1.Determination of Berberine Hydrochloride in Coptis chinensis Franch.and Xianglian Pills by Thin Layer Chromatograph-Fluorescence Spectrophotometry Method
Xinxin WEN ; Danhua LIU ; Chenhuan YU
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(10):-
Objective To establish a determining method for the content of berberine hydrochloride in Coptis chinensis Franch.and Xianglian pills.Method The sample was extracted and developed,the berberine hydrochloride was determined by thin layer chromatograph-fluorescence of spectrophotometry with Benzen-ethylaceate-isopropanol-methanol-ammonia water(6:3:1.5:1.5:0.5) as the developing system and Ex=365 nm,Em=409 nm.Result The calibration curve was linear in the range of 0.473~ 3.784 ?g,r=0.999 3.The recovery rate in Coptis chinensis Franch.was 99.03%,RSD=1.86%,and in Xianglian pills was 97.10%,RSD=1.09%.Conclusion The method is simple and accurate.
2.Treatment of Severe Meconium Aspiration Syndrome of Newborn Piglet by High Frequency Oscillatory Ventilation and Magnesium Sulphate
Fang LIU ; Shimin ZHAO ; Danhua WANG
Chinese Journal of Perinatal Medicine 1998;0(03):-
Objective To evaluate the effect of high frequency oscillation ventilation (HFOV) and HFOV+ MgSO 4 on oxygenation, pulmonary and systemic arterial pressure, histologic alterations, and serum magnesium concentration in severe meconium aspiration syndrome(MAS) piglets with persistent pulmonary hypertension(PPH). Methods Newborn piglets were randomized to HFOV group( n =6), HFOV+MgSO 4 group( n =7) and control group( n =5). Piglets of HFOV group and HFOV+MgSO 4 group received an intratracheal a 20% suspension of human meconium. All piglets were given HFOV, Piglet of HFOV+MgSO 4 group was given MgSO 4 intravenously simultaneously. Cardiopulmonary functions, blood gases were monitored and serum magnesium concentration of piglets in HFOV+MgSO 4 group were measured by atomic absorption spectrophotometric assay. Results (1) HFOV and HFOV+ MgSO 4 treatment showed improved oxygenation in MAS model, PaO 2,a/APO 2 increased significantly and A-aDO 2?,Qs/Qt decreased significantly in HFOV group at 30 minute ( P
3.Diagnosis and treatment of pheochromocytoma in pregnancy:a case report
Yingchao YANG ; Guoli LIU ; Jingwei ZHOU ; Hao HU ; Danhua SHEN
Journal of Peking University(Health Sciences) 2016;48(2):370-372
SUMMARY Pheochromocytomaisrareinpregn’ancy.Clinicalfeaturesofacaseofpheochromocytoma during pregnancy in the Peking University People’s Hospital was investigated and the literature reviewed to discuss the diagnosis and treatment of this disease.The patient manifested with hypertension and pro-teinuria,who was easily misdiagnosed with gestational hypertension disease.When she was transferred to our hospital,the symptoms such as,paroxysmal palpitation,dizziness,vomiting were noticed,and the possibility of pheochromocytoma was considered due to the accompanying abdominal mass.An emergent cesarean section was performed successfully due to preterm labor during the treatment of the disease.Af-ter the delivery the drug preparation continued.And the laparoscopic resection of pheochromocytoma pro-ceeded when the blood pressure was steady.The patient recovered fully after the surgery.The final diag-nosis of pheochromocytoma was confirmed with the pathology.Its diagnosis and treatment experiences could improve our understanding and treatment of secondary hypertension due to pheochromocytoma in pregnancy.
4.Influence of sampling satisfaction using endometrial sampling device and related factors for pathology diagnostic accordance rate
Tong ZHANG ; Rong ZHOU ; Chen LIU ; Danhua SHEN
Chinese Journal of Obstetrics and Gynecology 2014;49(9):655-658
Objective To observe the satisfaction of obtaining endometrial tissue in using endometrial sampling device and explore the influence for pathology diagnostic accordance rate,and to discuss the problem needs to focus in obtaining endometrial tissue with endometrial sampling device for endometrial cancer and precancerous lesion screening.Methods Totally 204 cases using endometrial sampling device with the subsequent diagnostic curettage,hysteroscopy,or hysterectomy were collected.A comparison of pathological diagnosis between two methods was made.The relationship among specimen quantity,proficiency,pathological types and diagnostic accordance rate were analyzed.Results The diagnostic accordance rate of endometrial cancer and precancerous lesion of two methods was 67% (20/30).92.2% (188/204) cases successfully got the endometrial glands,77.0% (157/204) cases got ≥5 specimens of glands and 62.3% (127/204) cases got ≥10 specimens of glands.When obtaining ≥5 but< 10 endometrial glands,the diagnostic accordance rate was 76.7% (23/30); however,obtaining ≥10 endometrial glands,the diagnostic accordance rate was 92.9% (118/127).The degree of specimen satisfaction of the first phase was lower than the second phase.Conclusions Obtaining ≥5 endometrial glands in using endometrial sampling device could satisfy the requirement of the basic pathological diagnosis.The proficiency of sampling and the pathological types could influence the degree of specimen satisfaction,and then influence the diagnostic accordance rate.
5.Determination of N-Acetyl-S-(N-methylcarbamoyl) cysteine in Human Urine by Online Solid Phase Extraction-High Performance Liquid Chromatography
Danhua LIU ; Hongfang TANG ; Lye LIU ; Yan JIN ; Haibao ZHU ; Zheng RUAN ; Yaling QIAN
Chinese Journal of Analytical Chemistry 2014;(12):1842-1845
A method was developed for the determination of N-acetyl-S-( N-methylcarbamoyl ) cysteine ( AMCC) in human urine by online solid-phase extraction ( SPE )-high performance liquid chromatography ( HPLC ) . The separation of AMCC from the urine matrix was performed on AmoniPac PA Solid phase Extraction ( SPE ) column with 5 mmol/L KH2 PO4 as the mobile phase by left pump. Then the time was controlled to switch the valve to make only the section of sample containing AMCC transferred into the analytic column-Acclaim PAⅡ C18 . The determination was performed using gradient elution of 0. 1% H3 PO4 (containing 5% acetonitrile) and acetonitrile by right pump. The results showed that AMCC present good linear correlation in the range of 1 . 0-100 mg/L with a correlation coefficient of above 0 . 999 , the quantitation limit of the method was 0. 2 mg/L (with the sample inject volume =10 μL), the recoveries of spiked samples were in the range of 82 . 9%-85 . 9%, and the relative standard deviation ( n=6 ) of retention time and peak area were 0. 2% and 4. 0% respectively. Compared with offline SPE-HPLC, the proposed method was convenient, environmentally friendly, efficient and stable, and feasible for the detection of AMCC in 7 human urine samples.
6.Prokaryotic expression, purification and identification of human procalcitonin
Tao FU ; Danhua SU ; Yuan LIU ; Qing LIU ; Liang WU ; Shengxia CHENG ; Xugan JIANG
Chinese Journal of Clinical Laboratory Science 2017;35(3):226-229
Objective To construct the prokaryotic expression vector of human procalcitonin(PCT) and obtain the highly purified recombinant PCT protein.Methods PCT primers were designed based on the sequence of PCT gene from NCBI,and PCT gene was amplified by PCR.Then,the recombinant vector PCT/pET-22b(+) was constructed and transferred into E.coli BL21 to induce the expression of recombinant PCT protein.Last,the recombinant PCT protein was purified by the nickel column affinity chromatography and identified by Western blot and the colloidal gold method,respectively.Results The results of agarose gel electrophoresis showed that the product of PCR amplification was about 350 bp.The homology comparison analysis revealed that the PCT gene fragment(348 bp) was successfully inserted into pTG19-T vector,and that no any base mutation was found.When the recombinant plasmid of PCT/pET-22b(+) was digested with BamH Ⅰ and HindⅢ,two pieces of about 350 bp and 5 500 bp were obtained.SDS-PAGE showed that the recombinant PCT protein with about 14 000 of Mr was existed in soluble form,and was easily obtained by the nickel column affinity chromatography.Moreover,western blot and the colloidal gold method demonstrated that the recombinant PCT protein was successfully expressed and contained histidine label(His-tag).Conclusion The recombinant vector PCT/pET-22b(+) is successfully constructed by the recombinant DNA technology,and the recombinant PCT fusion protein is successfully obtained by the nickel column affinity chromatography.
7.A study of protein expression of MAGE-A3, MAGE-A4 and MAGE-A10 genes in colorectal carcinoma and its clinical significance
Fangfang LIU ; Danhua SHEN ; Shan WANG ; Yingfiang YE ; Hui QIU ; Chenggang WANG ; Yanbin ZHANG ; Qiujing SONG
Chinese Journal of General Surgery 2012;27(1):37-39
Objective To explore protein expression and significance of MAGE genes in colorectal carcinoma(CRC)tissues.Methods The expression of MAGE genes were studied by using tissue chip and immunochemistry methods in primary CRC tissue and paired adjacent tissue samples in 97 cases.Data were analyzed with x2-test by SPSS 16.0 software.Results The protein expression of MAGE-A3,MAGE-A4 and MAGE-A10 genes were 57%(56/97),63%(61/97)and 28%(27/97)respectively in 97 cases of primary adenocarcinoma.The protein expression frequency of MAGE-A3 in poor colorectal adenocarcinomas was significantly higher than in well-and moderately disfferentiated adenocarcinomas(x2 =9.133,P =0.010).MAGE-A10 in poor colorectal primary adenocarcinomas was significantly higher than in well and moderately adenocarcinomas(x2 =15.280,P =0.000); MAGE-A10 protein expression was significantly higher in stage TNM Ⅲ + Ⅳ than in stage TNM Ⅰ + Ⅱ(x2 =4.227,P=0.040); MAGE-A10 gene expression was higher in metastasis lymphoid node than in no metastasis lymphoid node(x2 =5.557,P =0.018),and the expression level was higher in primary lesion with the increasing of the numbers of lymphoid node metastasis(x2 =7.296,P =0.026).Conclusions The protein expression of MAGE genes is associated with the tumor differentiation,TNM stage and lymphoid node metastasis.MAGE-A3 and MAGE-A10 genes are the possible prognosis marker and potential target of immunotherapy of CRC.
8.The protein expression of PLAC1/CP1 genes in colorectal carcinoma and its clinical significance
Fangfang LIU ; Danhua SHEN ; Shan WANG ; Yingjiang YE ; Chenggang WANG ; Yanbin ZHANG ; Qiujing SONG
Chinese Journal of General Surgery 2010;25(12):985-987
Objective To explore the protein expression of PLAC1/CP1 ( cancer-placenta 1 ) and its correlation with clinicopathological characteristics in patients of colorectal carcinoma. Methods The expression of PLAC1/CP1 gene was studied by using tissue chip and immunochemistry in 125 cases CRC tissue specimens. Data were analyzed with the x2-test or Fisher's x2 test statistic by SPSS 16. 0 software.Results The protein expression of PLAC1/CP1 gene was 57.6% (72/125) in 125 cases of CRC and 56. 7% (55/97)in 97 primary adenocarcinoma cases. That was 78. 9% (15/19)in poor differentiated colorectal primary adenocarcinoma, those were significantly higher than that of 35.3% (6/17) in well and 55.7% (34/61) in moderately differentiated adenocarcinoma ( P < 0.05 ); PLAC1/CP1 protein expression was significantly higher in stage TNM Ⅲ + Ⅳ 71.2% (37/52) than in stage TNM Ⅰ + Ⅱ 40% (18/45)(P < 0.05 ); PLAC1/CP1 genes expression rate was 69.6% (32/46) in these with lymphoid node metastasis and 45.1% (23/51)in patients without lymphoid node metastasis (P < 0.05 ). The positive expression rate of PLAC1/CP1 increased in colorectal carcinoma with the increasing of the numbers of lymphoid node involved with metastasis( x2 = 13. 353, P = 0.001 ). Conclusions The protein expression of PLAC1/CP1 is associated with tumor differentiation, TNM stage and lymphoid node metastasis.
9.Safety evaluation of inhaled nitric oxide in treatment of bronchopulmonary dysplasia
Qiufen WEI ; Yan LI ; Xinnian PAN ; Danhua MENG ; Xianzhi LIU ; Jing XU ; Wei WEI
Chinese Pediatric Emergency Medicine 2016;23(4):244-247
Objective To discuss the safety of inhaled nitric oxide (iNO)for bronchopulmonary dysplasia (BPD).Methods We analysed 65newborns diagnosed as BPD from January 2012to December 2013in Maternal and Child Health Hospital of the Guangxi Zhuang Autonomous Region.They were divided into iNO group(n=35)and control group(n=30)according to whether administering iNO or not.The heart rate,blood pressure,oxygen supply time,assisted ventilation time,hospital stay,concentration of NO2 and methemoglobin,and the changes of coagulation function were observed dynamically before and after iNO treatment.Results There were significant differences in oxygen supply time[(37±9)d vs.(55±7)d], assisted ventilation time[(29±7)d vs.(35±9)d],hospital stay [(45±9)d vs.(54±7)d]between iNO group and control group(P﹤0.05).There were no obvious changes in heart rate,blood pressure and co-agulation function before and after iNO treatment(P﹥0.05).The incidences of the complications had no sig-nificant differences between the two groups (P﹥0.05).Conclusion Long-time iNO can shorten oxygen therapy time and assisted ventilation time,iNO therapy in BPD doesn′t found serious adverse effect to cardio-vascular function as well as coagulation function,and doesn′t increase the risk of intracranial hemorrhage and retinopathy of prematurity.