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 evaluate the quality of Dendrobii officinalis Caulis cultivated in Lishui by the content determination of ex-tract, polysaccharide and mannose. Methods:Totally 26 batches of Dendrobii officinalis Caulis were dried at 55℃, and the ethanol ex-tract, polysaccharide and mannose was determined respectively by hot dipping, phenol-sulphuric acid colorimetry and pre-column deri-vatization HPLC method according to the determination methods for Dendrobii officinalis Caulis recorded in Chinese Pharmacopoeia (2010 edition). Results:The contents of extract, polysaccharide and mannose in Dendrobii officinalis Caulis during the collection pe-riod were higher than those of the pharmacopoeia standard, and there were significant differences among the batches. Conclusion:The test can provide theory basis for the quality evaluation of Dendrobii officinalis Caulis cultivated in Lishui, and provide guidance for the planting of the herb.

Objective: To establish a quantitative determination method for β-eudesmol in Cortex Magnoliae Officinalis by GC. Methods:β-Phenethanol was used as the internal standard substance;the column was a Zebron ZB-WAX capillary column ( 60 m × 320 μm,0. 5μm) with the column temperature of 200℃;the detector was FID and the vaporizer temperature was 250℃; the carrier gas was nitrogen with the flow rate of 1. 3 ml · min-1 and the split ratio was 4 ∶1. Results: The linear range of β-eudesmol was 0. 015 1-0. 271 2 mg·ml-1(r=0. 999 8);the average recovery was 99. 28%(RSD =1. 17%, n=6). Conclusion:The method is simple and accurate with good reproducibility, which can be used for the quality control of medicinal materials and decoction pieces of Cortex Magnoliae Officinalis.

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 establish an HPLC method for the determination of oleanolic acid in She medicine radix of Aralia chinen-sis L. . Methods:The HPLC analysis was performed on a Waters XBridge-C18 (250 mm × 4. 6 mm, 5 μm) column. The mobile phase was methanol-0. 1 mol·L-1 ammonium acetate solution (83∶17) and the flow rate was 1. 0 ml·min-1 . The detection wavelength was 210 nm, the column temperature was 20 ℃, and the injection volume was 10 μl. Results:Oleanolic acid in radix of Aralia chinensis L. had a good separation from the other components, a good linearity was obtained within the range of 72. 52-725. 2 μg·ml-1 ( r=1. 000 0), and the average recovery was 98. 05%(RSD=1. 89%, n=6). Conclusion:The method is simple, accurate, reproducible and applicable in the assay of oleanolic acid in radix of Aralia chinensis L. .

Objective:To investigate the tumor-specific neoantigen for primary plasma cell leukemia (PCL) using gene sequencing technology combined with bioinformatic analysis. Methods: Peripheral blood samples of one patient with primary PCL during relapse and remission periods were collected. HLA molecular typing was performed using polymerase chain reaction with sequencing-based typing; whole-exome and transcriptome were sequenced by next-generation sequencing method; and bioinformatics software NetMHCpan was used to predict neoantigens. Results: Six tumor-specific missense mutations were found in the patient's peripheral blood during relapse period, located in genes FRG1, MLL3, SVIL, MYOM1, ZDHHC11 and RFPL4A.Considering patient's HLA sub-types, 43 neoantigens were predicted via bioinformatics. Considering that FRG1 and MLL3 had relatively high gene expression levels, 20 neoantigens derived from mutations of the two genes were preferentially selected, among which four neoantigens had high affinity with the patient's HLA molecules and thus had potential clinical application value. Conclusion: The study has completed a tumor neoantigen screen and prediction for primary PCL. This practice demonstrates that predicting neoantigen based on tumor-specific somatic mutation is feasible for primary PCL.

Objective To explore the potential value of tuberculosis protein chip for clinical diagnosis of tuberculosis.Methods The antibody level of tuberculosis protein ESAT-6,CFP10,16 KD,38 KD and LAM was determined in 4 093 patients,inclu-ding 441 tuberculosis and 3 652 non-tuberculosis cases by protein chip.Results The tuberculosis antibody was positive in 297 of the 441 tuberculosis cases and 647 of the 3 652 non-tuberculosis cases.Tuberculosis protein chip provided a sensitivity of 67.35% and specificity of 82.28% in the diagnosis of tuberculosis.Conclusions Tuberculosis protein chip test is a quick,easy and effective method for identifying potential tuberculosis patients with good specificity.

Objective:To analyze the positive detection rate, main genotypes of β-thalassemia in western region of Guangxi Zhuang Autonomous Region (referred to as Guangxi).Methods:Retrospective analysis of 26 189 individuals who underwent gene testing for thalassemia at the Affiliated Hospital of Youjiang Medical University for Nationalities from January 2013 to December 2019. Using the crossing breakpoint PCR (Gap-PCR) and reverse dot blot (RDB) techniques to detect Chinese common type of 7 kinds of α-thalassemia and 17 kinds of β-thalassemia genotypes, high-throughput sequencing(Sanger) was performed for suspected rare β-thalassemia. Gap-PCR was used for suspected deletion β-thalassemia types.Results:β-thalassemia was diagnosed in 4 495 (17.16%) of 26 189 samples. A total of 6 177 alleles of 20 types of β-thalassemia were detected, mainly CD17 (2 712 cases, 43.90%) and CD41-42 (2 240 cases, 36.26%), including 7 rare alleles: Gγ +( Aγδβ) 0, SEA-HPFH, Hb New York, Hb G-Taipei, Hb Hezhou, Hb G-Coushatta and IVS-Ⅱ-81. There were 3 903 case (86.83%) heterozygous, 273 case (6.07%) double heterozygous, and 319 case (7.10%) homozygous among 4 495 β-thalassaemia subjects. A total of 48 genotypes were detected. The two most common genotypes were CD17/β N (1 890 cases, 42.05%) and CD41-42/β N (1 212 cases, 26.96%), accounted for 69.01% (3 102/4 495). Seven rare genotypes were detected: Gγ +( Aγδβ) 0/β N in 3 cases, Hb New York/β N in 3 cases, Hb G-Taipei/β N in 2 cases, SEA-HPFH/β N, Hb Hezhou/β N, Hb G-Coushatta/β N and IVS-Ⅱ-81/β N in 1 case each. A total of 1 041 cases (3.97%, 1 041/26 189) of 116 types of αβ-thalassemia were detected, mainly -- SEA/αα composite CD17/β N (144 cases, 13.83%), followed by -α 3.7/αα composite CD17/β N (112 cases, 10.76%). Conclusions:Western region of Guangxi is a high prevalence area of β-thalassemia, CD17/β N and CD41-42/β N are the main genotypes. The variation spectrum of β-thalassemia is complex and diverse, with rich genotype.