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Biomedical subjects

Yong Chen

Publications and source records attributed to Yong Chen.

At least 73 records · Page 4Linked to original sources

Thermostabilization of ovotransferrin by anions for pasteurization of liquid egg white.

The effects of anions on the thermostability of ovotransferrin (oTf) were investigated. The temperature, T(m), causing aggregation of oTf was measured in the presence or absence of anions, and the denaturation temperature, T(m)(DSC), was also determined by differential scanning calorimetry (DSC) in the presence of the citrate anion. We found that some anions (phosphate, sulfate and citrate) raised temperature T(m) of oTf by about 5-7 degrees C. However, neither sodium chloride nor sodium bicarbonate raised T(m) by that much. Temperature T(m) was increased by increasing the concentration of the citrate anion, and was in good agreement with denaturation temperature T(m)(DSC), suggesting that denaturation of the oTf molecules resulted in aggregation of oTf. We also demonstrated that the anions, especially sulfate, repressed the heat-aggregation of liquid egg white. The Van't Hoff plot from the T(m) and DeltaH(d) values revealed that two anion-binding sites were concerned with heat stabilization. These binding sites may have been concerned with sulfate binding (not bicarbonate binding) that is found in the crystal structure of apo-form of oTf, since the bicarbonate anion did not raise T(m).

Animals↗

Effects of [Nphe1]nociceptin(1-13)NH2, [Phe1(CH2-NH)Gly2]nociceptin(1- 13)NH2, and nocistatin on nociceptin inhibited constrictions of guinea pig isolated bronchus.

Electric field stimulation (EFS) causes excitatory non adrenergic-non cholinergic (eNANC) and cholinergic constrictions in the guinea pig isolated bronchus, the activation of eNANC and cholinergic nerves respectively. We investigated the effects of [Nphe1]nociceptin(1-13)NH2 ([Nphe1]NC(1-13)NH2), [Phe1(CH2-NH)Gly2]nociceptin(1- 13)NH2 ([F/G] NC(1-13)NH2), and nocistatin (NST) on nociceptin (NC) inhibited constrictions in isolated bronchus of guinea pig. The results show that NC (1 micromol/L) inhibited EFS-induced eNANC and cholinergic constrictions compared with the control, in which nociceptin was not applied. After pretreatment with [Nphe1]NC(1-13)NH2, [F/G]NC(1-13)NH2, or NST, the inhibitions of NC were antagonized by [Nphe1]NC(1-13)NH2 and [F/G]NC(1-13)NH2 but not NST. However, [Nphe1]NC(1-13)NH2, [F/G]NC(1-13)NH2, and NST did not affect the inhibitions induced by morphine. Furthermore, [Nphe1]NC(1-13)NH2, [F/G]NC(1-13)NH2 and NST did not cause any appreciable effects on EFS-induced eNANC and cholinergic constrictions in guinea pig bronchi. The results demonstrate that [Nphe1]NC(1-13)NH2 and [F/G]NC(1- 13)NH2 but not NST act as selective antagonists of the NC receptor and the effects of NC on EFS-induced constrictions of guinea pig isolated bronchus.

Animals↗

Differentiation of human embryonic stem cells to hepatocytes using deleted variant of HGF and poly-amino-urethane-coated nonwoven polytetrafluoroethylene fabric.

Human embryonic stem (hES) cells have recently been studied as an attractive source for the development of a bioartificial liver (BAL). Here we evaluate the differentiation capacity of hES cells into hepatocytes. hES cells were subjected to suspension culture for 5 days, and then cultured onto poly-amino-urethane (PAU)-coated, nonwoven polytetrafluoroethylene (PTFE) fabric in the presence of fibroblast growth factor-2 (bFGF) (100 ng/ml) for 3 days, then with deleted variant of hepatocyte growth factor (dHGF) (100 ng/ml) and 1% dimethyl sulfoxide (DMSO) for 8 days, and finally with dexamethasone (10(-7) M) for 3 days. The hES cells showed gene expression of albumin in a time-dependent manner of the hepatic differentiation process. The resultant hES-derived hepatocytes metabolized the loaded ammonia and lidocaine at 7.8% and 23.6%, respectively. A million of such hepatocytes produced albumin and urea at 351.2 ng and urea at 7.0 microg. Scanning electron microscopy showed good attachment of the cells on the surface of the PTFE fabric and well-developed glycogen rosettes and Gap junction. In the present work we have demonstrated the efficient differentiation of hES cells to functional hepatocytes. The findings are useful to develop a BAL.

Actins↗

[Clinicopathologic features of 18 cases of cryptogenic organizing pneumonia].

OBJECTIVE: To analyze the characteristics of pathologically proved cases of cryptogenic organizing pneumonia (COP) including manifestation of CT scan, bronchoalveolar lavage (BAL) fluid cytology, pulmonary histopathological appearance. METHODS: Twenty-three consecutive patients from April 1994 to October 2005 in Peking Union Medical College Hospital with pathologic features of organizing pneumonia were enrolled in the study. RESULTS: Eighteen patients were diagnosed as COP. Percutaneous lung biopsy/transbronchial lung biopsy was performed in 10 patients, while open lung biopsy/video-assistant thorascopic biopsy was performed in 8 patients. There were 8 males and 10 females, aged (53.5 +/- 11.0) yr. Dyspnea, cough, inspiratory crackles were the most common symptoms and signs. Lung function tests showed a restrictive ventilatory defect (11/18), and impairment of carbon monoxide transfer factor (T(LCO), 18/18). Chest computed tomography scan in 18 cases mainly showed ground glass opacities (4/18), patchy areas of alveolar consolidation (13/18) with air bronchograms (3/18), and pleural effusion (4/18). BAL fluid cytological analysis showed the percentage of lymphocyte was 0.40 +/- 0.16, neutrophil 0.15 +/- 0.08, eosinophil 0.05 +/- 0.03; the CD(4)/CD(8) ratio was 0.43 +/- 0.21. All patients were treated with corticosteroids. Sixteen were alive and clinically improved or stabilized at the last follow up (8.7 +/- 6.2 months after diagnosis). Two patients died, one of respiratory failure, and another of pulmonary infection. CONCLUSIONS: The diagnosis of COP is usually suggested by clinical-radiological findings, but needs to be confirmed histopathologically. The combination of BAL fluid cytological findings and lung histological appearance obtained during a bronchoscopic procedure appears to be an effective method for diagnosing this disease.

Adult↗

Human membrane protein Tim-3 facilitates hepatitis A virus entry into target cells.

In this study, a cellular surface membrane protein of immunoglobulin (Ig) superfamily (IgSF) was identified from a human dendritic cell (DC) cDNA library by large-scale random sequencing, which is identical to previously reported Tim-3 (T-cell Ig- and mucin-domain-containing molecule 3). Recent data have suggested the association of the 281-residue mouse Tim-3 molecule with Th1-related T cell responses and disease in mice. Human Tim-3 is a 301-residue type I membrane protein whose extracellular region contains a Cys-rich Ig-like domain and a mucin domain, the characteristics of Tim proteins. It shows significant homology to human hepatitis A virus (HAV) cellular receptor-1 (HuHAVcr-1)/Tim-1. Human Tim-3 mRNA was highly expressed in monocytes or monocyte-derived cells, and the expression level decreased when DC underwent maturation and activation. There is no previous report on the biological functions of human Tim-3, especially the involvement in virus infection. We demonstrated that HeLa cells, which are refractory to HAV infection, acquired a limited susceptibility to HAV infection after stably overexpressing human Tim-3 as confirmed by Western blot analysis using anti-Tim-3 antibody, but Tim-3-Fc fusion protein had no direct HAV-binding activity. The results indicated that human Tim-3 can promote HAV entry into target cells but itself may not function as a cellular receptor of HAV.

Amino Acid Sequence↗

[HPLC-MSn analysis of trigonelline and its metabolites in rat urine].

AIM: To establish a rapid and sensitive LC-MSn method for the identification of trigonelline and its main metabolites in rat urine. METHODS: After optimizing the detection conditions of LC-MSn chromatography and mass spectrometry using trigonelline, its ionization and cleavage in ESI-MS and ESI-MSn modes were summarized, then serving as the basis for the metabolite analysis of trigonelline in rat urine. The 0-48 h urine samples of rats were collected after iv 8 mg x kg(-1) trigonelline, then, the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by LC-MSn. RESULTS: The structures of trigonelline metabolites were elucidated according to the changes of the molecular weights of the metabolites (deltaM) and their cleavage pattern in ESI-ITMSn. As a result, two phase I metabolites and the parent drug were identified existing in rat urine, and two phase II metabolites were identified. CONCLUSION: The LC-MSn method is rapid and high sensitive and specific, it is suitable for the identification of trigonelline and its metabolites in rat urine.

Alkaloids↗

[Preparation and characterization of monoclonal antibody against human LSECtin].

AIM: To prepare and characterize monoclonal antibody (mAb) against human LSECtin (liver and lymph node sinusoidal endothelial cell C-type lectin) protein. METHODS: BALB/c mice were immunized with prokaryotically expressed human LSECtin protein. The splenocytes from the immunized mice were fused with murine myeloma cells (Sp2/0) and then the mAb-positive hybridoma cells were screened by indirect ELISA. Reaction of mAb to LSECtin antigen was characterized by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM. RESULTS: Eight hybridoma cells secreting mAbs were established. The isotypes of the mAbs were IgG. Ascites titers were between 1:10(6) - 1:10(7). All the mAbs recognized human LSECtin protein on LSECtin-transfected 3T3 cells and six of the mAbs specifically recognized liver sinusoidal endothelial cells. CONCLUSION: Eight anti-LSECtin mAbs have been obtained. The characterization of the mAbs indicate that they show fine specificity by Western blot, indirect immunofluorescent staining, immunohistochemical staining and FCM, which can provide a powerful reagent for the functional study of LSECtin.

Animals↗

[HPLC-ESI/MS analysis of stachydrine and its metabolites in rat urine].

AIM: To identify the main metabolites of stachydrine in rat. METHODS: The ionization, cleavage and chromatographic characteristics of stachydrine were studied by using high-performance liquid chromatography-electrospray ionization ion trap tandem mass spectrometry (HPLC-ESI/MS) for the first time. These characteristics of stachydrine were used as the basis for the analyses of metabolites in rat urine. The 0 - 24 h urine samples of rats after ig 25 mg x kg(-1) stachydrine were collected and purified by using C10 solid-phase extraction cartridge, and then analyzed by HPLC-ESI/MS to identify stachydrine and its metabolites. RESULTS: The parent drug (stachydrine), 6 phase I metabolites (N-demethyl, dehydrogenation, ring-oxidation) and 2 phase II metabolites (glycine conjugates of 2 ring-oxidation products) were identified existing in rat urine. CONCLUSION: The presented method was proved to be sensitive, rapid, high selective and specific for the identification of stachydrine and its metabolites in rat urine.

Animals↗

Maintenance of mouse, rat, and pig pancreatic islet functions by coculture with human islet-derived fibroblasts.

Development of an efficient preculture system of islets is ideal. Toward that goal, we constructed a human pancreatic islet-derived fibroblast cell line MNNK-1 for a source as a coculture system for freshly isolated islets to maintain islet functions. Human pancreatic islet cells were nucleofected with a plasmid vector pYK-1 expressing simian virus 40 large T antigen gene (SV40T) and hygromycin resistance gene (HygroR). One of the transduced cell lines, MNNK-1, was established and served as a feeder cell in the coculture for freshly isolated mouse, rat, and pig islets. Morphology, viability, and glucose-responding insulin secretion were analyzed in the coculture system. MNNK-1 cells were morphologically spindle shaped and were negative for pancreatic endocrine markers. MNNK-1 cells were positive for alpha-smooth muscle actin and collagen type I and produced fibroblast growth factor. Coculture of the mouse, rat, and pig islets with MNNK-1 cells maintained their viability and insulin secretion with glucose responsiveness. A human pancreatic islet-derived fibroblast cell line MNNK-1 was established. MNNK-1 cells were a useful means for maintaining morphology and insulin secretion of islets in the coculture system.

Actins↗

[Analysis of anisodine and its metabolites in rat plasma by liquid chromatography-tandem mass spectrometry].

AIM: To identify anisodine and its metabolites in rat plasma after ingestion of anisodine by combining liquid chromatography and tandem mass spectrometry (LC-MS(n)). METHODS: Plasma samples from rats after a single orally administration of 20 mg anisodine were added with methanol to precipitate protein. Then, it was analyzed by LC-MS(n). Identification and structural elucidation of the metabolites were performed by comparing their changes in molecular masses, retention-times and full scan MS(n) spectra with those of the parent drug and blank plasma. RESULTS: The results revealed that the parent drug and its four metabolites (norscopine, scopine, hydroxyanisodine, N-oxide anisodine) existed in rat plasma. CONCLUSION: This method is sensitive, rapid, simple, and it is suitable for the rapid identification of drug and its metabolits.

Administration, Oral↗

[Quercetin inhibits growth and induces apoptosis of human gastric carcinoma cells].

AIM: To study the effect of quercetin on the growth and apoptosis of human gastric carcinoma cell line MGC-803. METHODS: The measurement of inhibitory rate and apoptotic index(AI) of quercetin were done by MTT assay and TUNEL assay. The positive expression rate of P53, C-myc and P16 were detected by immunocytochemical staining. RESULTS: Quercetin at concentrations ranging from 40 mumol/L to 100 mumol/L significantly inhibited the proliferation of MGC-803 cells in a dose- and time-dependent manner (P<0.01). TUNEL assay indicated that the number of apoptotic cells in quercetin-treated group was greater than that in the control group (P<0.01). Expression of P53 and C-myc protein decreased following quercetin induction in a dose-dependent manner, whereas P16 expression increased significantly compared with that of the control group (P<0.01). CONCLUSION: Quercetin can inhibit the growth and induce apoptosis of MGC-803 cells in a dose- and time-dependent manner. Its mechanisms may be relevant to the down-regulation of P53 and C-myc protein expression as well as up-regulation of P16 expression.

Apoptosis↗

[Diagnosis of recurrent or residual nasopharyngeal carcinomas in the skull base area with F-18-fluoro-deoxyglucose positron emission tomography].

OBJECTIVE: To investigate the diagnostic value of F-18-fluoro-deoxyglucose positron emission tomography (FDG-PET) for the recurrent or residual nasopharyngeal carcinomas in the skull base area. METHODS: Nine post-irradiation nasopharyngeal carcinoma patients did FDG-PET scanning, CT/MRI imaging and underwent nasopharynx and skull base-biopsy under endoscopy. The results of FDG-PET were evaluated and compared with CT/MRI studies and biopsies. RESULTS: In 9 cases of post-irradiation nasopharyngeal carcinoma, CT/MRI detected 7 recurrent cases and 2 suspected recurrent cases in occipital bone and clivus. All 9 cases had accumulated FDG in nasopharynx and cranial base. A definite diagnosis was made by biopsy, 3 cases were confirmed recurrence, and others 6 cases were proved mucous chronic inflammation and (or) osteoradionecrosis. The accuracy of FDG-PET was 33.3% (3/9), and the false positive rate was 66.7% (6/9). CONCLUSIONS: Diagnosis of recurrent or residual nasopharyngeal carcinomas in the skull base area with FDG-PET had high false-positive rate, final diagnosis must depend on histopathologic examination under endoscopy.

Adult↗

Treatment of chronic osteomyelitis with one-stage allograft.

OBJECTIVE: To avoid disadvantages of two-stage cancellus bone autograft, we investigated the feasibility of one-stage allograft for reconstructing the bone defect resulting from debridement of chronic osteomyelitis in limbs. METHODS: Between Feb. 1999 and Apr. 2004, 35 cases of chronic osteomyelitis (8 cases of nonunion) underwent one-stage allograft after debridement in our hospital. RESULTS: Thirty-five cases were followed up for an average period of 28 months (range, 13 to 55 months), in which 32 cases (91.43%) were found no infection, and 3 cases (8.57%) were confirmed recurrence of infection. Four out of 8 cases of bone nonunion healed in 9.5 months on average (range, 3 to 12 months), and another case also acquired union after redebridement and autograft of ilium due to infection recurrence 35 days after surgery. Renonunion occurred in 3 cases, 2 out of whom healed after secondary operation with autograft. One case of renonunion and 2 cases of infection recurrence refused further treatment. CONCLUSIONS: A high rate of infection arrest can be attained when one-stage allograft is used to reconstruct the bone defect of chronic osteomyelitis after debridement in limbs. Therefore, chronic osteomyelitis should not be regarded as a contraindication to one-stage allogeneic bone grafting. Renonuion, however, achieves a relatively high rate, especially in cases of segmental bone defect.

Adolescent↗

[Selective three-field lymphadenectomy for thoracic esophageal squamous carcinoma].

OBJECTIVE: To investigate the lymph node metastasis and the rational lymphadenectomy in thoracic esophageal carcinoma. METHODS: Eighty-seven patients with thoracic esophageal squamous carcinoma received esophagectomy plus two-field or three-field lymphadenectomy based on cervical ultrasonography. RESULTS: Thirty-five patients (40.2% ) with enlarged cervical nodes revealed by cervical ultrasonography received cervical lymphadenectomy. The proportion of cervical lymphadenectomy was 66.7% (16/24) in upper thoracic esophageal carcinomas, significantly higher than 30.2% (19/63) in middle and lower esophageal carcinomas (P=0.002). Regional and cervical lymph node metastasis were found in 48(55.2% ) and 17(19.5% ) patients respectively. The regional lymph node metastatic rates were 37.5% (9/24), 62.3% (33/53) and 60.0% (6/10) respectively in the patients with upper, middle, and lower thoracic esophageal carcinoma. The cervical lymph node metastatic rates in the patients with or without regional lymph node metastasis were 31.3% (15/48) and 5.1% (2/39) respectively(P=0.002). The rates of upper, mid, lower mediastinal and upper abdominal lymph node metastasis were 25.3%, 23.0%, 5.7%, and 24.1% respectively. Cervical lymph node metastasis was significantly correlated with upper and mid mediastinal metastasis (both P< 0.01), but not with lower mediastinal and upper abdominal lymph node metastasis. The overall postoperative morbidity rate was significantly higher in three field lymphadenectomy group than that in two field group(60.0% vs. 34.6%, P=0.020). CONCLUSION: Selective 3-field lymphadenectomy based on cervical ultrasonography should be performed in thoracic esophageal carcinoma, especially with upper and mid mediastinal lymph node metastasis.

Adult↗

[Determination of cyclovirobuxine-D in huangyangning tablets by RP-HPLC with gradient elution].

OBJECTIVE: To establish a new method for the determination of cyclovirobuxine-D in huangyangning tablets. METHOD: Agilent Zorbax Extend C18 (4.6 mm x 250 mm, 5 microm) column with temperature 40 degrees C was used. The mobile phase was 0.3% diethylamine methanol solution and 0.3% diethylamine water solution, with a linear gradient of 0.3% diethylamine methanol solution from 78% to 95% in 9 minutes, and then maintained for 6 minutes. The flow rate was 0.8 mL x min(-1). Evaporative light scattering detector was used. RESULT: The calibration curve was linear at a range of 0.57-11.44 microg for the cyclovirobuxine D (r = 0.9996). The average recovery was 97.2% and RSD was 1.1% (n = 9). CONCLUSION: This method is rapid, simple, and reproducible, and can be used as a quality control method for this preparation.

Buxus↗

[Effects of neonatal recurrent seizures on gamma-aminobutyric acid B1 receptor expression in the rat brain].

OBJECTIVE: This study investigated the effects of flurothyl-induced neonatal recurrent seizures on gamma-aminobutyric acid B1 receptor (GABAB1R) expression in neonatal and adult rat brain, and explored the possible relationship between the alterations of GABAB1R in mature brain and the changes of spatial memory and seizure susceptibility in adult rats. METHODS: Forty-eight postnatal day (P) 7 Sprague-Dawley rats were randomly assigned into two groups: Control and Seizure group (n=24 each). Seizures were induced by inhalant flurothyl daily for six consecutive days in rat pups from the Seizure group. Twelve rats selected randomly in each group were sacrificed on the 7th day after the last seizure for detecting the expressions of GABAB1R mRNA and protein in cerebral cortex and hippocampus by reverse transcription-polymerase chain reaction (RT-PCR) and immuno-histochemistry method. The spatial memory was tested by using the Morris water maze task during P61 to P64 and the seizure threshold was measured at P75 following intraperitoneal injection of pentylenetetrazol ( PTZ ) in the remaining rats. The rats were then sacrificed for detecting the expressions of GABAB1R mRNA and protein in cerebral cortex and hippocampus. RESULTS: The expressions of GABAB1R mRNA and protein in the cerebral cortex on the 7th day after the last seizure and at P75 decreased significantly in the Seizure group when compared with the Control group (P < 0.05). The GABAB1R protein expression in the dentate gyrus on the 7th day after the last seizure in the Seizure group was significantly lower than that in the Control group (P < 0.05), but the GABAB1R mRNA expression in the hippocampus was not different from that in the Control group. There were no significant differences in the expressions of GABAB1R mRNA and protein in the hippocampus between the two groups at P75. The escape latencies in water maze of the rats in the Seizure group at P64 were significantly longer than those in the Control group (98,533.8 +/- 27,205.4 ms vs 46,723.3 +/- 40,666.5 ms; P <0.05). There were no differences in the seizure threshold between the two groups. CONCLUSIONS: The expressions of GABAB1R mRNA and protein in the cerebral cortex and hippocampus of neonatal rats with recurrent seizures decreased significantly, suggesting the changes of GABAB1R may be related to acute brain injury following neonatal recurrent seizures and the memory deficit in adult rats caused by neonatal recurrent seizures.

Animals↗

[Liquid chromatography-tandem electrospray ionization ion trap mass spectrometric assay for the metabolites of jatrorrhizine in rat urine].

AIM: To identify the main metabolites of jatrorrhizine in rat urine. METHODS: The rat urine samples were collected 0 - 72 h after ig 12 mg x kg(-1) jatrorrhizine, then the samples were purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by combining liquid chromatography and tandem electrospray ionization ion trap mass spectrometry (LC-ESI/ITMS(n)). Identification and structural elucidation of the metabolites were performed by comparing the changes in molecular masses, retention-times and full scan MS(n) spectra with those of the parent drug. RESULTS: At least seven phase I metabolites (such as de-methyl, de-hydrogen and hydroxyl metabolites) and eleven phase II metabolites (such as glucuronide conjugates and methyl-conjugates) were identified in rat urine. CONCLUSION: The developed LC-ESI/ITMS(n) method is not only simple and rapid but also sensitive and specific for the identification of metabolites of jatrorrhizine in rat urine.

Animals↗

[Optimizing process of four-effect countercurrent extraction of danshensu from Salvia miltiorrhiza].

OBJECTIVE: To investigate the process factors and characteristic of four-effect countercurrent extracting process of Danshensu from Salvia miltiorrhiza. Optimizing the parameters by using regression analysis and principal components analysis (PCA). METHOD: Applying the orthogonal design method to optimize the extracting process. The factors of extraction time, solvent volume, extraction temperature and the degree of grinding were considered. And also, the new process was compared with the refluence process. RESULT: The optimal condition of four-effect countercurrent extraction of Danshensu from S. miltiorrhiza was 1 h extraction time, solvent to sample ratio of 12 mL x g(-1), 80 degrees C , 20 - 40 mesh of particle size. Compared with the refluence process, the countercurrent extracting process has the advantages of higher recovery, less solvent consumption and shorter extraction time. CONCLUSION: The four-effect countercurrent extraction process has great advantages and is good for Chinese medicinal materials.

Lactates↗