Comparative evaluation of four Lycium barbarum cultivars on NaIO3-induced retinal degeneration mice via multivariate statistical analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: The fruit of Lycium barbarum L. (goji berry) is a traditional Chinese medicine and is often used to improve vision. While various goji cultivars may differentially treat retinal degeneration, however their comparative effectiveness remains unclear. AIM OF THE STUDY: To evaluate the protective effects of four goji cultivars on NaIO3-induced retinal degeneration mouse model and identify the most therapeutically potent cultivar. MATERIALS AND METHODS: The principal compounds in the extracts of four goji cultivars were characterized by UPLC-Q-TOF/MS. A retinal degeneration mouse model was established via NaIO3 injection. Dark-light transition and TUNEL assays were used to assess visual function and retinal apoptosis. The levels of antioxidative, inflammatory, and angiogenic markers in serums and eyeballs were measured. Hierarchical cluster analysis, principal component analysis and partial least squares-discriminant analysis were used to objectively compare the treatment responses. RESULTS: Sixteen compounds were identified in goji berry extracts. All goji berry extracts could reverse NaIO3-induced visual impairment, retinal damage and apoptosis. The samples from the cultivar of Ningqi No.1 significantly modulated oxidative stress, inflammation, and vascular endothelial growth factor levels, which are more effectively than the other cultivars based on integrated multivariate profiling. CONCLUSION: Ningqi No.1 demonstrated a stronger protective effect on mouse retina than other goji cultivars, and is a potential variety for further research on the treatment of retinal degeneration.

[Transcriptome analysis and validation of key genes involved in biosynthesis of iridoids in Gentiana lhassica].

As the main chemical constituents, iridoids are widely distributed within Gentiana, Gentianaceae, with promising bioactivities. Based on the previous work, the transcriptome of G. lhassica, an original plant of Tibetan herb "Jieji Nabao", was sequenced and analyzed in this study, and the transcriptome databases of roots, stems, leaves, and flowers were constructed so as to explore unigenes that may encode the key enzymes in the biosynthetic pathway of iridoids. Then, qRT-PCR was used to validate the relative expression levels of 11 genes named AACT, DXS, MCS, HDS, IDI, GPPS, GES, G10H, 7-DLNGT, 7-DLGT, and SLS in roots, stems, leaves, and flowers. Also, the total contents of gentiopicroside and loganic acid were determined by HPLC, respectively. The results are as follows:(1)a total of 76 486 unigenes with an average length of 852 bp were obtained;(2)335 unigenes were involved in 19 stan-dard secondary metabolism pathways in KEGG database, with phenylpropanoid biosynthesis having the maximum number(75 unigenes), and no isoflavone biosynthetic pathway was annotated;(3)171 unigenes participatedin 27 key enzymes encoding in the biosynthetic pathway of iridoids, and 1-deoxy-D-xylulose-5-phosphate reductoisomerase(DXR) gene was highly expressed;(4)qRT-PCR results were approximately consistent with RNA-Seq data and the relative expression levels of the 11 genes were higher in the aboveground parts(stem, leaf, and flower) than in the underground part(root);(5)the total contents of gentiopicroside and loganic acid were higher in the aboveground parts(stem, leaf, and flower) than in the underground part(root), and the difference was significant. This study provides basic scientific data for accurate species identification, evaluation of germplasm resources, research on secondary pro-duct accumulation of medicinal plants within Gentianaceae, and protection of endangered alpine species.

Coupling loss characteristics of N-P-C through runoff and sediment in the hilly region of SE China under simulated rainfall.

Soil total carbon (TC), phosphorus (P), and nitrogen (N) exports from the weathered granite slopes are greatly influenced by the complex hydrological processes and terrain factors. In this study, the coupling loss characteristics of N-P-C via runoff and sediment were studied with two soil tanks under simulated rainfalls. Three soils respectively derived from the tillage layer (T-soil), laterite layer (L-soil), and sand layer (S-soil) were employed to determine the interactions of hydrology and topography on N-P-C exports under three rainfall intensities (1.5, 2.0, and 2.5 mm/min). The erosion degree of different soils displayed an order of S-soil > L-soil > T-soil. The results showed that surface flow was the main runoff form for L- and T-soil, while underground flow was predominant for S-soil. There was a linear correlation between sediment and surface flow (R2 > 0.78). Surface flow was the dominant pathway of P loss via runoff with underground flow being an important supplementation, and the main P loss pattern switched between dissolved phosphorus (DP) and particle phosphorus (PP) during the experiment. However, P lost via eroded sediment accounted for more than 94% of the TP loss amount. N presented an opposite trend to P and was mainly lost via underground flow. The main N loss form in surface and underground flow was NO3--N. Underground flow was the predominant total nitrogen (TN) loss pathway for S- and L-soil, followed by sediment and surface flow. For T-soil, TN lost via runoff was much greater than that carried by eroded sediment. TC for S-soil was mainly lost via underground flow while that for L- and T-soil was mostly lost via surface flow. Both N-P loss loads in surface flow and P loss load in underground flow were positively correlated with TC loss load (p < 0.05), indicating that the presence of organic matter brings about more nutrient losses. These results expand our understanding of the combined effects of rainfall intensity and erosion degree on runoff and sediment yields as well as N-P-C losses from the bare weathered granite slopes of SE China.

A high efficiency method combining metal chelate ionic liquid-based aqueous two-phase flotation with two-step precipitation process for bromelain purification.

This work was to develop a cost-effective and sustainable method which included metal chelate ionic liquid-based aqueous two-phase flotation (IL-based ATPF) and a two-step precipitation process for purifying bromelain from pineapple. Firstly, the metal chelate IL-based ATPF with a copper chelate-functionalized thermosensitive block copolymer (L64-IDA-Cu(II)) as trapping agent was used as the primary purification to obtain the L64-IDA-Cu(II)-bromelain complex. Secondly, the two-step precipitation process based on the thermosensitive properties of the L64-IDA-Cu(II) was mainly carried out to achieve the further purification of bromelain. According to a series of optimal experiments, the enzymatic activity recovery of final bromelain was 95.22 ±â€¯0.04%, and the purification factor reached 6.56 ±â€¯0.03. The results of recycling of ILs and the trapping agent were satisfactory. Furthermore, the conclusions of comparison with other methods proved the superiority of this method. This novel recycling purification method has a goodindustrial prospect in future.

Preparation of dendritic polymer-based magnetic carrier for application of bromelain separation and purification.

Bromelain has wide applications in different industries, such as food, textile, and medicine. Traditional approaches for bromelain separation and purification from solution still have many problems, including unsatisfactory binding efficiency, time-consuming operation, and costly equipment. In the present study, a new type of dendritic polymer-based magnetic carrier (GO@Fe3 O4 @PEI-Cu2+ ) was first prepared for bromelain separation and purification in solution. The histidine existing in bromelain could bind to Cu2+ cations adsorbed on the surface of the magnetic carrier, and the magnetic carrier showed excellent performance for bromelain separation and purification in solution, with the adsorption capacity up to 357 mg/g. The magnetic carrier also exhibited excellent property in the aspect of recyclability. It was found that the magnetic carrier also presented desirable performance for the separation and purification of bromelain from the crude extract of pineapple peel, and the bromelain structure remained intact before and after elution process. PRACTICAL APPLICATIONS: Considering many advantages of bromelain in the applications of pharmaceutical and food industries, this study is aimed at presenting a novel magnetic carrier with high stability and fabulous performance for bromelain separation and purification in solution and achieving the practical application that the magnetic carrier can efficiently separate bromelain from the crude extract of pineapple peel.

Green separation of bromelain in food sample with high retention of enzyme activity using recyclable aqueous two-phase system containing a new synthesized thermo-responsive copolymer and salt.

High cost and enzyme deactivation in purification process are the two main obstacles for the use of enzyme as green catalyst. The objective of this work was to overcome these limitations by developing a cost-effective aqueous two-phase system (ATPS) for efficient purification of enzymes with remarkable separation efficiency and high retention of enzyme activity. The ATPS was formed by thermo-responsive block copolymer PEG113-b-PNIPAM149 and salt as phase-forming components combining economy, recovery and sustainability. This strategy fabricated block copolymer with specified molecular weight and low LCST, which not only achieved better phase splitting but also ensured easy recycling for block copolymer. The developed ATPS demonstrated excellent extraction and biocompatibility for bromelain in real sample with 94.87% separation efficiency and 77.06% activity, which were remarkably higher than those obtained in EOPOEO-based ATPS. The recycling of copolymer was introduced to minimize cost, with recovery efficiency of 90% in the five cycles.

A novel OFF-ON-OFF fluorescence probe based on coumarin for Al3+ and F- detection and bioimaging in living cells.

A novel fluorescence probe L2 based on coumarin has been designed and synthesized. The probe L2 can be used for relay recognition of metal ions Al3+ and anion F- in the aqueous HEPES buffer (0.05 M, pH = 7.4), and build a OFF-ON-OFF detection system. The probe showed high selectivity and sensitivity to target ions in the process of relay recognition, and the corresponding detection limit could be as low as 0.014 µM (Al3+) and 0.03 µM (F-). Besides, the geometry optimizations of probe L2 and [L2 + Al3+] complex were carried out using the Gaussian 16 program based on DFT, and the identification mechanism of the probe was also discussed by the mass spectrometry and theoretical calculations. Moreover, the probe has also been successfully applied to detection of target ions in living cells.

Observation on the Efficacy of Acupuncture plus Intradermal Needle in Treating Mild-to-moderate Post-stroke Depression / 上海针灸杂志

Objective To observe the clinical efficacy of acupuncture plus intradermal needle in treating mild-to-moderate post-stroke depression (PSD). Method Ninty patients with mild-to-moderate PSD were randomized into group A, B and C, with 30 cases in each group. All the groups were intervened by selecting Baihui (GV20), Shenting (GV24) and Yintang (GV29). Group A received acupuncture plus intradermal needle; group B, intradermal needle alone; group C, acupuncture plus sham acupuncture. Hamilton depression rating scale (HDRS) score and Barthel index (BI) score in the three groups were observed before and at the end of the treatment, and the clinical efficacies of the three groups were compared. Result The HDRS score and BI score showed a significant change after the treatment in the three groups (P<0.01). After the treatment, the HDRS score and BI score in the group A and group B was significantly different from those in group C (P<0.05). The total effective rate was 90.0％ in group A versus 86.7％in group B, and 73.3％ in group C; there was a statistically significant difference between group C and group A or B (P<0.01, P<0.05). Conclusion Both acupuncture plus intradermal needle and intradermal needle alone are effective approaches in treating mild-to-moderate PSD. The treatment efficacy of these two methods is similar.

Integrated method of thermosensitive triblock copolymer-salt aqueous two phase extraction and dialysis membrane separation for purification of lycium barbarum polysaccharide.

A polymer-salt aqueous two-phase system (ATPS) consisting of thermosensitive copolymer ethylene-oxide-b-propylene-oxide-b-ethylene-oxide (EOPOEO) and NaH2PO4 was employed in deproteinization for lycium barbarum polysaccharide (LBP). The effects of salt type and concentration, EOPOEO concentration, amount of crude LBP solution and temperature were studied. In the primary extraction process, LBP was preferentially partitioned to the bottom (salt-rich) phase with high recovery ratio of 96.3%, while 94.4% of impurity protein was removed to the top (EOPOEO-rich) phase. Moreover, the majority of pigments could be discarded to top phase. After phase-separation, the LBP in the bottom phase was further purified by dialysis membrane to remove salt and other small molecular impurities. The purity of LBP was enhanced to 64%. Additionally, the FT-IR spectrum was used to identify LBP. EOPOEO was recovered by a temperature-induced separation, and reused in a new ATPS. An ideal extraction and recycle result were achieved.

[Modified Jujing No.2 Decoction for varicocele-induced male infertility].

Objective: To investigate the clinical effect of Jujing No.2 Decoction on male infertility induced by varicocele. METHODS: We equally randomized 70 male patients with varicocele-induced infertility into a treatment group and a control group, the former treated with modified Jujing No.2 Decoction (1 dose a day, bid) and the latter given Danshen Tablets (4 tablets once, tid) plus Liu Wei Di Huang Wan (8 pills once, tid), both for 3 months. We examined the spermatic veins of the patients by ultrasonography and performed seminal analysis before and after medication. RESULTS: Compared with the controls, the patients in the treatment group showed significant improvement after medication in sperm concentration (ï¼»12.35±2.12ï¼½ vs ï¼»18.56±4.16ï¼½ ×106/ml, P<0.05), progressively motile sperm (ï¼»16.18±6.34ï¼½ vs ï¼»24.63±8.51ï¼½ %, P<0.05), and morphologically normal sperm (ï¼»11.59±5.31ï¼½ vs ï¼»14.17±6.02ï¼½ %, P<0.05). In the control group, sperm concentration was increased from (12.29±2.07) ×106/ml at the baseline to (13.13±3.13) ×106/ml after medication, progressively motile sperm from (16.13±6.45) % to (20.55±7.30) % (P<0.05), and morphologically normal sperm from (11.62±5.35) % to (12.69±5.77) %. The total effectiveness rate was significantly higher in the treatment than in the control group (71.88% vs 46.67%, P<0.05). The inner diameter of the left spermatic vein was decreased from (0.32±0.05) mm at the baseline to ï¼»0.26±0.05ï¼½ mm after medication in the treatment group (P<0.05) and from (0.32±0.03) mm to (0.29±0.04) mm in the control (P<0.05), with statistically significant difference between the two groups after medication (P<0.05). CONCLUSIONS: Jujing No.2 Decoction is effective in the treatment of varicocele-induced male infertility.

[DNA-based identification of Gentiana robusta and related species].

The alpine plant Gentiana robusta is an endemic species to the Sino-Himalayan subregion. Also, it is one of the original plants used as traditional Tibetan medicine Jie-Ji. We sequence the nuclear ribosomal internal transcribed spacer (ITS) regions, matK, rbcL, rpoC1, trnL (UAA), psbA-trnH, atpB-rbcL, trnS( GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF( GAA) fragments of cp DNA in both G. robusta and such relative species as G. straminea, G. crassicaulis and G. waltonii. With Halenia elliptica as the outgroup, molecular systematic analysis reveals that G. robusta is a natural hybrid. G. straminea is the mother of hybrids, but the father is not very clear. In addition, the molecular markers for distinguishing G. robusta from the parental species or closely related species are identified, respectively. Our studies may provide valuable reference for the species identifications of medicinal plants with complex genetic backgrounds.

[Studies on genetic diversity of three Tibetan herbs].

The genetic diversity of three Tibetan herbs, i. e., Sang-Di, E-Dewa and Ye-Xingba (Tibetan names), was studied based on the field collection, specimen identification and DNA sequence analysis. Swertia hispidicalyx, Gentiana lhassica and Scrophularia dentata, as the original plants of the three Tibetan herbs, were collected and identified. The regions of ITS, matK, rbcL, rpoC1, trnL(UAA), psbA-trnH, atpB-rbcL, trnS (GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF(GAA) and nadl 2nd intron were amplified and sequenced. The ITS regions of S. hispidicalyx and S. dentata were cloned and sequenced, and the sequences were classified into different genotypes. All the sequences were analyzed and compared with those of closely related species. Our studies may provide reference for the genetic diversity analysis and molecular identification of the three Tibetan herbs.

[Study on Embryonic Development of Four Species of Gentiana (Gentianaceae)].

OBJECTIVE: To study the embryonic development of Gentiana straminea, G. robusta, G. crassicaulis and G. tibetica. METHODS: The seed germination rates, length and width of embryos, starch grains and chloroplasts were observed and analyzed by statistic software. RESULTS: The seed germination rates of the four species were all high. The increase of the length of embryo depended mainly on the increase of the length of hypocotyl. Starch grains were stored in cells and chloroplasts appeared in cotyledons. The process of germination was divided into eight periods. CONCLUSION: The embryo growth characteristics of the four species are recognized, and the results can be used to study the in situ conservation, genetic breeding and cultivation of Sect. Cruciata.

[In situ investigation to three dimensional structures of Chinese medicines seeds].

This paper is aimed to microscopic identification of traditional Chinese medicines (TCMs) using an in situ imaging method. In this study, two kinds of Zingiberaceae seeds, Amomi Rotundus Fructus and Alpiniae Katsumadai Semen, were investigated by synchrotron radiation in-line X-ray phase-contrast computed tomography (IXPCT) imaging method. The results showed that the microstructures of these Zingiberaceae seeds could be clearly obtained from the virtual slices information in different observing angles. It proves that IXPCT is an effective imaging method, which can provide the imaging information for the microscopic identification of the intact TCMs in situ and non-destructively.

[Multi-centered, randomized controlled clinical study on Chinese medicine formula particles for hyperlipidaemia associated with highly active antiretroviral therapy].

OBJECTIVE: To observe the effect and safety of Xiaozhi particles, integrated taohong Siwu tang and Erchen tang and Xuezhikang capsule in treating hyperlipidaemia (HLP) associated with highly active antiretroviral therapy (HAART). METHOD: In the multi-centered, randomized controlled clinical study, 180 hyperlipidaemia associated with highly active antiretroviral therapy cases were divided into the treatment group treated by Xiaozhi particles, integrated Taohong Siwu tang and Erchen tang, and the control group treated by Xuezhikang capsule. The treatment course was 12 weeks. The total cholesterol (Tch), triglyceride (TG), low density lipoprotein (LDL) and high-density lipoprotein(HDL) were observed. RESULT: After 12 weeks, compared with Xuezhikang capsule, the change difference of Tch, LDL, HDL in the Chinese traditional medicine formula groups of patients is significant (P < 0.05), the change of the TG has no significant difference. The effect of Tch, LDL in Xuezhikang capsule groups is better than in traditional Chinese medicine formula group,but the effect of HDL in traditional Chinese medicine formula group is better than in Xuezhikang capsule groups. CONCLUSION: Integrated Taohong Siwu tang and Erchen tang, Xiaozhi particles and Xuezhikang capsule can be used to control the hyperlipidaemia associated with highly active antiretroviral therapy as one of the main Chinese native medicine preparation.

[Study of traditional Chinese medicine syndrome features of AIDS-related chronic diarrhea].

OBJECTIVE: To study the AIDS-related chronic diarrhea in traditional Chinese medicine (TCM) clinical manifestations and syndrome factors, explore the characteristics of syndrome. METHOD: A multicenter, prospective collection of 311 cases of AIDS patients with chronic diarrhea, study the characteristics of TCM syndrome by using the method of descriptive statistics and exploratory factor analysis. RESULT: The common clinical manifestation of TCM: fatigue (229 cases, 73.63%), bowel (229 cases, 68.81%), diarrhea (194 cases, 62.38%), thin fur (201 cases, 64.63%), pink tongue (166 cases, 53.38%), greasy fur, thready pulse (126 cases, 40.51%), sink vein (64 cases, 20.58%), slippery pulse. 17 common factors were extracted, common disease syndrome factor as the spleen, stomach, liver, gallbladder and colon syndrome factors of disease, Qi, Yang deficiency, Qi stagnation, dampness and heat evil. CONCLUSION: AIDS-related chronic diarrhea symptoms involving multiple organs, the disease belongs to deficiency and excess.

[Identification of medicinal plants used as Tibetan traditional medicine Chuan-Bu based on nrDNA markers].

OBJECTIVE: To identify the common Tibetan herb Chuan-Bu. METHOD: Local herbalists were visited to observe which plants were being used as Chuan-Bu. Samples of the indigenous plants were collected at the same time. Leaf materials were collected from field surveys. Total genomic DNA was extracted from silica gel-dried leaf samples. The PCR products were purified and directly sequenced. RESULT: As the origin of Chuan-Bu in Tibet autonomous region was authenticated, two species were determined, i. e. Euphorbia stracheyiand E. wallichii. Also, based on our earlier research, the origin of Chuan-Bu in Gansu province, is from E. kansuensis. The sequences of ITS1 for E. stracheyi and E. wallichii were 261 bp in size, and 221 bp in ITS2, respectively. The size of the 5.8S coding region was 164 bp for all species examined in the genus. Especially, there was a heterozygous locus in ITS1 (C/G; position 72) for E. stracheyi. The nucleotide divergence between sequences of the 6 species in pairwise comparisons was calculated and the result showed that the variable site could be detected in each pairwise comparison of sequences. Also, there were 8 point mutations in the 5.8S coding region. CONCLUSION: nrDNA ITS sequences can be used as the molecular markers to identify the Tibetan herb Chuan-Bu and such Traditional Chinese Medicines from the same genus Euphorbia as E. lathyris, E. humifusa and E. pekinensis.

[Clinical feature and treatment of 69 Chinese children patients infected with influenza A (H1N1)].

OBJECTIVE: During the 2009 influenza A (H1N1) epidemic in China, children are the main group among people infected with influenza A (H1N1) virus, but few reports about children are available. The present study aimed to observe the clinical, laboratory features and to analyze therapeutic result. METHOD: The research subject were 93 children infected with influenza A (H1N1), 59 male and 34 female who were treated in Beijing Ditan Hospital from 15 May 2009 to 10 September 2009. The patients' data on symptoms, signs, chest X-ray, blood routine test, erythrocyte sedimentation rate (ESR), C reactive protein (CRP), liver function, renal function, helper T lymphocyte were collected and analyzed. The patients were treated with Oseltamivir, traditional Chinese medicine and symptomatic treatment. RESULT: The main symptoms of children infected with influenza A (H1N1) are fever (84 cases, 90.3%), cough (62 cases, 66.7%), pharyngodynia (36 cases, 38.7%) and expectoration (19 cases, 20.4%) at onset, and fever (59 cases, 63.4%), cough (52 cases, 55.9%), pharyngodynia (23 cases, 24.7%) and expectoration (9 cases, 9.7%) were the mojor symptoms and signs while the patients visited our hospital. The main signs were fervescence, pharyngeal congestion (53 cases, 57.0%), tonsilar swelling (21 cases, 22.6%), and abnormal white blood count (WBC) was found in 32 cases, abnormal ESR in 10 cases, abnormal CRP in 10 cases, abnormal CD4 T lymphocyte count in 19 cases, abnormal liver function and renal function were found in very few patients. After treatment, the febrile duration and time to virus negative in patients treated with oseltamivir alone, traditional Chinese medicine alone, combined oseltamivir and traditional Chinese medicine as well as those who were neither treated with oseltamivir nor traditional Chinese medicine were respectively 1 - 6 days (median 1 day), 3 - 13 days (median 7 days), 1 - 6 days (median 1.5 days), 4 - 11 days (median 8 days), 1 - 5 days (median 1 days), 5 - 14 days (median 8 days), 1 - 5 days (median 2 days), 4 - 13 days (median 8 days). CONCLUSION: Clinical manifestations of 93 children cases were the same as those of adults. The traditional Chinese medicine could improve symptoms of children infected with influenza A (H1N1), but other clinical therapeutic effects need further study.

Study on quality analysis of essential oil from twig and leaf of Baeckea frutescens / 中国中药杂志

<p><b>OBJECTIVE</b>To provide scientific methods for quality criterion by studying the chemical components of essential oil from Baeckea frutescens.</p><p><b>METHOD</b>The chemical components of essential oil from B. frutescens were identified by GC-MS-DS, TLC and capillary GC. The relative contents of main components were determined by area normalization.</p><p><b>RESULT</b>More than 50 peaks were separated, and 38 components were identified, which accounted for over 94% of the total GC peaks areas of the essential oil. The methods for quality evaluation of essential oil from B. frutescens by TLC and capillary GC were established.</p><p><b>CONCLUSION</b>The chemical components of essential oil from B. frutescens collected from different habitats and collecting periods have common characteristics as well as differences. Some components, such as linalool, can be used as a standard and chromatography fingerprint to analyze the quality of essential oil from B. frutescens.</p>