Steroidal saponins and pregnane glycosides from Smilax microphylla.

Six steroidal saponins and two pregnane glycosides were isolated from the BuOH subfraction of 70% EtOH extract of Smilax microphylla C.H.Wright, among them two were new compounds (1 and 7). Pregnane glycosides were firstly isolated from the genus Smilax (Smilacaceae). Structures of the new compounds were determined on the basis of HR-ESI-MS, 1D and 2D NMR spectroscopic analysis.

[Callus cultivation and determination of gentiopicroside from Gentiana macrophylla].

OBJECTIVE: To explore and improve the utilization efficiency of Gentiana macrophylla resources. METHODS: The germination of Gentiona macrophylla seeds were used as explants to induce calluses; Cultured on various culture media and condition for callus induction and subculture by test; The content of gentiopicroside in callus was determined by HPLC. RESULTS: The optimum medium for callus was MS +2,4-D (1.0 mg/L) + 6-BA (0.5 mg/L), and MS +2,4-D (1.0 mg/L) +6-BA (0.5 mg/L) + NAA (0.5 mg/L) was the optimum for subculture,the optimum time of subculture was 30 d; The accumulated gentiopicroside reached the highest level with 0.78 mg/g DW when entered 28-30 d. CONCLUSION: The optimized culture method of callus of Gentiana. macrophylla is obtained; The growth curve of callus of Gentiona macrophylla turns on "S" appearance and the accumulated gentiopicroside increases along with the callus appreciation.

Evaluation of individualized treatment of nonproliferative diabetic retinopathy: a multicenter, randomized, parallel-controlled study.

OBJECTIVE: To evaluate the differences in the efficacy of stationary treatment and individualized treatment for patients with nonproliferative diabetic retinopathy (NPDR). METHODS: This study was a randomized, controlled, multicenter clinical trial. Participants with NPDR were randomized into the stationary treatment group or the individualized treatment group. The stationary treatment group was given the basic treatment and Qiming granules, and the individualized treatment group was given the basic treatment, Qiming granules, and individualized Chinese herbal medicines over a 12-week period. The individualized therapeutic formula was also changed over time to adjust to the changes in the clinical presentation of the patient. We conducted observations of fundus retinal exudation and hemorrhage, visual acuity, Traditional Chinese Medicine symptom scores and other indicators. RESULTS: A total of 140 participants with NPDR were randomized into the stationary treatment group or the individualized treatment group, and 132 participants completed this study. Following the 12-week treatment, significant improvements in both primary and secondary outcomes were observed in the stationary and individualized treatment groups. No remarkable difference in the primary outcomes between the two groups was observed. However, there was a significant difference in the Traditional Chinese Medicine symptom scores (18 ± 7 vs 15 ± 6; P < 0.05). There were no severe adverse effects. CONCLUSION: Compared with stationary treatment, individualized treatment is more effective at relieving the Traditional Chinese Medicine symptoms and improving vision and fundus lesions at 12 weeks post treatment.

[Study on seed morphology of the section Cruciata Gaudin].

OBJECTIVE: To provide a scientific evidence for the identification of the section Cruciata Gaudin. METHODS: Seed morphology of the section Cruciata Gaudin in different regions from Gansu were studied by means of scanning electronic microscopy (SEM). RESULTS: The seeds of section Cruciata Gaudin had relative conformability in terms of shape,size and ornamentation. But the reticulate cell of Gentiana dahurica was obviously wider than those of other species. CONCLUSION: This study provides some useful information for classification and identification of section Cruciata Gaudin.

[Study on the quality and genetic diversity of Gentiana macrophylla pall of different habitats].

OBJECTIVE: To study the quality and genetic diversity in Gentiana macrophylla of different habitats for controlling the quality of Gentiana macrophylla. METHODS: Main characteristics and microscopic identification were adapted to identify Gentiana macrophylla; HPLC method was applied for analysising the contents of Gentiana macrophylla. RAPD method was applied to study genetic structure and genetic diversity of Gentiana macrophylla. RESULTS: The experiment showed that the Gentiana macrophylla of different habitats have different main charactericstics, microscopic identifications and contents, and effective ingredient contents of gentiopicroside. The levels of population genetic diversity of various groups followed by HUANGZHONG County population (K) > HUAN County population (J) > ZiWU Mountain population (H). CONCLUSION: The study of external morphology, internal structure, active ingredient content and genetic level of Gentiana macrophylla from different areas provides a scientific basis for protecting wild species and ensuring the quality in introducing and cultivating Gentiana macrophylla.

[Karyotype analysis of Gentiana straminea].

OBJECTIVE: Study on chromosome number, karyotype of Gentiana straminea for the first time. Compare karyotypes of Gentiana straminea, Gentiana macrophylla and Gentiana dahurica. Provide cytological evidence for further studies on genetics and evolution. METHODS: Soak the root tip in 0.002 mol/L 8-oxychinolin solution for 5.7 h. Decomposed in 2.5% mixed enzyme solution for 1.6 h at 25 degrees C and use Hypotonic treatment for 3 h in refrigerator. At last, make specimen slides by the Air-drying technique. Sections combined with micrograph were used to analyze chromosome. RESULTS: The karyotypes formula of Gentiana straminea is K(2n) = 26 = 2M + 24 m, the AS. K was 52.68%, which belong to "1A" type. CONCLUSION: Compare karyotypes of Gentiana straminea, Gentiana macrophylla and Gentiana dahurica, the result showed that Gentiana macrophylla lives in highest stage of evolution. Gentiana straminea is intermediate between Gentiana macrophylla and Gentiana dahurica.

4-(2-Hydroxy-benzyl-idene)-3-methyl-isoxazol-5(4H)-one.

The mol-ecular skeleton of the title mol-ecule, C(11)H(9)NO(3), is approximately planar (r.m.s. deviation = 0.0056 Å); the two rings form a dihedral angle of 6.5 (1)°. In the crystal structure, inter-molecular O-H⋯N hydrogen bonds involving the H atom of the hydr-oxy group and the N atom of the isoxazole ring link mol-ecules into chains running along the c axis.

(Z)-4-[4-(Dimethyl-amino)benzyl-idene]-3-methyl-isoxazol-5(4H)-one.

The title compound, C(13)H(14)N(2)O(2), an isoxazol-5-one derivative, was synthesized by a one-pot, three-component condensation reaction of methyl acetoacetate, hydroxy-lamine hydro-chloride and 4-(dimethyl-amino)benzaldehyde. All the non-H atoms are co-planar [r.m.s deviation = 0.0039 Å], with a Z configuration about the C=C bond. The dihedral angle between the phenyl ring and the isoxazole ring is 2.58 (19)°.

Stationary Treatment Compared with Individualized Chinese Medicine for Type 2 Diabetes Patients with Microvascular Complications: Study Protocol for a Randomized Controlled Trial.

BACKGROUND: Microvascular complications in type 2 diabetes (T2DM), including diabatic retinopathy (DR), diabetic kidney disease (DKD), diabetic peripheral neuropathy (DPN) are the leading causes of visual loss, end-stage renal disease or amputation, while the current therapies are still unsatisfactory. Chinese medicine (CM) has been widely used for treating diabetic mellitus. However, most of the previous studies focused on the single complication. The role of CM treatment in T2DM patients with 2 or multiple microvascular complications is not clear. OBJECTIVE: To appraise the curative effect of CM in T2DM patients with 2 or multiple microvascular complications, and to compare the effects of stationary treatment and individualized treatment in T2DM patients with microvascular complications. METHODS: This trial will be an 8-center, randomized, controlled study with 8 parallel groups. A total of 432 patients will be randomized to 8 groups: DR study group (32 cases) and a corresponding control group (32 cases), DR+DKD study group (64 cases) and a corresponding control group (64 cases), DR+DPN study group (64 cases) and a corresponding control group (64 cases), DR+DKD+DPN study group (56 cases) and a corresponding control group (56 cases). The control group will receive stationary treatment, and the study group will receive individualized treatment based on CM syndrome differentiation in addition to stationary treatment. The study duration will be 50 weeks, comprising a 2-week run-in period, 24 weeks of intervention, and 24 weeks of follow-up. The outcomes will assess efficacy of treatment, improvement in CM symptoms, safety assessments, adherence to the treatment, and adverse events. CONCLUSION: This study will provide evidence of evidence-based medicine for CM treatment in two or multiple microvascular complications caused by T2DM. (Registration No. ChiCTR-IPR-15007072).

[Studies on plantlet regeneration and propagation of Gentiana stramines].

OBJECTIVE: To provide scientific basis for large scale production by studying the technique of tissue culture of Gentiana stramines. METHOD: Callus was induced from germ-free stem segment of G. stramines on a MS medium supplemented with different hormones. RESULT: The MS medium with 0.5-1.0 mg x L(-1) BA and 0.5 mg x L(-1) NAA was suitable for the induction and proliferation of cluster bads. MS medium with 1.0 mg x L(-1) IAA and 3 mg x L(-1) BA was suitable for the induction of calli. MS medium with 1.0 mg x L(-1) IAA and 2.0-3.0 mg x L(-1) BA was suitable for the subculture of calli. MS medium with 2.0 mg x L(-1) BA and 0.5 mg x L(-1) NAA was suitable for the differentiation of calli. CONCLUSION: Aseptic seeding of G. stramines can be quickly propagated by shoot culture.

Ultrasonic Elastography Research Based on a Multicenter Study: Adding Strain Ratio after 5-Point Scoring Evaluation or Not.

BACKGROUND: This study aimed to confirm whether strain ratio should be added after evaluation of lesions with 5-point elasticity scoring for differentiating benign and malignant breast lesions on ultrasonographic elastography(UE). MATERIALS AND METHODS: From June 2010 to March 2012, 1080 consecutive female patients with breast lesions were recruited into a multicenter retrospective study, which involved 8 centers across China. Each institutional ethic review board approved the study, and all the patients gave written informed consent. All the patients underwent the UE procedure and the strain ratios were calculated and the final diagnosis was made by histological findings. The sensitivity, specificity, accuracy, PPV and NPV were calculated for each of the two evaluation systems and the areas under the ROC curve were compared. RESULTS: The strain ratios of benign lesions (mean, 2.6±2.0) and malignant lesions (mean,7.9±5.8) were significantly different (p <0.01). When the cutoff point was 3.01, strain ratio method had 79.8% sensitivity, 82.8% specificity, and 81.3% accuracy, while the 5-point scoring method had 93.1% sensitivity, 73.0% specificity, and 76.8% accuracy. The areas under the ROC curve with the strain ratio method and 5-point scoring method were 0.863 and 0.865, respectively(p>0.05). The strain ratio method shows better a diagnosis performance of the lesions with elasticity score 3 and 4. CONCLUSIONS: Although the two UE methods have similar diagnostic performance, separate calculation of the strain ratios seems compulsory, especially for the large solid breast lesions and the lesions with elasticity score 3 and 4.

Could ultrasonic elastography help the diagnosis of breast cancer with the usage of sonographic BI-RADS classification?

OBJECTIVES: The purpose was to evaluate whether BI-RADS (the Breast Imaging Recording and Data System) combined with UE (ultrasound elastography) could improve the differentiation and characterization of benign and malignant breast lesions by comparing with BI-RADS. METHODS: A total of 1080 patients with 1194 breast lesions were studied retrospectively at 8 different institutions from 3 geographic areas across China (North, South, and West) from June 2010 to March 2012. Each institutional ethic review board approved the study and all patients gave written informed consent. All the cases were examined by conventional US (ultrasonography) and UE prior to ultrasound-guided core biopsy. Performance of BI-RADS and BI-RADS combined with UE were compared in different size groups, age groups and area groups. RESULTS: BI-RADS combined with UE cloud improve the accuracy by 13.2% compared to BI-RADS alone for all lesions, 23.2% for <10 mm lesions, 13.3% for ≥10-20 mm lesions, 6.3% for ≥20 mm lesions, 18.4% for <50 years group, 1.7% for ≥50 years group, 13.7% for northern area group, 17.7% for southern area group and 4.4% for western area group. CONCLUSIONS: The help which UE contributed to BI-RADS was greater for breast lesions <10mm and <50 years group.

[Primary study on measuring the internal transcribed spacer I regions of rRNA genein seeds of Gentiana dahurica].

OBJECTIVE: To amplify the PCR with the internal transcribed spacerl regions measure the base sequence of the amplified products of DNA, and to set up an identified standard on the level of molecule. METHOD: DNA from the seeds of G. dahurica was extracted by conventional method, and composed peculiar primer was used to amplify with the internal transcribed spacerl regions of the rRNA gene, and the base sequence of the amplified products by stopping the circle of the end of double deoxidation of four color fluorescent mark was measured. RESULT: It was proved by agar sugar gel electrophoresis that the PCR amplified products of the internal transcribed spacerl regions of the rRNA gene existed. The base sequence of the seeds of G. dahurica's internal transcribed spacerl regions of the rRNA gene was measured. CONCLUSION: To measure the base sequence of internal transcribed spacerl regions of the rRNA gene in the seeds of G. dahurica's is a method to identify vegetal Chinese traditional medicine on the level of molecule.

A new lyoniresinol derivative from Smilax microphylla.

A new lignan, lyoniresinol-9-O-8"-syringylglycerol ether (1), together with five known compounds, piceatannol (2), resveratrol (3), oxyresveratrol (4), quercetin-3'-glucoside (5) and diosgenin (6) were isolated from the rhizomes of Smilax microphylla. The structure of the new compound was determined by means of chemical evidence and 1D-and 2D-NMR (1H, 13C, HSQC, HMBC, 1H-1H COSY and NOESY) spectroscopic analysis and HR-ESI-MS.

Ultrasound elastography of breast lesions in chinese women: a multicenter study in China.

PURPOSE: Through analysis, the elastograms characteristics of breast lesions of Chinese women, a suitable diagnostic standard of quasistatic ultrasound elastography (UE) for Chinese women was proposed. METHODS: From June 2010 to March 2012, 1036 consecutive female patients (mean age, 44 years old) with breast lesions were recruited into a multicenter retrospective study, which involved 8 centers across China. Each institutional ethic review board approved the study, and all the patients gave written informed consent. All breast lesions underwent ultrasound and UE examination. Two radiologists analyzed the elastograms and separated the elastograms into 10 types. A final diagnosis was made on the basis of histologic findings. The characteristics of the elastograms were analyzed. Receiver operating characteristic curves were plotted for evaluating the diagnostic performance. Sensitivity, specificity, and accuracy were calculated. Differences in sensitivity, specificity, and accuracy were tested by using the McNemar test. RESULTS: There were 1150 lesions (593 benign, 557 malignant). There was a highly significant correlation between the elastogram color distribution and the percentage of malignant lesions, with a value of 0.92 (2P < .0001). Through analysis the different malignant percentages in different elastogram types, UE diagnostic standard was proposed, which was correlated with the blue percentage in the elastogram. The specificity, sensitivity, and accuracy of UE were 86.4%, 80.8%, and 83.5%, respectively. The specificity and accuracy of UE were higher than with ultrasound. The area under the curve was 0.86. CONCLUSION: UE could give valuable assessment in the diagnosis of breast lesions. The proposed UE diagnostic standard was suitable for Chinese women.

[Iron chloride for simultaneous denitrification and chemical-biological flocculation process].

The efficiency of iron chloride on simultaneous denitrification and chemical-biological flocculation process was investigated through a lab-scale study. The results showed that: there were not significant differences in the denitrification rate, COD and NO3(-) -N removal efficiency between the control reactor and the denitrification reactor with the dosage of 20 mg x L(-1) Fe after 14 days' domestication. Meanwhile the TP removal efficiency was increased to over 80%. Though sludge concentration was increased by adding FeCl3, the settling performance wasn't change because the sludge density was increased and the particle size was reduced. The sludge settling volume (SV30) and specific resistance of filtration (SRF) after adding FeCl3 were only 22% and 1.5 x 10(12) m x kg(-1), which were lower than the values in the control reactor and benefit for sludge treatment.