OBJECTIVE:To establish a method for the simultaneous contents determination of chlorogenic acid,rutin and iso-quercitrin in Morus alba,fried M. alba and honeyed M. alba. METHODS:HPLC was performed on the column of Agilent Zorbax SB-C18 with mobile phase of acetonitrile-0.2% phosphoric acid(gradient elution)at a flow rate of 1.0 ml/min,detection wavelen-gth was 350 nm ,the column temperature was 30 ℃,and injection volume was 10 μl. RESULTS:The linear range was 8.20-82.01μg/ml for chlorogenic acid (r=0.999 9),2.76-27.60 μg/ml for rutin (r=0.999 9) and 4.74-47.39 μg/ml for isoquercitrin (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 98.58%-100.91%(RSD=1.02%,n=6),99.19%-101.00%(RSD=0.82%,n=6) and 98.41%-101.51%(RSD=1.08%,n=6),respectively. CONCLU-SIONS:The method is simple with good stability and reproducibility,and can be used for the simultaneous determination of chloro-genic acid,rutin and isoquercitrin in different processed drugs of M. alba.

OBJECTIVE:To study the flavonoids in She medicine Eupatorium chinense. METHODS:Silica gel,ODS and Sep-hadex LH-20 column chromatography were conducted to isolate and purify the flavonoids in She medicine E. chinense,and com-pound structures were analyzed and identified based on the physicochemical properties and spectral data. RESULTS:From the ethyl acetate extract of E. chinense,10 flavonoids were isolated as tricin (1),quercetin(2),kaempferol(3),luteolin(4),luteo-lin-7-O-β-D-glucoside (5),4-methoxyctricin (6),quercetin-3-O-β-D-glucoside(7),kaempferol-3-O-β-D-glucoside(8),kaempfer-ol-3-O-rutinoside(9)and rutin(10). CONCLUSIONS:Compound 1-10 are isolated from E. chinense for the first time. The study provides certain basis for the quality evaluation of E. chinense.

Objective:To establish the quantitative method for eucalyptol in Chimonanthus salicifolius S. Y. Hu by GC. Methods:The GC method was performed on a Zebron ZB-WAX column (60 m × 0. 32 mm,0. 5 μm) with programmed temperature of 60-200℃, an FID detector was used, the detector temperature was 250℃, the inlet temperature was 220℃, and the carrier gas was nitrogen with high purity. Results:A good linearity of eucalyptol was within the range of 0. 012 6-0. 503 4 mg·ml-1(r=0. 999 8), and the average recov-ery was 100. 68%(RSD=1. 51%,n=6). Conclusion:The method is simple, accurate and quick with good reproducibility, and suitable for the quality control of Chimonanthus salicifolius S. Y. Hu.

Objective To construct a full-length cDNA library of human normal kidney tissues and identify the quality of the library. Methods By using the template-switching mechanism at 5′end of mRNA technique to construct the library, a powerscript reverse transcriptase was used to transcribe, and a 5′-oligo fragment as an extended template was added to 5′ end of mRNA to enrich full-length cDNAs. After amplification, the ds cDNAs digested by sfi I and size-fractionated by columns were recombined into ?TripIEx 2 vectors. After package, the recombinant vectors were titered and the recombinant rate (blue/white) was determined,then the library was amplified. We identified the library using PCR reaction to determine the size of the inserts. Results The titer of cDNA library was 2.6?10 6pfu?mL -1, the rate of recombinant was above 95%, and the titer of amplified library was 9?10 11pfu?mL -1. The insert size ranged from 0.7 to 2 kb. Conclusion The cDNA library of human normal kidne we constructed is a highly efficient one and can be used for screening by probe and antibody to find the genes related to kidney diseases.

Objective: To establish an HPLC method for the simultaneous determination of three flavonoids in Shi Liang Cha. Methods:HPLC was applied with the chromatographic conditions as follows: the chromatographic column was Agilent Zorbax SB-C18 (250 mm × 4. 6 mm, 5μm) at 30℃;acetonitrile-0. 1% H3 PO4 solution was used as the mobile phase with gradient elution; the flow rate was 1. 0 ml·min-1, and the detection wavelength was 360nm. Results: Good linearity of rutin, quercetin and kaempferol was within the range of 0.0409-1.637 0mg·ml-1(r=0.999 2), 0.44-88.00μg·ml-1(r=0.999 8) and 0.41-77.63μg·ml-1(r=0. 999 2), respectively; the average recovery was 99.35%(RSD =1. 64%),101. 14% (RSD =1. 88%) and 99. 69% (RSD =1. 92%) , respectively. Conclusion:The method is simple, accurate and repeatable, and suitable for the simultaneous determination of rutin, quercetin and kaempferol in Shi Liang Cha.

Objective To evaluate the clinical efficacy of oral Sarpogrelate hydrochloride in the treatment of chronic arterial ischemia of the lower extremities.Methods In this study 892 patients,who suffered from arteriosclerosis (ASO) or thromboangiitis obliterans ( TAO ) or diabetic foot ( DF ),with symptoms of intermittent claudication, sensation of cold, pain, ulcer, and without a history of vasotransplantation or bypass grafting or interventional therapy, were treated by taking Sarpogrelate hydrochloride tablets 100 mg tid for consecutive 8 weeks.The improvement rate of concomitant symptoms and the total effective rate of ASO, TAO, DF were evaluated.Drug adverse reaction were recorded.Results The improvement rate of intermittent claudication,sensation of cold,pain and ulcer were 96.9％,97.1％,89.0％ and 86.9％ respectively.The total effective rate for ASO,TAO,DF was 83.5％.A total of 81 cases (9.1％) reported mild side effects,including 7 patients with mild rash after 2- 5 days' medication,21 patients with mild nausea and 53 patients with stomach discomfort after 1 - 2 days' medication.Symptoms were managed conservatively without discontinuing taking sarpogrelate hydrochloride.Conclusions Sarpogrelate hydrochloride oral is a safe and effective therapy for chronic arterial ischemia diseases of the lower extremities.

BACKGROUND:In early experiments, we prepared hydroxyapatite-coated mechanical heart valve embedded with col agen using impulse laser sediment method.
OBJECTIVE:To further analyze the histocompatibility and toxicity of hydroxyapatite-coated mechanical heart valve embedded with col agen.
METHODS:After passage, canine vascular endothelial cellsuspension was inoculated onto the hydroxyapatite-coated mechanical heart valve embedded with col agen. One group was inoculated in 5%CO2, 37 ℃ incubator for 3 weeks static culture, and the other group was inoculated in 5%CO 2 , 37 ℃ incubator for 3 weeks spinner culture. Scanning electron microscope was used to observe cellattachment on the material. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to the proliferative capacity of vascular endothelial cells cultured with the hydroxyapatite-coated mechanical heart valve embedded with col agen.
RESULTS AND CONCLUSION:During the spinner culture, adherent cells were found on the surface of mechanical heart valve, and the cells distributed evenly and confluent at 21 days to cover the surface of the material. The number of adherent cells in the spinner culture was higher than that in the static culture. The cells during the static culture were aggregated and distributed irregularly. The mechanical heart valve exhibited no effects on the proliferation of canine vascular endothelial cells which grew wel . These findings indicate that the hydroxyapatite-coated mechanical heart valve embedded with col agen exert no effect on proliferation of vascular endothelial cells, has no toxicity and has good biocompatibility.

Objective To evaluate the applied value of percutaneous oxygen-ozone injection in the treatment of lumbar disc herniation under open 0.23 T MRI guidance.Methods Mounted with ipath 200 optical tracking system,MR-guided injection of oxygen-ozone were performed via a medial border of the articular processes approach in 73 patients with clinically diagnosed LDH.MR compatible 19.5G or 21.0 G biopsy needle was used. Discography was performed in order to select indication before injection oxygen- ozone into nucleus pulposus in 26 patients.Sixty-four patients were injected to three sites:(1)Six to 10 ml oxygen-ozone was injected into discs centers,injected and suctioned alternately in order to make nucleus pulposus oxidation thoroughly.(2)The needle was withdrawn according to the scale of biopsy needle and optical tracking.Then,10 ml oxygen-ozone was injected into disc herniation. (3)After that,needle was withdrawn further about 1.0—1.5 cm to outside of annulus fibrosus.Fifteen to 20 ml oxygen-ozone was injected into intervertebral foramina around nerve roots.The oxygen-ozone concentration was 35—45?g/ml. Nine patients were only performed injection of oxygen-ozone into around nerve root,while not injection oxygen-ozone to nucleus pulposus for considering bad curative effect after discography.Results All of 73 patients were successfully local targeted and treated under MRI guidance without serious complications, such as nerve root injury.After 3—6 months follow-up,total overall efficacy was 91.3% with the excellent in 28,good in 39,and poor in 6,respectively.Conclusion Open MR-guided injection of oxygen-ozone, mounted with optical tracking system,is a safe and effective minimally invasive therapy for treating LDH.

Objective To evaluate the relationship between Multi-slices spiral(MSCT) perfusion and microvessel density (MVD) in maxillofacial tumors. Methods Thirty-one cases of maxillofacial tumors were studied with MSCT perfusion imaging before operation. The time-density curve, perfusion, time to peak(TTP), and Peak enhancement imaging (PEI) of tumors were calculated. MVD of the tumors was measured with immuno-histochemical method by means of detecting factor Ⅷ in all the histologic specimens. Relativity analysis was carried between MSCT perfusion imaging parameters, perfusion curve types and MVD. Results MVD of maxillofacial tumors were higher than normal tissue. MVD remarkably correlated with malignancy of the tumors. Perfusion and time to peak (TTP) correlated well with MVD(t=7.09,4.10, P0.05). Significant difference of MVD in three types of perfusion curve was found(F=8.09,P

COVID-19 epidemic continues to spread around the world till these days, and it is urgent to develop more safe and effective new drugs. Due to the limited P3 biosafety laboratories for directly screening inhibitors of virulent viruses with high infectivity, it is necessary to develop rapid and efficient screening methods for viral proteases and other related targets. The main protease (M^pro), which plays a key role in the replication cycle of SARS-CoV-2, is highly conserved and has no homologous proteases in humans, making it an ideal target for drug development. From two different levels, namely, molecular level and cellular level, this paper summarizes the reported screening methods of SARS-CoV-2 M^pro inhibitors through a variety of representative examples, expecting to provide references for further development of SARS-CoV-2 M^pro inhibitors.