Benshen (GB 13) and Touwei (ST 8). EA was applied at bilateral excitable area and speech three area. We designed an orthogonal experiment with four factors and three levels. We studied wave (sparse wave of 2 Hz, density wave of 100 Hz, sparse and density wave of 2 Hz /100 Hz), intensity (1 mA, 2 mA, intensity based on tolerance), time (10 min, 20 min, 30 min), frequency (once a day, once every other day, twice a week). The Gesell developmental scale was used to evaluate the developmental quotient (DQ); and gross motor function measure (GMFM), motor function before and after treatment.</p><p><b>RESULTS</b>The optimal parameters for DQ and GMFM were 2 Hz/100 Hz, 20 min, once every other day. .</p><p><b>CONCLUSION</b>The optimal parameters for cerebral palsy may be 2 Hz/100 Hz, 20 min, once every other day.</p>

[Optimization of acupoint application scheme in the treatment of bronchial asthma based on the orthogonal design method].

OBJECTIVE: To study the primary and secondary factors of the allergic history, the frequency of acupoint application and the time of acupoint application in the treatment of bronchial asthma and optimize its scheme. METHODS: Eighty patients of bronchial asthma were selected as the subjects in the orthogonal trial. The herbal medicines were the empirical formula of acupoint application (prepared at the ratio as 2:2:1:1:1:1:1:1:1 of semen brassicae, rhizome corydalis, unprocessed radix kansui, asarum sieboldii, ephedra, semen lepidii, syzygium aromaticum, cortex cinnamomi and fructus gleditsiae) and used on bilateral Feishu (BL 13), Xinshu (BL 15), Geshu (BL 17) and Shenshu (BL 23). Firstly, two groups were divided according to allergic history (40 cases with allergic history and 40 cases without allergic history), and then four subgroups were divided on the basis of the two main groups, 10 cases in each one. Through studying three factors and two levels, i.e. allergic history (Factor A:AⅠ:with allergic history; AⅡ:without allergic history), the frequency of acupoint application (Factor B:BⅠ:4 times; BⅡ:10 times, in which, in the group of 4-time applications, the application was given once every 10 days; in the group of 10-time applications, the application was given once every 4 days); and the time of application (Factor C:CⅠ:4 h; CⅡ:8 h), the optimal scheme was screened on the basis of the attack frequency before and after treatment and the score of the asthma quality life questionnaire (AQLQ) before treatment and 6 months after treatment in the patients of each group. RESULTS: ① The orthogonal trial indicated that the best optimal scheme was AⅠBⅡCⅠ, meaning the patients with allergic history were treated with acupoint application for 10 times, remained for 4 h. ②Factor B (frequency of acupoint application) and C (time of acpoint application) were the significant influential factors of AQLQ scores (both P<0.05). ③The comparison of the attack frequency and AQLQ score before and after treatment in all of the patients showed that the different combinations of factor levels induced the different impacts on the asthma attack frequency and AQLQ scores. Except in the group No.1 and the group No.5, the improvements were all significant in the rest groups, indicating the significant differences (P<0.05, P<0.01). CONCLUSIONS: Acupoint application reduces apparently the attack frequency of asthma in the patients and improves the living quality. The primary and secondary relationship among the allergic history, the frequency of acupoint application and the time of acupoint application for the impacts on the therapeutic effects are:the frequency of acupoint application > the time of acupoint application > the allergic history. The best optimal scheme is AⅠBⅡCⅠ, meaning the patients with allergic history are treated with acupoint application for 10 times, remained for 4h.

[Optimize aqueous extract protocol for Yugubao formula by orthogonal experiment design].

To determine the optimum aqueous extract protocol for Yugubao traditional Chinese medicines formula by using orthogonal experiment design. Through serum pharmacology research, L9(34) orthogonal design with single factor investigation was used to optimize the aqueous extract protocol for Yugubao formula. The effect of water extraction on activity of alkaline phosphatase (ALP) in osteoblast was referred as the evaluation index for investigating four factors: water consumption (A), heating time (B), soaking time (C), and number of decocting (D), analyzing the optimum extraction conditions, and verifying the effectiveness of this process. The optimum aqueous extract protocol for Yugubao was as follows: adding 8 times water into Chinese medical materials, heating for 60 min, soaking for 30 min, and decocting for 1 time. The drug serum of this aqueous extract of Yugubao could significantly up-regulate the osteogenic genes expression. The optimum aqueous extract protocol for Yugubao formula was established in this experiment, providing evidence for the development and utilization of Yugubao traditional Chinese medicines formula.

Clinical Study on Acupuncture in Intervening Hypermyotonia of the Affected Limbs in Stroke Based on Orthogonal Experimental Design / 上海针灸杂志

Objective To explore the optimal acupuncture scheme in intervening hypermyotonia of the affected limbs in stroke.Method Eighty eligible stroke patients were randomly grouped by using the orthogonal experimental design, by adopting electroacupuncture (A), daily acupuncture frequency (B), and acupoint (C) as three factors, and two levels. A total of 20 treatment sessions were conducted. Each group was scored by using the Modified Ashworth Scale (MAS) before and after the treatment, and the safety was also evaluated.Result There was a significant difference in comparing the MAS score between the two different levels of factor A (P<0.05), while there were no significant differences in comparing the MAS scores between different levels of factor B and factor C (P<0.05). Concerning the improvement of the MAS scores of wrist, elbow, knee, and ankle joints, A2B1C1(i.e. electroacupuncture twice a day with acupoints from yin meridians) was the optimal treatment scheme.Conclusion Electroacupuncture twice a day is the optimal treatment scheme for hypermyotonia of the affected limbs in stroke, as it can effectively ease the hypermyotonia, improve the function and symptoms of the limbs, and has a satisfactory security evaluation.

Study on process optimization and judgment standard of quantitative of cooked Rhei Radix et Rhizoma / 中草药

Objective: To optimize the processing technology of cooked Rhei Radix et Rhizoma (RRR), and to quantify the judgment standard of concocting end point. Methods: The optimum technology of combination of single factor and orthogonal design was used to determine the optimum technology of aloe-emodin, rhein, emodin, chrysophanol, emodin, and total anthraquinone. The color value of processed products was quantified using the color difference measurement. Results: The steaming time had significant effect on the content of total anthraquinone in RRR, but the steaming time and steaming temperature had no statistical significance. The values of L (4 mm), a (4 mm, 8 mm), and b (4 mm) were significantly positive (negative) with chrysophanol, emodin, emodin, aloe-emodin, and total anthraquinone. The results showed that there was a significant positive correlation between rhein and b (8 mm) values, and the linear regression equation was Y = 0.288 + 0.007 X. Conclusion: The best processing technology of cooked RRR is: RRR and 30% yellow wine were moisten for 3.5 h, set at 100 ℃ steam for 1.5 h. Concocted the end point of cooked RRR processing scientifically determine that can be determined by the L, a, and b color values. And also for the in-depth study of traditional Chinese medicine concocted the end to determine the method laid the foundation.