Objective To establish an HPLC-based method for the determination of α-obscurine,ferulic acid,hydroxysafflower yellow A,benzoylneaconitine,benzoylaconitine,periplosin,4-methoxysalicylsalicylate,kaempferol and oleanolic acid simultaneously in Shujin Tongluo Black Plaster.Methods HPLC method was used to determine the 9 components in Shujin Tongluo Black Plaster simultaneously.The 70%methanol extracts were analyzed using Waters SunFire C18 chromatography column(4.6 mm×250 mm,5 μm),in mobile phase containing acetonitrile-0.2%phosphoric acid solution for gradient elution with volume flow rate of 1.0 mL/min;column temperature was set at 25℃;detection wavelength was set at 230 nm.Results The 9 components had good linear relationship in their respective ranges(r≥0.999 7).The average recoveries were between 98.06%-100.56%and RSD was between 0.48%-2.56%,respectively.Conclusion The method has the advantage of repeatability,simple and fast,and can be used as the quality control of Shujin Tongluo Black Plaster.

Objective To optimize the optimal extraction process for phthalein and phenolic acid components in Angelicae Sinensis Radix-Chuanxiaong Rhizoma decoction pieces.Methods On the basis of a single factor experiment,central combination design-response surface methodology was adopted,and the extraction time,ethanol concentration,and ethanol dosage were used as influencing factors,and the total normalized values of the content of Senkyunolide I,Senkyunolide A and ligustilide,and extract yield were used as evaluation indicators to optimize the extraction process of phthalein components in Angelicae Sinensis Radix-Chuanxiaong Rhizoma decoction pieces;the total normalized values of chlorogenic acid,caffeic acid,ferulic acid,and extract yield were used as evaluation indicators to optimize the extraction process of phenolic acids components in Angelicae Sinensis Radix-Chuanxiaong Rhizoma decoction pieces.Results The optimal extraction process was to add 7 times the amount of 90%ethanol to the phthalein components,extract for 130 minutes each time,and extract twice;phenolic acid components were extracted twice with 7.5 times the amount of 65%ethanol for 120 minutes each time.Conclusion The optimized extraction process for phthalein and phenolic acids in Angelicae Sinensis Radix-Chuanxiaong Rhizoma decoction pieces is stable and feasible,which can provide a basis for subsequent research.

Objective:To optimize the extraction process of Ziyin Qinggu Pills.Methods:The solvent amount, extraction time and soaking time were taken as the influencing factors, and the overall desirability (OD) of the contents of timosaponin BⅡ, paeoniflorin, calycosin 7-O-glucoside, liquiritin and the dry extract yield were taken as the evaluation indexes, so that the optimal extraction process of Ziyin Qinggu Pills was optimally selected by the centralized composite design-response surface method.Results:The optimal extraction process of Ziyin Qinggu Pills was as follows: extraction time of 115 min, soaking time of 26 min, solvent volume of 10 times, and extraction twice.Conclusion:The optimal extraction process is reasonable, stable and reliable, which can provide the basis and foundation for the extraction process and further molding process of Ziyin Qinggu Pills.

Objective To investigate the photodynamic antitumor activity and chemical characteristics of pheophorbide A (PPBa)in vitro. Methods Breast cancer cells (MCF-7), lung cancer cells (A549), cervical cancer cells (HeLa), and three kinds of hepatoma cells (HepG2, hep3B and sk-Hep1) were planted in 96-well plates.The effects of light and dark toxicity, light intensity, and drug concentration on the phototoxicity of PPBa were investigated by MTT, and the uptake of PPBa was observed by Hoechst staining under a confocal microscope.The production of singlet oxygen was observed by flow cytometry and confocal microscopy with the reactive oxygen species detection kit.The photobleaching of PPBa was investigated by measuring the absorbance by a microplate reader according to the luminescence characteristics of PPBa. Results PPBa showed strong phototoxicity and low dark toxicity to six kinds of cancer cells, and IC₅₀ values to cancer cells were as follows: MCF-7:1.033 μmol/L, A549:1.911 μmol/L, HeLa: 2.319 μmol/L, HepG2:2.015 μmol/L, Hep3B: 2.089 μmol/L, sk-Hep1:2.467 μmol/L.The main uptake site of PPBa was the cytoplasm.The production of singlet oxygen was strongly dependent on the administration concentration, and photobleaching was very low. Conclusion For the six kinds of cancer cells, PPBa has the highest phototoxicity to breast cancer cells (MCF-7), with excellent properties and ideal photosensitizer characteristics.

Objective:To investigate the clinical characteristics and surgical treatment experience of carotid body tumor (CBT).Methods:The clinical data of 12 patients with CBT admitted to the Department of Vascular Surgery, Xuanwu Hospital, Capital Medical University from March 2013 to August 2020 were analyzed retrospectively. Among them, there were 6 males and 6 females, aged 31-83 years, with a median age of 57 years. Among the 12 patients, 2 patients were not treated surgically. The body mass index (BMI), tumor side, maximum diameter of tumor, and tumor classification, operation time, intraoperative bleeding volume, postoperative drainage volume and time, postoperative hospital stay and postoperative complications of patients undergoing surgery were recorded.Results:BMI of the 12 patients was 17.19-29.07 kg/m 2, with an average of (24.05±3.95) kg/m 2. Among the 12 patients, there were 4 tumors on the left side, 6 tumors on the right side and 2 patients had bilateral tumors. The maximum diameter of the tumor was 1.7-8.7 cm, with an average of (4.05 ± 1.89) cm. Among the 2 patients with bilateral tumors, 1 patient underwent staged resection within 9 months and 1 patient only removed the larger tumor. A total of 10 patients underwent surgical resection. All excised tumors were confirmed histopathologically to be paraganglioma. The average operation time and the amount of bleeding was(164.73 ± 74.39)min and 341.82 mL respectively. The drainage time was 1-3 d, with an average of (1.73 ± 0.65) d. The cumulative drainage volume was 22-237 mL, with an average of (77.18 ± 57.47) mL. Classification of 11 surgically resected tumors: 3 patients (3/11, 27.3%) were Shamblin Ⅰ, 7 patients (7/11, 63.6%) were Shamblin Ⅱ and 1 patient (1/11, 9.1%) were Shamblin Ⅲ. There were 1 patient of hematoma and 1 patient of acute cerebral infarction after operation. One patient with decrease in muscle strength of right limb, other surgical patients complained no complications such as stroke and cranial nerve injury when discharged. Patients undergoing surgery were hospitalized for 8-20 days, with an average of (13.36 ± 3.61) d. Conclusions:CBT is a rare paraganglioma in clinic. Surgical resection is an effective method to treat CBT. Careful operation should be carried out to avoid serious complications such as wound hematoma, cranial nerve injury and ischemic stroke.

Three-amino acid loop extension (TALE) transcription factors play important roles in plant growth and cell differentiation. There are plenty of studies on TALE transcription factors in several model plants, but not in radish (Raphanus sativas). A genome-wide bioinformatics analysis identified 33 TALE family genes in the Xiang-Ya-Bai (XYB) radish, These genes, are distributed on nine chromosomes and all contain 4-6 exons. The 33 TALE genes in radish showed a co-linearity relationship with the 17 homologous genes in Arabidopsis thaliana. Moreover, a large number of stress response cis-elements were found in the promoter regions of these genes. Expression analysis showed that four genes in the BELL subfamily were highly expressed in roots, and two genes in the KNOX subfamily were highly expressed in shoots of bolting plants and callus. All radish TALE genes contain sequences encoding the conserved HOX domain, except for the gene RSA10037940, which is homologous to Arabidopsis KNATM. The deduced 3D structures of the TALE proteins irrespective of subtypes are highly similar. All the encoded proteins were weakly acidic and hydrophilic. The radish TALE gene family is relatively evolutionarily conserved, which was consistent with results from Arabidopsis, but quite different from that of rice. This study provides important clues for studying the biological functions of TALE transcription factors in radish.
Amino Acids ; Arabidopsis/metabolism* ; Gene Expression Regulation, Plant ; Phylogeny ; Plant Proteins/metabolism* ; Raphanus/metabolism* ; Transcription Factors/metabolism*

Objective:To observe the effect of functional electrical stimulation (FES) combined with an ankle-foot orthosis (AFO) and gait training on lower limb motor function, gait parameters and the joint angles of hemiplegic stroke survivors.Methods:Thirty-two stroke survivors who met the inclusion criteria were selected and randomly divided into a control group ( n=10), an orthosis group ( n=10), and a combination therapy group ( n=12). In addition to routine medication and rehabilitation, the control group received only gait training, the orthosis group received gait training and an AFO and the combination therapy group was given FES, an AFO and gait training. All three groups were treated for four weeks. Then, the Fugl-Meyer lower extremity assessment (FMA-LE), the Brunnstrom lower extremity assessment (BRL), and Functional Ambulation Categories (FACs) were used to evaluate the lower limb motor function and walking ability of the three groups. The gait parameters of the three groups were quantified using a three-dimensional gait analyzer, and the changes in the hemiplegic gait before and after treatment were compared among the three groups. Results:After the treatment the average FMA-LE, FAC and BRL scores, time-space parameters, and joint angle parameters of all three groups had all improved significantly. After the intervention the average indicators in the combined therapy group (including stride frequency, stride length and walking speed) were all significantly better than in the other two groups.Conclusions:Adding FES to gait training with an AFO can effectively improve lower limb motor function and the walking ability of hemiplegic stroke survivors.

Objective To investigate the impact of pooling of inactivated samples on testing results of nucleic acid detection of SARS-CoV-2, and to provide a scientific detection scheme for the SARS-CoV-2 nucleic acid screening of large population samples. Methods The SARS-CoV-2 positive throat swab samples and the negative throat swab samples were inactivated at 56°C for 30 minutes, and mixed according to the ratio of positive samples to negative samples at 1:4, 1:9, and 1:19, respectively. Real-time fluorescent RT-PCR technology was used to detect the ORF-lab and N genes in the original solution and mixed solution. Results The nucleic acid test results of the 20 groups of inactivated samples were all positive. The nucleic acid test results of the 1:5 mixture and the 1:10 mixture were also positive. One group of samples of the 1:20 mixture was negative for ORF-lab and positive for the N gene. The Ct values of nucleic acid detection among all groups were significantly correlated. There was no statistically significant difference in the positive rate between different sample groups. Compared with the original solution, the Ct values of the ORF-lab gene of 1:5 mixture, 1:10 mixture, and 1:20 mixture samples increased by 1.73, 2.86, and 4.05, respectively, while the Ct values of the N gene of 1:5 mixture, 1:10 mixture and 1:20 mixture samples increased by 1.69, 2.79, and 3.25, respectively. Conclusion When conducting nucleic acid screening for SARS-CoV-2 in large population samples, a mixed test of less than 10 inactivated samples would not affect the qualitative results in most cases, but the results of weak positive samples may be affected.

OBJECTIVE： To establish the method for content determination of ligustilide and to optimize the extraction technology of volatile oil and inclusion technology in Quhan zhufeng granules. METHODS： HPLC method was adopted. The determination was performed on Waters C18 column with mobile phase consisted of methanol-water （70 ∶ 30， V/V） at the flow rate of 1 mL/min. The detection wavelength was set at 327 nm， and the column temperature was 30 ℃. The sample size was 10 μL. Using yield of volatile oil and the content of ligustilide as index， with soaking time， the amount of adding water and extraction time as factors， the extraction technology was optimized by orthogonal test. Using inclusion rate， the yield of inclusion compound and yield of volatile oil as index， with ratio of volatile oil to β-cyclodextrin， inclusion temperature and inclusion time as factors， the inclusion technology of volatile oil was optimized by orthogonal test. RESULTS： The linear range of ligustilide was 0.4-4 μg（r＝0.999 9）； RSDs of precision， stability and reproducibility tests were all lower than 2％ （n＝6）. The recoveries were 96.75％-102.03％（RSD＝2.06％，n＝6）. The optimal extraction technology of volatile oil included 10-fold water （mL/g）， soaking for 15 min， extracting for 8 h. Average yield of volatile oil was 0.310 7％， and average content of ligustilide was 0.418 0 mg/g. The optimal inclusion technology of volatile oil included ratio of β-cyclodextrin and volatile oil was 1 ∶ 8 （mL/g）； inclusion temperature was 50 ℃； inclusion time was 3 h. Average inclusion rate was 69.43％， and the yield of inclusion compound was 58.89％； the yield of volatile oil was 14.15％. CONCLUSIONS： Established determination method is simple， accurate and stable. The optimal extraction technology of volatile oil and inclusion technology are stable and feasible.

Objective To evaluate the feasibility of endovascular aortic repair (EVAR) under local anesthesia without using any contrast agent for abdominal aortic aneurysm in patients with high allergic risk to contrast agent.Methods Under local anesthesia and with no use of any contrast,percutaneous EVAR was performed in a patient with abdominal aortic aneurysm who carried high allergic risk to contrast agent.Results Percutaneous EVAR was successfully accomplished.Postoperative follow-up MRI examination showed that the abdominal aortic aneurysm was completely isolated with no endoleak.The blood flow was unobstructed in the covered stent,and bilateral renal arteries were well visualized.Conclusion For the treatment of abdominal aortic aneurysm in patients who are highly allergic to contrast agent and who have contraindications to general anesthesia,percutaneous EVAR performed under local anesthesia and using no contrast agent is safe and effective.Strict observation of indications and sufficient preoperative evaluation of clinical conditions is the key to ensure a successful operation.