Purpose　The aim is to isolate and purify the erythropoietin(EPO)and thrombomodulin(TM) from human urine.Methods　Purifying and isolating human urine with means of filtration and concentration,ion-exchange chromatography, affinity chromatography and so on.Results　4.47 mg EPO and 9.92 mg TM were obtained from 2 000 kg human urine.Their molecular weights were(38.6±1.0)kD and (60±1.4)kD erespectively, the isoelectric point, 3.60±0.02 and 3.70±0.02, yield, 31.9% and 25.0%.The purified EPO and TM was shown to be highly homogeneous by capillary zone-electrophoresis(CZE),abundant Glu，Ala and Gly include on TM. Conclusion　The high purity EPO and TM was obtained.

Objective To study the chemical constituents of Abacopteris penangiana.Methods The compounds were separated and purified by various chromatographic techniques and their structures were elucidated on the basis of physiochemical properties and spectroscopic methods.Results Seven compounds were purified and their structures were identified as:(7'Z)-3-O-(3,4-dihydroxy phenylethenyl)-caffeic acid(1),caffeiein B(2),matteucinol(3),protocatechuic acid(4),p-methoxybenzoic acid(5),β-sitosterol(6),and daucosterol(7).Conclusion Compound 1 is a new phenolic acid compound named abacopteric acid,and the other compounds are isolated from the plant for the first time.

Abstract: Objective To express and purify MPT83 protein of Mycobacterium tuberculosis and evaluate its application value in immunological diagnosis of tuberculosis (TB) using clinical samples. Methods Using Mycobacterium tuberculosis (Mtb) H37Rv genome as the template, Mtb mpt83 gene was amplified by PCR and connected to PET-21a (+) to construct prokaryotic expression vector, and then transferred into E.coli DH5α. The positive colonies were picked out and retained. The recombinant plasmid pET-mpt83 of the strain with positive colony PCR was extracted, identified by double digestion, and the samples of the positive colonies were sent for sequencing. The correctly sequenced plasmids then were transferred into BL21 competent cells for induction, expression and purification with nickel column affinity chromatography. The purified products were identified by 12 alkyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Mouse polyclonal antiserum was prepared by immunizing mice with purified protein. 8 patients clinically diagnosed as tuberculosis pleural effusion (TB group) and 8 adenocarcinomas patients (CA group) were enrolled and their pleural effusion and plasma were collected. 8 healthy people (HC group) were enrolled as the control group and their plasma were collected. An indirect ELISA was used to detect the level of specific antibodies recognizing MPT83 protein in the samples. Results Mtb MPT83 protein was successfully expressed and purified. The serum titer of MPT83 mouse polyclonal antibody was as high as 1∶1 280 000. The plasma levels of MPT83 antigen specific antibodies in TB group were significantly higher than those in HC group (P<0.05), while the plasma levels of MPT83 antigen specific antibodies in CA group were not significantly different from those in HC group (P>0.05). Compared with the HC group, there was no significant difference in pleural fluid in both the TB and CA groups (P>0.05). The ROC curve was used to analyze the OD values of plasma in TB group and HC group, and the area under the curve was greater than 0.7, showing high diagnostic efficacy. Conclusion　MPT83 protein has high antigen specificity and immunogenicity, which has great application value in the immunological diagnosis of tuberculosis.

Objective To investigate the therapeutic effect and safety of microneurosurgery associated with Gamma Knife radiosurgery on the large meningiomas in sellar region, and to clarify its clinical curative effect. Methods The clinical data of 34 patients with large meningiomas in sellar region underwent microsurgery were retrospectively analyzed.All of them underwent microsurgery,and then treated with Gamma Knife radiosurgery in one month after operation if there were residual tumors.The tumor removal of situation,complication,rate of symptom remission,and recurrence rate were analyzed.Results Among the 34 patients,total resection (7 cases of Simpson grade Ⅰ and 12 cases of Simpson grade Ⅱ)was achieved in 19 cases (55.9%),subtotal resection (Simpson grade Ⅲ )in 14 cases (41.2%), and partial resection (Simpson grade Ⅳ )was achieved in 1 case (2.9%).The major complications were cranial nerve injuries (such as oculomotor nerve, trochlear nerve and abducens nerve,n=6),the contralateral limb paresis (n = 2),postoperative bleeding (n = 1),CSF leak with infection (n=3),and secondary epilepsy (n = 3 ); no death occurred postoperatively. All these patients were followed up for about 3 to 48 months.Postoperative headache disappeared in 24 cases (80%),and 6 cases were alleviated (20%);postoperative vision improved in 12 cases (80%),remained unchanged in 2 cases (13.3%)and deteriorated in 1 case (6.7%); postoperative olfactory function improved in 5 cases (62.5%) and 3 cases unchanged (37.5%); 5 cases recoverd from the ocular motility disorder (71.4%), and 2 cases unchanged (28.6%);7 cases recoverd from thehemiplegia (100%).One of the total resection cases (5.2%)and two of the non-total resection (13.3%)suffered from tumor recurrence.All of the non-total resection cases were treated with Gamma Knife radiosurgery.Conclusion The clinical effect of microneurosurgery associated with Gamma Knife radiosurgery in treatment of large meningiomas in sellar region is satisfactory.

OBJECTIVE: To investigate the characteristics and significant of mutations of GJB2 gene, SLC26A4 gene and mitochondrial 12S rRNA in deaf children who received cochlear implantation (CI) in Yunnan and to provide the data for diagnoses and research of recovery in C1 children. METHOD: Genomic DNA was extracted from the peripheral blood samples collected from 46 children and their parents (110 cases). All the children received the CI. Their parents had normal auditory phenotype. PCR was performed and the products were sequenced by automated DNA sequencer to detect the hot spots of mutations. RESULT: The detection rates of GJB2 235delC (13.0%) and 109G>A (24.0%) mutations were significantly higher than other mutations. SLC26A was the secondary major mutation (13.0%). We found out that no patient carried the mitochondrial 12S rRNA mutations. Leukoencephalopathy, hyperbilirubinemia and hypoxic-ischemic injure were disclosed in 7 patients (15.2%). The rate of mutations in parents was 36.0% (23/64). There had no difference between Han and other racial minorities (P>0.05). CONCLUSION The CI recipients in Yunnan with a high frequency of 235 delC and 109 G>A mutation, IVS7-2A>G (6.5%) is also a common mutation related hearing loss; aminoglycoside antibiotics may not be the main reason which induced congenital deaf in CI children; environment facts was suggested to contribute another important cause. The hot-spots gene screening for the C1 children could offer an accurate genetic counseling for early diagnosis and treatment, it also provide evidences for the clinical analysis between mutations and curative effect.
Anion Transport Proteins ; genetics ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; Cochlear Implantation ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; DNA, Mitochondrial ; genetics ; Female ; Genetic Testing ; Hearing Loss ; etiology ; genetics ; rehabilitation ; Humans ; Male ; Parents ; Pedigree ; Phenotype ; RNA, Ribosomal ; genetics ; Sulfate Transporters

OBJECTIVETo investigate the correlations between p14(ARF) and E2F-1, and the role of their alterations in the tumorigenesis of the lung cancer in Gejiu and Xuanwei regions in Yunnan Province for providing the important experiment basis in revealing the molecular mechanism and looking for new markers for early diagnosis of lung cancer.

METHODSThe expression of p14(ARF) and E2F-1 was detected at theirs protein level by Immunohistochemistry S-P method in 30 specimens of lung cancer of Gejiu tin miners, 30 specimens of lung cancer of Xuanwei peasants and 20 specimens of normal lung tissue. E2F-1 mRNA was detected by ISH in 25 specimens of lung cancer of Gejiu tin miners, 25 specimens of lung cancer of Xuanwei peasants and 10 specimens of normal lung tissue. The positive signals were quantitatively analysed by HPIAS-100.

RESULTSThe positive unit (PU) of p14(ARF) and E2F-1 was 16.44 +/- 4.85 and 47.39 +/- 5.43 in Gejiu group, and 16.79 +/- 3.55 and 48.15 +/- 9.11 in Xuanwei group. Expression of p14(ARF) and E2F-1 protein in lung cancer of Gejiu and Xuanwei were statistically different compared with that in the normal lung (P < 0.01) respectively; The PU of E2F-1 mRNA was 48.58 +/- 7.75 in Gejiu group, and 49.41 +/- 8.53 in Xuanwei group, which was higher than that in normal tissue group. The differences were significant (P < 0.01). There was positive correlation between the expression of E2F-1 protein and E2F-1 mRNA in Gejiu group, Xuanwei group and normal group (P < 0.01, r = 0.833). The expression of p14(ARF) protein was significantly negatively correlated with the expression of E2F-1 protein (P < 0.01, r = -0.830).

CONCLUSIONThere is the over-expression of E2F-1 gene and the deletion of p14(ARF) gene in the tumorigenesis of the lung cancer in Gejiu and Xuanwei regions in Yunnan Province. Over-expression of E2F-1 protein in lung cancer may be caused by enhanced transcription.

China ; E2F1 Transcription Factor ; genetics ; metabolism ; Female ; Humans ; Lung Neoplasms ; metabolism ; Male ; RNA, Messenger ; genetics ; Tumor Suppressor Protein p14ARF ; metabolism

Objective:To explore the progress and prospect of evaluation methods for infertility patients.Methods:From September 2017 to December 2020,199 cases of infertility patients who have accepted mini-hysteroscopy examination in the Department of Gynecological Endocrinology,Beijing Obstetrics and Gynecology Hospital,Capital Medical University.The patients who had primary ovary insufficiency,tubal infertility were excluded;patients with hysteroscopy contradiction,such as acute or sub-acute vaginal tract inflammation,en-dometrium lesion or endometrium carcinoma,severe cardiovascular,liver or kidney insufficiency were excluded.40 patients were excluded and 15 patients lost follow up,144 patients were eventually included in the analysis.The patient's age,body mass index,infertility etiology,preoperative and postoperative stimulate ovulation cycle,pregnancy state and pregnancy time were recorded.The effect of mini-hysteroscopy on ovulation induction cycle was analyzed in pregnant patients.Results:The average age of 144 patients was 32.10±4.67 years old.96 (66.67％) patients were found to have intrauterine abnormalities,while 48 (33.33％) patients were found to have normal intrauterine cavity.48 cases of pregnancy,accounting for 33.33％;the median and interquartile ranges[M (Q1,Q3)]of ovulation induction cycles before and after surgery were respectively[3 (0,5)],[2 (1,3)],the difference was statistically significant(P ＜ 0.05).Among the 48 pregnant patients,32 of them had no structural lesions in hysteroscopy,the ovula-tion induction cycle of these patients before and after hysteroscopy were respectively[5 (0,9)],[1 (0,3)],the difference was statistically significant (P ＜ 0.05).Conclusion:Uterine cavity evaluation is important for the assessments of fertility.As an advanced diagnosis instrument,mini-hysteroscopy can not only timely detect the intrauterine abnormalities of patients,but also with great application value in shorten the ovulation cycle in infertility patients and increase the pregnancy rate.

Novel uniform-sized magnetic molecularly imprinted polymers (MMIPs) were synthesized for selective recognition of active antitumor ingredients of kaempferol (KMF) and protoapigenone (PA) in Macrothelypteris torresiana (M. torresiana) by surface molecular imprinting technique in this study. Super paramagnetic core-shell nanoparticles (γ-MPS-SiO2@Fe3O4) were used as seeds, KMF as template molecule, acrylamide (AM) as functional monomer, and N, N'-methylene bisacrylamide (BisAM) as cross-linker. The prepared MMIPs were characterized by X-ray diffraction (XRD), Fourier transform infrared spectrum (FTIR), transmission electron microscopy (TEM) and thermo-gravimetric analysis (TGA), respectively. The recognition capacity of MMIPs was 2.436 times of non-imprinted polymers. The adsorption results based on kinetics and isotherm analysis were in accordance with the pseudo-second-order model (R (2)=0.9980) and the Langmuir adsorption model (R (2)=0.9944). The value of E (6.742 kJ/mol) calculated from the Dubinin-Radushkevich isotherm model suggested that the physical adsorption via hydrogen-bonding might be predominant. The Scatchard plot showed a single line (R (2)=0.9172) and demonstrated the homogeneous recognition sites on MMIPs for KMF. The magnetic solid phase extraction (MSPE) based on MMIPs as sorbent was established for fast and selective enrichment of KMF and its structural analogue PA from the crude extract of M. torresiana and then KMF and PA were detected by HPLC-UV. The established method showed good performance and satisfactory results for real sample analysis. It also showed the feasibility of MMIPs for selective recognition of active structural analogues from complex herbal extracts.
Acrylic Resins ; chemical synthesis ; chemistry ; Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; Cyclohexanones ; chemistry ; isolation & purification ; Ferns ; chemistry ; Flavones ; chemistry ; isolation & purification ; Kaempferols ; chemistry ; isolation & purification ; Nanoparticles ; chemistry

Objective To compare the efficacy and complications of the extend endoscopic endonasal approach (EEA) and open transcranial approach for resection of craniopharyngiomas. Methods The clinical data from 46 patients with craniopharyngiomas with extend EEA and 54 patients with transcranial route in our department was analyzed retrospectively. The gross total resection (GTR) rate,length of hospital stays and complications of the two groups were compared. Results The tumor diameters of were larger in the endoscopic group than in the transcranial group (3.5 ± 1.3cm vs. 3.0±0.8 cm, P<0.05). The endoscopic group had a greater GTR rate (67.4％vs. 46.3％, P<0.05)and improved visual outcome(84.2％ vs. 59.5％,P<0.05),but lower rate of hypopituitarism (56.5％ vs. 75.9％,P<0.05)and permanent diabetes insipidus (51.4％ vs.72.7％,P<0.05). On the contrast, the endoscopic group had a greater rate of cerebrospinal fluid leak(4.3％ vs. 0.0％,P>0.05)and longer hospital stays(17.0±3.6 d vs. 13.1±2.3 d,P<0.01). Hyposmia(34.8％)and hemorrhinia (2.2％)only happened in the endoscopic group. Conclusion Compared with transcranial route, the extend EEA for craniopharyngiomas is minimal invasion and effective, which can effectively improve the GTR rate and reduce the clinical symptoms.

OBJECTIVETo compare the effects of doxycycline, losartan, and their combination in the prevention of left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in rats.

METHODSTwenty-four hours after the induction of AMI, the 254 survival rats were randomly assigned to the following groups and received drug treatment: (1) AMI controls (n=64), (2) doxycycline (30 mg x kg(-1) x d(-1), n = 63), (3) losartan (10 mg x kg(-1) x d(-1), n = 62), and (4) combination doxycycline and losartan (30 and 10 mg x kg(-1) x d(-1) respectively, n = 65) treatment groups. Also, sham operated rats (n = 30) were selected randomly. Each group was further divided into three subgroups of 1, 2, and 4 weeks of treatment. After the completion of treatment, hemodynamic studies were performed. Then, the heart of rat was fixed and analyzed pathologically.

RESULTSExclusive of the dead rats and the hearts with the myocardial infarction size < 35% or > 50%, complete experimental data were obtained in 157 rats. Besides sham operated rats, there was no significant difference in myocardial infarction sizes among the 12 subgroups of AMI control and drug treatment groups (P> 0.05). Compared with sham operated rats, left ventricular end diastolic pressure (LVEDP) and left ventricular absolute weight and relative weight (LVAW and LVRW) were significantly increased in 1, 2, and 4 week subgroups of AMI controls (P < 0.05, P < 0.01, and P < 0.001, respectively), with LVEDP elevated more significantly in 4 week than in 1 and 2 week subgroups (P < 0.01); whereas the maximum rising and dropping rate of left ventricular pressure (+/-dp/dt) and its corrected value by left ventricular systolic pressure (+/-dp/dt/LVSP) were all significantly decreased only at 4 week subgroup of AMI controls (P < 0.001). Compared with AMI controls group, LVEDP was significantly decreased in all 1, 2, and 4 week subgroups of the three treatment groups (P < 0.05, P < 0.01, and P < 0.001, respectively); LVAW and LVRW were significantly decreased in 2 and 4 week subgroups of losartan and combination groups (P < 0.05, P < 0.01, P < 0.001, respectively), and in only 4 week subgroup in doxycycline (P < 0.05, P < 0.01, and P < 0.001, respectively); whereas the maximum dropping rate of left ventricular pressure and the corrected value of left ventricular pressure rising and dropping rate were significantly increased only in 4 week subgroups of all three treatment groups (P < 0.05, P < 0.01, respectively). There is no significant difference in all indices above among the three treatment groups at all three time points (P > 0.05).

CONCLUSIONIt is indicated that doxycycline can prevent left ventricular remodeling and improve its systolic and diastolic function after AMI in rats, with the equivalent effect to that of losartan. There seems no additive effect when the two drugs are used in combination.

Angiotensin II Type 1 Receptor Blockers ; therapeutic use ; Animals ; Doxycycline ; therapeutic use ; Female ; Losartan ; therapeutic use ; Myocardial Infarction ; drug therapy ; physiopathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ventricular Remodeling ; drug effects