1.Cerebral microbleeds and intracerebral hemorrhage
Baocheng WANG ; Xiaodong JIANG ; Kaimin HU ; Tiecheng LANG
International Journal of Cerebrovascular Diseases 2010;18(6):423-426
The prognosis in patients with cerebral hemorrhage is poor. Studies have shown that cerebral microbleeds are closely related with intracerebral hemorrhage. Cerebral microbleeds may be a precursor to the occurrence or recurrence of intracerebral hemorrhage. This article reviews the relationship between cerebral microbleeds and intracerebral hemorrhage from the aspects of epidemiology, etiology, pathophysiology, clinical observation, and complications of cerebral hemorrhage.
2.Construction of tumor targeting ScFv library and screening of tumor vessel-specific antibody by phage display in vivo
Xi QIN ; Yuan TIAN ; Baocheng HU ; Jianhong XUE
Chinese Journal of Cancer Biotherapy 1994;0(01):-
Objective:To obtain phage-displayed ScFv library targeting tumor tissues and to screen for antibodies specifically binding to tumor vessels using in vivo phage display,so as to lay a foundation for diagnosis and treatment of cancer.Methods:The membrane proteins were extracted from the specimens of esophageal carcinoma,stomach carcinoma,brain cancer,lung cancer,and spinal cord tumor.The recombinant phage-antibody system was used to construct a single-chain Fv fragment(ScFv)cDNA library from the total RNA of the BALB/c mice immunized with purified membrane protein.The specific primers of VH and VL were used to amplify the cDNA of VH and VL,respectively,which were then assembled into ScFv gene with a specially constructed linker DNA.The ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated samples were transformed into competent E.coli TG1.The transformed cells were infected with M13KO7 helper phage to yield recombinant phage.Using the animal model of human cervical carcinoma(HeLa cells),sepecific phage-ScFvs were selected by phage displaying and panning in vivo.After four rounds,24 phage-ScFvs,which were identified by PCR,were analyzed immunohistochemically.The ScFvs expressed in the tumor tissue slices and negative in control kidney tissue slices were sequenced.Results:Tomors-bearing animal models were established with 7 different kinds of carcinoma cell lines in BALB/c nude mice.It was found that inoculation with HeLa cells resulted in most satisfactory tumorigenesis in nude mice.A ScFv library of 1.6?106 was obtained and a tumor vessel specific phage-ScFv named ScFvH1(VH-linker-VL)was selected from the library.Conclusion:A tumor targeting ScFv library has been successfully constructed and a tumor vessel-specifrc antibody has been identified from the library,which provides a new way for the early diagnosis and therapy of cancer.
3.Tumor vessel-specific antibody ScFvH1: Targeting efficiency and anti-tumor ability
Xi QIN ; Hanghang MA ; Jianhong XUE ; Baocheng HU
Chinese Journal of Cancer Biotherapy 1995;0(02):-
Objective: To investigate the targeting and anti-tumor ability of the tumor vessel-specific antibody ScFvH1 selected from phage-ScFv library, and to discuss the application of the antibody in clinical diagnosis and therapy of cancer. Methods: The ScFvH1 gene was inserted into pET-28a(+)/EGFP vector containing green fluorescent protein(GFP) gene and pTIG-Trx vector containing thioredoxin gene; the products were then expressed in E.coli and purified by using Ni-NTA. Tumor-bearing mice model was established by subcutanuous injection of cervical cancer cell line HeLa. The mice were injected with purified ScFv-EGFP fusion protein through vena caudalis and the GFP signals were observed by fluorescent microscope to evaluate the targeting ability of the antibody. Meanwhile, the mice model also received intratumoral injection of purified ScFv-EGFP fusion protein to evaluate the anti-tumor effect of the antibody. Results: Soluble ScFvH1 gene and ScFvH1-EGFP protein were successfully expressed in E.coli; a single band was showed in SDS-PAGE after the purification by Ni-NTA. We found that ScFvH1-EGFP fusion protein was enriched to tumor tissues, but there was only weak fluorescent signal when EGFP protein was injected. No EGFP signal was observed in the lung of tumor-bearing mice. Tumor inhibition experiment showed that the tumor growth in the antibody treatment group was similar to that of the PBS control group. Conclusion: The tumor vessel-specific antibody ScFvH1 selected from phage-ScFv library can specifically target tumor vessels, but it has no obvious inhibitory effect on tumor growth. Our findings pave a way for antibody in cancer diagnosis and treatment.
4.Construction of the Antibody Library and Selection of the Antibodies Specifically Binding to the Prostate Carcinoma Cells
Yuan TIAN ; Xi QIN ; Baocheng HU ; Cuifen HUANG
Chinese Journal of Cancer Biotherapy 1994;0(01):-
Objective: To obtain phage-displayed ScFv library directly against prostate carcinoma cells, and select antibodies binding to prostate carcinoma cells specifically, so as to lay a foundation for developing diagnostic agents and clinical therapies of prostate carcinoma. Methods: Balb/c mice were immunized i. p . with purified membrane protein mixture of prostate carcinoma cells PC3, DU145. mRNA was isolated from the spleens of immunized mice, heavy and light chain genes ( VH and VL) of antibody were amplified separately by RT-PCR and assembled into ScFv gene with a specially constructed linker DNA. , the ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13K07 helper phage to yield recombinant phage. After five rounds of panning with PC3 cells, the positive clones were selected with the ELISA from the enriched phages. Results: A ScFv library of 3. 5 ? 106 was obtained and one phage-ScFv which can bind specifically PC3 cells was found. Conclusions: A prostate carcinoma specific antibody was identified , which paves a way for study of prostate carcinoma.
5.Experimental research into the effects of Yumian Decoction anti-asthma formula(DYA) on the plasma cAMP、cGMP and PAF contents in asthmatic guinea pigs
Chu LI ; Zijiang LONG ; Zhen WANG ; Baocheng HU ;
Chinese Traditional Patent Medicine 1992;0(05):-
Objective: To investigate the anti asthmatic effects and its mechanism of Yumian Anti asthma Formula (DYA) on asthmatic guinea pigs. Methods: Observe the change of the latent period of asthma inducing of normal guinea pigs and that of them after DYA preventing. Determine the plasma cAMP and cGMP contents. Results: The latent period of asthma inducing was obviously prolonged in guinea pigs prevented by DYA when compared with that of normal guinea pigs and guinea pigs before given DYA ( P
6.Effects of Gubi Decoction on serum related inflammatory factors and PI3K/Akt/mTOR signaling pathway in osteoarthritis model rats
Liqing ZHONG ; Shaobo DING ; Runkai HU ; Weichao HAN ; Shufen HE ; Baocheng XIE
International Journal of Traditional Chinese Medicine 2021;43(4):357-362
Objective:To explore the effect of Gubi Decoction on serum related inflammatory factors and PI3K/Akt/mTOR signaling pathway in osteoarthritis model rats. Methods:Seventy SPF rats were randomly divided into the blank group, sham operation group, Glucosamine sulfate group, and the low, medium and high dose Gubi Decoction groups. Except the blank group and sham operation group, knee osteoarthritis animal models were prepared by the modified Hulth method in each group. On the 28th day after successful model preparation, the high, medium and low dose Gubi Decoction groups were given Gubi Decoction 24, 12 and 6 g/kg by gavage respectively; glucosamine sulfate group was given glucosamine sulfate tablet suspension 3 g/L by gavage, once a day for 28 days. The levels of TNF-α and IL-1β in serum were detected by ELISA. The gene expressions of PI3K, Akt and mTOR in cartilage tissue were detected by Real-PCR. The protein expressions of PI3K, Akt, p-PI3K, p-Akt and mTOR were detected by Western blot. Results:Compared with the model group, the knee joint diameter[(11.17 ± 1.81) mm, (11.60 ± 1.38) mm, (10.80 ± 1.17) mm vs. (12.57 ± 0.98) mm] of the rats in the glucosamine sulfate group and the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05). The content of TNF-α [(111.43 ± 21.98) ng/L, (53.42 ± 13.25) ng/L vs. (157.89 ± 23.60) ng/L], IL-1β [(67.50 ± 18.44) ng/L, (48.22 ± 9.63) ng/L vs. (96.11 ± 14.85) ng/L] in the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05), and the expression of PI3K (1.87 ± 0.17, 1.24 ± 0.49 vs. 2.19 ± 0.47), Akt (1.50 ± 0.51, 1.10 ± 0.32 vs. 2.68 ± 0.63), and mTOR (1.32 ± 0.54, 1.10 ± 0.33 vs. 2.94 ± 0.55) mRNA in the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05). The expression of PI3K, Akt, p-PI3K, p-Akt in the low, medium and high dose Gubi Decoction groups significantly decreased ( P<0.05), and the expression of mTOR in the medium dose Gubi Decoction group significantly decreased ( P<0.05). Conclusion:Gubi Decoction can significantly reduce the level of inflammatory factors in the serum of osteoarthritis model rats, and its anti-inflammatory and analgesic effects may be related to PI3K/Akt/mTOR signaling pathway.
7.A method for rapid analysis of polysorbate 80 components
Ying ZHOU ; Xun ZHAO ; Nan HU ; Baocheng WANG ; Yaozuo YUAN
Journal of China Pharmaceutical University 2022;53(2):192-199
Due to the complex components of polysorbate 80, analysis is time-consuming and labor-intensive, so there is an urgent need to find a method for rapid analysis of polysorbate 80 components.In this study, 10 batches of samples collected from 3 domestic and foreign enterprises were analyzed by UHPLC-HRMS, with the being further results were analyzed by the ExcipientProfiler software and supplemented by the extended database.The results showed that the ExcipientProfiler software could quickly identify the [M+Na]+ peak in the mass spectrogram, and obtain the information of component distribution, the numbers of components and the degree of polymerization of the sample.Meanwhile, the numbers of components obtained by the ExcipientProfiler software could be used to distinguish the injection grade samples from the ordinary grade samples by systematic clustering analysis.In addition, it was found through further supplement that the sample contained other fatty acid ester components by manually searching the relevant extended database.The polyoxyethylene sorbitan tetraoleate components were found in the sample according to the analysis of mass spectrum data.Therefore, although this method is fast and simple, it is necessary to add polyoxyethylene sorbitan tetraoleate components and other fatty acid ester components to further supplement the information in the ExcipientProfiler software, so that it can be better used for the analysis of polysorbate 80.
8.Anatomical and clinical study of a novel anterial cubital approach for ulnar coronoid fractures
Baocheng ZHAO ; Tianxiang YUAN ; Xinlong MA ; Jinli ZHANG ; Baotong MA ; Jianxiong MA ; Wu YUAN ; Fangke HU ; Xiang SUN
Chinese Journal of Orthopaedics 2015;(8):859-864
Objective To investigate the feasibility of a noval anterior cubital approach for the coronoid via flexor?prona?tor teres interval and assess the clinical result. Methods Five formalin?fixed adult cadaver elbows were used. Through a single universal anteromedial longitudinal skin incision, the coronoid tip was exposed via pronator and flexor carpiradialis interval, and coronoid anteromedial facet and base via palm longus and flexor carpi ulnaris interval. The distances from the entry point to the muscles or branching point of the nerves to the line passing through medial and lateral epicondyles, as well as the length were mea?sured with regard to the motor nerve branches arising from median nerve to pronator teres, flexor carpiradialis, palm longus and flexor digiti superficialis, as well as the most proximal two motor branches to flexor carpi ulnaris arising from ulnar nerve. From September 2013 to August 2014, 4 male patients with ulnar coronoid fracture were treated operatively through the above anterior cubital approach in our hospital. They were all left side involved, with an average age of 32 years (range, 16-42 years). According to O’Driscoll classification, there were two cases of type Ib and two cases IIb respectively. They were all treated by open reduction and internal fixation through flexor?pronator teres interval. Results At cubital fossa, there were 2-3 branches to the pronator teres mostly, 1 branch to flexor carpiradialis and palm longus arising from median nerve. The branch to the flexor digiti superficia?lis usually was long and thick, and divided into 2-5 short twigs near muscle. The branch to palm longus had the same trunk with that to flexor digiti superficialis. The branch to flexor digiti superficialis was the most proximal among those passed through the in?terval of pronator teres and flexor carpiradialis, and its entry point to the muscle had an averaged distance of 37.22 mm to the line passing through medial and lateral humeral epicondyles. It was optimal to expose coronoid tip through the interval of pronator teres and flexor carpiradialis. It was safe to expose coronoid proximal to the branch to flexor digiti superficialis. While, it was better to expose the anteromedial facet and base of corocoid through the interval of palm longus and flexor carpiulnaris via median and ul?nar nerve interface. All of the four patients were followed up for an average period of 9 months. They all achieved bone union from 6 weeks to 3months (mean, 9 weeks). All the patients obtained excellent result according to the modified An&Morrey elbow per?formance index with scores from 94 to 100. Conclusion The novel anteromedial cubital approach via flexor?pronator teres is opti?mal for exposure of coronoid.
9.Clinical significance of the preoperative neutrophil lymphocyte ratio in the evaluation of the prognosis of laryngeal carcinoma
Guofeng ZHAO ; Yanhong HU ; Ruli LIU ; Feng SHI ; Haipeng LI ; Donghai WANG ; Baocheng WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2016;51(2):112-116
Objective To explore the effect of preoperative neutrophil lymphocyte ratio (NLR) on the prognosis of patients with laryngeal carcinoma.Methods Clinical data of 202 patients with laryngeal carcinoma treated from January 2004 to October 2009 were retrospective analyzed to determine the optimal critical value of NLR.To study whether NLR is an independent factor affecting the recurrence and 5-year survival rate of patients with laryngeal cancer after surgery,single factor and multivariate analyses were performed.The factors included age,gender,T stage,pathological differentiation,lymph node metastasis,primary tumor site and NLR value.The relationship between NLR and cervical lymph node metastasis was analyzed.Results The optimal critical value of NLR was 2.85,by which cases were divided into high NLR group (NLR≥2.85) and low NLR group (NLR < 2.85).Single factor and multivariate analyses indicated that T staging,lymph node metastasis,primary tumor location,and NLR were the independent factors affecting the recurrence of laryngeal carcinoma.T stage and lymph node metastasis were the independent factors affecting 5-year survival rate of laryngeal carcinoma.The increase of NLR value increased the rate of cervical lymph node metastasis.Conclusion Preoperative NLR level influences the recurrence and cervical lymph node metastasis of laryngeal carcinoma and can be considered a prognosis factor of laryngeal cancer.
10.Construction of a model based on multipoint full-layer puncture biopsy for predicting pathological complete response after neoadjuvant therapy for locally advanced rectal cancer
Ying JIN ; Zhiwei ZHAI ; Liting SUN ; Pingdian XIA ; Hang HU ; Chongqiang JIANG ; Baocheng ZHAO ; Hao QU ; Qun QIAN ; Yong DAI ; Hongwei YAO ; Zhenjun WANG ; Jiagang HAN
Chinese Journal of Gastrointestinal Surgery 2024;27(4):403-411
Objective:To investigate the value of transanal multipoint full-layer puncture biopsy (TMFP) in predicting pathological complete response (pCR) after neoadjuvant radiotherapy and chemotherapy (nCRT) in patients with locally advanced rectal cancer (LARC) and to establish a predictive model for providing clinical guidance regarding the treatment of LARC.Methods:In this multicenter, prospective, cohort study, we collected data on 110 LARC patients from four hospitals between April 2020 and March 2023: Beijing Chaoyang Hospital of Capital Medical University (50 patients), Beijing Friendship Hospital of Capital Medical University (41 patients), Qilu Hospital of Shandong University (16 patients), and Zhongnan Hospital of Wuhan University (three patients). The patients had all received TMFP after completing standard nCRT. The variables studied included (1) clinicopathological characteristics; (2) clinical complete remission (cCR) and efficacy of TMFP in determining pCR after NCRT in LARC patients; and (3) hospital attended, sex, age, clinical T- and N-stages, distance between the lower margin of the tumor and the anal verge, baseline and post-radiotherapy serum carcinoembryonic antigen (CEA) and carbohydrate antigen (CA)19-9 concentrations, chemotherapy regimen, use of immunosuppressants with or without radiotherapy, radiation therapy dosage, interval between surgery and radiotherapy, surgical procedure, clinical T/N stage after radiotherapy, cCR, pathological results of TMFP, puncture method (endoscopic or percutaneous), and number and timing of punctures. Single-factor and multifactorial logistic regression analysis were used to determine the factors affecting pCR after NCRT in LARC patients. A prediction model was constructed based on the results of multivariat analysis and the performance of this model evaluated by analyzing subject work characteristics (ROC), calibration, and clinical decision-making (DCA) curves. pCR was defined as complete absence of tumor cells on microscopic examination of the surgical specimens of rectal cancer (including lymph node dissection) after NCRT, that is, ypT0+N0. cCR was defined according to the Chinese Neoadjuvant Rectal Cancer Waiting Watch Database Study Collaborative Group criteria after treatment, which specify an absence of ulceration and nodules on endoscopy; negative rectal palpation; no tumor signals on rectal MRI T2 and DWI sequences; normal serum CEA concentrations, and no evidence of recurrence on pelvic computed tomography/magnetic resonance imaging.Results:Of the 110 patients, 45 (40.9%) achieved pCR after nCRT, which was combined with immune checkpoint inhibitors in 34 (30.9%). cCR was diagnosed before puncture in 38 (34.5%) patients, 43 (39.1%) of the punctures being endoscopic. There were no complications of puncture such as enterocutaneous fistulae, vaginal injury, prostatic injury, or presacral bleeding . Only one (2.3%) patient had a small amount of blood in the stools, which was relieved by anal pressure. cCR had a sensitivity of 57.8% (26/45) for determining pCR, specificity of 81.5% (53/65), accuracy of 71.8% (79/110), positive predictive value 68.4% (26/38), and negative predictive value of 73.6% (53/72). In contrast, the sensitivity of TMFP pathology in determining pCR was 100% (45/45), specificity 66.2% (43/65), accuracy 80.0% (88/110), positive predictive value 67.2% (45/67), and negative predictive value 100.0% (43/43). In this study, the sensitivity of TMFP for pCR (100.0% vs. 57.8%, χ 2=24.09, P<0.001) was significantly higher than that for cCR. However, the accuracy of pCR did not differ significantly (80.0% vs. 71.8%, χ 2=2.01, P=0.156). Univariate and multivariate logistic regression analyses showed that a ≥4 cm distance between the lower edge of the tumor and the anal verge (OR=7.84, 95%CI: 1.48-41.45, P=0.015), non-cCR (OR=4.81, 95%CI: 1.39-16.69, P=0.013), and pathological diagnosis by TMFP (OR=114.29, the 95%CI: 11.07-1180.28, P<0.001) were risk factors for pCR after NCRT in LARC patients. Additionally, endoscopic puncture (OR=0.02, 95%CI: 0.05-0.77, P=0.020) was a protective factor for pCR after NCRT in LARC patients. The area under the ROC curve of the established prediction model was 0.934 (95%CI: 0.892-0.977), suggesting that the model has good discrimination. The calibration curve was relatively close to the ideal 45° reference line, indicating that the predicted values of the model were in good agreement with the actual values. A decision-making curve showed that the model had a good net clinical benefit. Conclusion:Our predictive model, which incorporates TMFP, has considerable accuracy in predicting pCR after nCRT in patients with locally advanced rectal cancer. This may provide a basis for more precisely selecting individualized therapy.