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Biomedical subjects

Jian Huang

Publications and source records attributed to Jian Huang.

At least 91 records · Page 5Linked to original sources

Our search for the porcine mother rotor.

A single stationary mother rotor, located in the fastest activating region and giving rise to activation fronts that propagate throughout the remainder of the myocardium, has been hypothesized to be responsible for the maintenance of ventricular fibrillation (VF). Others have reported a mother rotor in guinea pigs and rabbits. We wanted to see if a mother rotor exists in a larger heart, that is, pigs. Epicardial mapping studies have demonstrated that VF wavefronts in pigs tend to propagate from the posterior basal LV to the anterior LV and on to the anterior RV, raising the possibility of a mother rotor in the posterior LV. However, no sustained reentry consistent with a mother rotor was found on the posterior LV epicardium, even though an intramural mapping study showed that the fastest activating transmural layer was near the epicardium. Many wavefronts in the posterior LV entered the mapped region from the posterior boundary of the mapping array, adjacent to the posterior descending coronary artery, raising the possibility that a mother rotor is located in the right ventricle or septum. Since a previous study has shown that the RV activates more slowly than the LV during VF, the more likely site for a mother rotor was the septum. However, we then performed a study in which we recorded from the right side of the septum and found that reentry was uncommon there also and that the activation rate was slower than the posterobasal LV. Many of the VF wavefronts in the septum passed from the posterior septum toward the anterior septum. This fact coupled with the fact that many wavefronts passed from the posterior LV free wall toward the anterior LV free wall point to the region where the posterior free wall intersects with the septum, the region where the posterior papillary muscle is located, as the possible site of a mother rotor. Indeed, a recent abstract by others reports that, after propranolol, a stable reentrant circuit is present on the endocardium at the insertion of the posterior papillary muscle into the LV free wall in pigs.

Animals↗

The Epstein-Barr virus replication protein BBLF2/3 provides an origin-tethering function through interaction with the zinc finger DNA binding protein ZBRK1 and the KAP-1 corepressor.

Herpesviruses encode a set of core proteins essential for lytic replication of their genomes. Three of these proteins form a tripartite helix-primase complex that, in the case of Epstein-Barr virus (EBV), consists of the helicase BBLF4, the primase BSLF1, and the linker protein BBLF2/3. BBLF2/3 and its homologs in the other herpesviruses remain relatively poorly characterized. To better understand the contribution to replication made by BBLF2/3, a yeast two-hybrid screen was performed with BBLF2/3 as the bait protein. This screen identified as interactors a number of cell replication-related proteins such as DNA polymerase beta and subunits of DNA polymerase delta along with the EBV-encoded DNase BGLF5. The screen also identified the DNA binding zinc finger protein ZBRK1 and the ZBRK1 corepressor KAP-1 as BBLF2/3 interactors. Interaction between BBLF2/3 and ZBRK1 and KAP-1 was confirmed in coimmunoprecipitation assays. A binding site for ZBRK1 in the EBV oriLyt enhancer was identified by electrophoretic mobility shift assay. ZBRK1, KAP-1, and the ZBRK1 binding protein BRCA1 were shown by indirect immunofluorescence to be present in replication compartments in lytically induced D98-HR1 cells, and additionally, chromatin immunoprecipitation assays determined that these proteins associated with oriLyt DNA. Replication of an oriLyt plasmid and a variant oriLyt (DeltaZBRK1) plasmid was examined in lytically induced D98-HR1 cells. Exogenous ZBRK1, KAP-1, or BRCA1 increased the efficiency of oriLyt replication, while deletion of the ZBRK1 binding site impaired replication. These experiments identify ZBRK1 as another cell protein that, through BBLF2/3, provides a tethering point on oriLyt for the EBV replication complex. The data also suggest that BBLF2/3 may serve as a contact interface for cell proteins involved in replication of EBV oriLyt.

Animals↗

Regulation of expression of the Epstein-Barr virus BamHI-A rightward transcripts.

The Epstein-Barr virus (EBV) BamHI-A rightward transcripts, or BARTs, are a family of mRNAs expressed in all EBV latency programs, including EBV-infected B cells in healthy carriers. Despite their ubiquitous expression, the regulation and biological function of BARTs are still unclear. In this study, the BART 5' termini were characterized by using a procedure that selects capped, full-length mRNAs. Two TATA-less promoter regions, designated P1 and P2, were mapped. P1 had relatively high basal activity in both epithelial and B cells, whereas P2 exhibited higher activity in epithelial cells. Upon EBV infection of B cells, transcription from P1 was detected soon after infection, while expression from P2 was delayed. Promoter-reporter assays in transiently transfected cells revealed that P1 and P2 were differentially regulated. Interferon regulatory factor 7 (IRF7) and IRF5 negatively regulated P1 activity. c-Myc and C/EBP family members positively regulated P2. Regulation of P2 by C/EBPs was characterized by electrophoretic mobility shift assay, chromatin immunoprecipitation, and reporter assays. More-abundant BART expression in epithelial cells correlated with the relative expression of positive and negative regulators in these cells.

B-Lymphocytes↗

Bcl-2 up-regulation and P-p53 down-regulation account for the low sensitivity of murine L929 fibrosarcoma cells to oridonin-induced apoptosis.

Drug resistance has been a major limitation to chemotherapy. There are many mechanisms that contribute to such resistance. In our study, we subcloned oridonin-sensitive and low sensitive L929 cells and both types of cells grew at almost the same growth rate. The acquired low sensitivity to oridonin-induced apoptosis was associated with Bcl-2 up-regulation and down-regulation of p53 phosphorylation. The p38 inhibitor SB203580 decreased Bcl-2 expression in the low sensitive L929 cells and made the cells more sensitive to oridonin. Moreover, a higher dose of oridonin promoted p53 phosphorylation, increased Bax expression and subsequently induced death of low sensitive L929 cells, however, it had no effect on Bcl-2 expression. The increased Bcl-2/Bax ratio in oridonin low sensitive L929 cells did not inhibit caspase-9 or -3 activation, but suppressed the cleavage of poly (ADP-ribose) polymerase (PARP), indicating the existence of caspase-9 or -3 independent PARP activation. These results indicated that in L929 cells, there was a relationship among the low sensitivity to oridonin, down-regulation of p53 phosphorylation and Bcl-2 up-regulation.

Animals↗

Identification of differentially expressed genes contributing to radioresistance in lung cancer cells using microarray analysis.

Radiotherapy has played a key role in the control of tumor growth in many cancer patients. It is usually difficult to determine what fraction of the tumor cell population is radioresistant after a course of radiotherapy. The response of tumor cells to radiation is believed to be accompanied by complex changes in the gene expression pattern. It may be possible to use these to sensitize radioresistant tumor cells and improve radiocurability. Based on the biological effects of ionizing radiation, in the present study, we developed one oligonucleotide microarray to analyze the expression of 143 genes in cells of two lung cancer cell lines with different radiosensitivities. Compared to NCI-H446 cells, expression of 18 genes significantly increased the basal levels in the radioresistant A549 cells, in which eight genes were up-regulated and 10 genes were down-regulated. In A549 cells irradiated with 5 Gy, 22 (19 up-regulated and three down-regulated) and 26 (eight up-regulated and 18 down-regulated) differentially expressed genes were found 6 and 24 h after irradiation, respectively. In NCI-H446 cells, the expression of 17 (nine up-regulated and eight down-regulated) and 18 (six up-regulated and 12 down-regulated) genes was altered 6 and 24 h after irradiation, respectively. RT-PCR was performed, and we found that MDM2, BCL2, PKCZ and PIM2 expression levels were increased in A549 cells and decreased in NCI-H446 cells after irradiation. Genes involved in DNA repair, such as XRCC5, ERCC5, ERCC1, RAD9A, ERCC4 and the gene encoding DNA-PK, were found to be increased to a higher level in A549 cells than in NCI-H446 cells. Antisense suppression of MDM2 resulted in increased radiosensitivity of A549 cells. Taken together, these results demonstrate the possibility that a group of genes involved in DNA repair, regulation of the cell cycle, cell proliferation and apoptosis is responsible for the different radioresistance of these two lung cancer cells. This list of genes may be useful in attempts to sensitize the radioresistant lung cancer cells.

Cell Line, Tumor↗

The augmentation of TNFalpha-induced cell death in murine L929 fibrosarcoma by the pan-caspase inhibitor Z-VAD-fmk through pre-mitochondrial and MAPK-dependent pathways.

We investigated the mechanism of the pan-caspase inhibitor z-VAD-fmk's augmentation of TNFalpha-induced L929 cell death and found this mechanism differs from that of TNFalpha-induced L929 cell death. In the presence of 20 ng/ml TNFalpha, z-VAD-fmk initiated apoptosis and necrosis in the majority of L929 cells as measured by an agarose gel electrophoresis and lactate dehydrogenase(LDH)activity based assay. Mitochondrial permeability transition (MPT) inhibitor (cyclosporine A) effectively inhibited z-VAD-fmk-augmented cell death. In addition, z-VAD-fmk plus TNFalpha increased Bax expression without affecting Bcl-2 and cytochrome expression. Western-blot analysis showed that z-VAD-fmk plus TNFalpha caused persistent JNK activation and ERK inactivation. Poly(ADP-ribose) polymerase (PARP) inhibitor (DPQ) effectively reversed the cell death which was augmented by z-VAD-fmk, and z-VAD-fmk plus TNFalpha also caused PARP cleavage to an 85 KDa fragment. These results indicate that in the presence of TNFalpha, z-VAD-fmk further augments cell death which requires the mitochondrial permeability transition and the JNK activation. However, we did not detect the changes in cytochrome c expression and the participation of caspase-9 in this process, suggesting that there might exist an unknown signal pathway(s) from the mitochondria to the downstream protein PARP, which is cleaved in a caspase-independent manner.

Amino Acid Chloromethyl Ketones↗

A comparison of the signal pathways between the TNF alpha- and oridonin-induced murine L929 fibrosarcoma cell death.

Oridonin, an active component isolated from Rabdosia rubescences, has been reported to have antitumor effects. In this study, we compared the signal transduction pathways between TNFalpha-and oridonin-induced L929 cell death. Oridonin and TNFalpha initiated apoptotic morphologic changes, but DNA fragmentation was found in TNFalpha-treated L929 cells but not in oridonin-treated ones. The pan-caspase inhibitor (z-VAD-fmk), caspase-8 inhibitor (z-IETD-fmk) and caspase-3 inhibitor (z-DEVD-fmk) augmented oridonin-and TNFalpha-induced cell death. However, the caspase-9 inhibitor (z-LEHD-fmk) only increased oridonin-induced L929 cell death. Moreover, poly (ADPribose) polymerase (PARP) was cleaved in oridonin-treated L929 cells but not in the TNFalpha-treated groups, and the caspase-3 inhibitor (z-DEVD-fmk) failed to inhibit PARP cleavage. These results showed that only oridonin-induced L929 cell death required PARP degradation in a caspase-3 independent manner. In addition, oridonin increased the ratio of Bax/Bcl-2 protein expression, but TNFalpha did not. TNFalpha induced p38 and ERK activation, whereas oridonin triggered only ERK activation. We also investigated the effect of oridonin on intracellular TNFalpha expression, and found that oridonin augmented endogenous pro-TNFalpha expression and its upstream protein IkB phosphorylation. These results indicated that although oridonin promoted endogenous pro-TNFalpha expression, a great difference existed between the signal pathways through which TNFalpha-and oridonin-induced cell death.

Animals↗

Clinical evaluation of oligonucleotide microarrays for the detection of HBV mutants associated with lamivudine resistance.

OBJECTIVES: Long-term lamivudine administration in hepatitis B virus (HBV)-infected patients induces the emergence of HBV mutants with lamivudine resistance. The aim of the present study was to evaluate the clinical application of an oligonucleotide microarray in detecting HBV mutants associated with lamivudine resistance. METHODS: 947 HBV DNA-positive sera from: 388 patients receiving lamivudine treatment, 559 chronic hepatitis B patients not receiving lamivudine treatment, and 359 from HBV DNA-negative controls, were assayed for HBV mutations using the oligonucleotide microarray. Furthermore, follow-up studies were performed using 255 clinical samples from 51 patients treated with lamivudine at various periods. The results were compared with sequencing and real-time polymerase chain reaction (PCR). RESULTS: The HBV DNA polymerase Tyr-Met-Asp-Asp motif (YMDD) mutation was detected in all 388 samples containing lamivudine-resistant mutations identified by microarray. For the codons rt180, rt204 and rt207, the agreements between the microarray and sequencing data are 96.6, 98.5 and 100%, respectively. Two previously unreported mutants were also found in those samples. In the 947 samples collected from different patients, which were detected positive for HBV DNA by quantitative PCR, all but three weak-positive samples were positive by the microarray, demonstrating an agreement of 99.7%. In all the positive samples, mutations could be detected in the relevant loci of HBV DNA polymerase with lamivudine resistance. All of the 359 HBsAg-negative samples were shown to be negative for HBV DNA using the microarray method. Follow-up detection of the clinical samples from 51 patients treated with lamivudine demonstrated that the microarray method was able to detect mutations in mixed viruses that were infecting prior to sequencing. CONCLUSION: The oligonucleotide microarray can be conveniently utilized to detect mutant HBV in clinical serum samples.

Antiviral Agents↗

Anti-SARS-CoV immunoglobulin G in healthcare workers, Guangzhou, China.

To determine the prevalence of inapparent infection with severe acute respiratory syndrome (SARS) among healthcare workers, we performed a serosurvey to test for immunoglobulin (Ig) G antibodies to the SARS coronavirus (SARS-CoV) among 1,147 healthcare workers in 3 hospitals that admitted SARS patients in mid-May 2003. Among them were 90 healthcare workers with SARS. As a reference group, 709 healthcare workers who worked in 2 hospitals that never admitted any SARS patients were similarly tested. The seroprevalence rate was 88.9% (80/90) for healthcare workers with SARS and 1.4% (15/1,057) for healthcare workers who were apparently healthy. The seroprevalence in the reference group was 0.4% (3/709). These findings suggest that inapparent infection is uncommon. Low level of immunity among unaffected healthcare workers reinforces the need for adequate personal protection and other infection control measures in hospitals to prevent future epidemics.

Adult↗

[Expressions of aromatase P450 and estrogen receptor in eutopic and ectopic endometrium in endometriosis and their correlation with endometriosis].

OBJECTIVE: To detect the expression of aromatase P450 and estrogen receptor (ER) in eutopic and ectopic endometrium in endometriosis and their correlation with endometriosis. METHODS: Forty patients who had undergone operation because of endometriosis (all were stage III-IV according to the revised American Fertility Society classification) were enrolled and 83 tissue specimens from different locations of disease foci were obtained and divided into five groups according to the location: group I, eutopic endometrium of 25 cases; Group II, ovarian endometriosis tissues of 23 cases; Group III, vagina-rectum endometriosis tissues of 11 cases; Group IV, peritoneum endometriosis tissues of eight cases; and group V, uterine serosa endometriosis tissues of 16 cases. Control group consisted of normal endometrium taken from 20 fertile women who had endometrial curettage before placement of intrauterine device. The routine two-step immunohistochemical technique was used to measure the expression of aromatase P450 and ER in endometrial cells. RESULTS: Expressions of aromatase P450 and ER in group I [histochemistry score (H-score), 2.6 +/- 1.0, 3.8 +/- 0.5] were significantly higher than those in control group (both P < 0.01), group II (1.4 +/- 1.0, 1.9 +/- 1.6) (P < 0.05, P < 0.01), and group III (1.2 +/- 0.9, 1.8 +/- 1.6) (both P < 0.05). Expressions of aromatase P450 and ER had positive correlation in all groups (r = 0.577, P < 0.01). CONCLUSIONS: Aromatase P450 could increase estrogen and estrogen receptor levels in the local ectopic tissues, thus promoting the growth and implantation of endometriosis. Different types of endometriosis focus have different levels of hormone regulation.

Adult↗

[Study on inhibitory effect of matrine on cyclooxygenase-2 expression in colon cancer HT-29 cell line].

OBJECTIVE: To explore the effect of matrine on cyclooxygenase-2 (COX-2) expression in colon cancer HT-29 cell line at the level of gene and protein. METHODS: Levels of mRNA and protein expression of COX-2, and its synthesized product prostaglandin E2 (PGE2) of colon cancer HT-29 cell line were detected by RT-PCR, Western-blot, ELISA respectively before and after treatment of matrine in different concentrations. RESULTS: Matrine had inhibitory effect on the mRNA and protein expression of COX-2, and synthesis of PGE2 in colon cancer HT-29 cell line, but had no effect on COX-1. When HT-29 cell line was treated with 2.0 mg/ml of matrine, the inhibitory rate on COX-2 mRNA expression were 100% at 6 hrs and 9 hrs after treatment; the inhibitory rate on PGE2 synthesis was 63.8 % at 9 hrs after treatment; and that on COX-2 protein expression was 48% and 100% 12 hrs and 24 hrs after treatment, respectively. CONCLUSION: Matrine has selective inhibitory effect on gene transcription, protein expression and functional activity of COX-2 in HT-29 cell line, which is time-dependent and concentration-dependent within certain range of concentration and acting time.

Alkaloids↗

[The treatment of Semont maneuver for 40 patients with posterior semicircular canal benign paroxysmal positional vertigo].

OBJECTIVE: To evaluate the effectiveness of Semont maneuver for posterior semicircular canal benign paroxysmal positional vertigo (PC-BPPV). METHOD: Forty patients with PC-BPPV were treated with Semont maneuver from January to August in 2004. RESULT: After a single treatment session of Semont maneuver,the symptoms disappeared completely in 32 patients, the first success rate was 80%. Among 19 patients who had only 1 week duration of PC-BPPU, the symptoms disappeared in 16 patients after a single treatment. The symptoms disappeared in 3 patients after twice, 2 patients after 3 times and 1 patient after 4 times of Semont maneuver sessions, the total success rate was 95%. No effectiveness was found 2 patients; PC-BPPU recurred in 5 patients during follow-up and the recurrent rate was 12.5%. it was still effective for another Semont maneuver therapy session. CONCLUSION: The Semont maneuver therapy is safe and effective for PC-BPPV. Repeated Semont maneuver therapy is still effective for them.

Adult↗

[Reducing the influence non-linearity of detecting array on absorption spectrum with curve fitting].

In a micro spectral analytical system, the non-linearity response of the photoelectronic detecting array pixel will cause the deformation of the spectrum, especially in the measurement of absorption spectrum. In this paper, the factors that cause the deformation were studied and a corresponding solution was proposed to reduce such deformation. By using curve fitting, the non-linearity of the pixel can be corrected. This method was adopted to correct the absorption spectrum obtained by a micro spectral analytical system and the result showed that it is a efficient way to correct the non-linearity in the pixel of photoelectronic detecting array.

Algorithms↗

[Agrobacterium-mediated transformation of rice mature embryos and regeneration of transgenic plants with Metr gene].

The mature embryos of a Japonica rice, Guanglingxiangnuo, were used for the study on optimization of Agrobacterium-mediated transformation. Several factors affecting the transformation were investigated and a suitable transformation system was set up. For this transformation system, the HRM medium, based on the MS medium, was suitable for inducing callus from rice mature embryos. The suitable time span of initial culture in this medium was 7-8 days before co-culturing with Agrobacterium and suitable medium for selection was CC medium. Using this transformation system, the Metr gene was introduced into Guanglingxiangnuo, and many transgenic plants were obtained. Most of these transgenic rice plants were confirmed by PCR technique and basta resistance, indicating the T-DNA had been integrated into the genome of transgenic rice plants.

Arabidopsis Proteins↗

[Identification of response element gene sequence for non-steroid hormone transcription factors for the activation and up-regulation of L-plastin expression in prostate cancer].

OBJECTIVE: To search and identify the non-steroid receptor binding cis-acting elements in the L-plastin promoter in prostate cancer, and the correlative regulation pathway and transcription factors. METHODS: On the basis of construction of the L-plastin promoter luciferase vectors which were removed the steroid hormone receptor AR and ER binding elements, the promoter on the vector was nest-deleted by Exonuclease III and the relative luciferase plasmids were constructed. Transfected these twelve plasmids into prostate cancer cell line LNCaP under dihydrotestosterone-stimulated situation or not and test the intensity of luciferase, then we got the regulation message of every 200 bp part of the promoter in prostate cancer. After the analysis of relative programme, we got the possible regu- lation pathway of non-steroid hormone transcription factors. After removing the possible transcription factors binding site sequence by site-specific mutagenesis, the changes luciferase of activities proved our reasoning. RESULTS: We succeed in segmental deletion of the L-plastin promoter, and constructing the relative plasmids containing part L-plastin promoter on luciferase vector pGL3-basic. After testing the luciferase activities of constructed plasmids, we found the sequence from 206 to 1 of L-plastin promoter had significant luciferase activity. The software TRANSFECT showed that there were binding elements for transcription factors AP-4 at seq-198 to 192 and SP-1 at seq-54 to 41 on the short part promoter (206 to 1). The recombinant plasmids deleted the AP-4 and SP-1 binding elements had lower luciferase activity than the wild-type. CONCLUSION: There are some other non-steroid hormone pathway to regulate the expression of L-plastin except the steroid hormone pathway in prostate cancer. The main binding sites of the non-steroid hormone regulator lies in the sequence from 206 to 1. Transcription factors AP4 and SP-1 may up-regulated the expression of L-plastin by binding these sites.

Animals↗

[Spectral signal recognition based on wavelet packet analysis].

Automated recognition of spectral signal is an importance part of spectral analysis, which jnvolves comparing spectral signals under the same conditions and obtaining relation of chemistry constitution according to similarity by pattern recognition method and computer technique, and is useful to determine the nature of spectral signal and validate spectral signal. This paper takes spectral signal according with Lambert-beer' law as object, introduces basic theory and method of recognition of spectral signal in brief, normalizes spectral signal to decrease difficulty, introduces wavelet analysis theory, puts forward the method of wavelet packet analysis to gain features of spectral signal, obtains formula to compute feature vector and error vector by statistics method, then makes use of binary tree' mode distinguishing step by step for quick recognition, and finally gives an example to explain.

Algorithms↗

[Needle laparoscopic varicocelectomy].

OBJECTIVE: We evaluated our method and effects of needle laparoscopic varicocelectomy for the treatment of varicocele. METHODS: 72 patients (105 lateral) diagnosed varicocele were performed laparoscopic varicocelectomy under epidural combined intravenous anesthesia from Feb, 2003 to Apr, 2005. Two 2 mm incisions and one 5 mm incision were made on the midline of lower abdomen, by which two 2 mm trocars and one 5 mm trocar were introduced. Vessel-sealing device (Ligasure) was used to seal the internal spermatic veins. RESULTS: All operations were completed successfully. Mean operation time was 15 minutes unilateral and 21 minutes bilateral. The patients were hospitalized for 3 to 5 days after procedure. Follow-up was scheduled for 6 to 12 months and there was no recurrence. CONCLUSION: Needle laparoscopic varicocelectomy gives favorable effect with minimal invasion, rapid recovery, which is the best choice for the treatment of varicocele.

Adolescent↗

Restitution properties during ventricular fibrillation in the in situ swine heart.

BACKGROUND: Although restitution has been hypothesized to determine action potential duration (APD) during ventricular fibrillation (VF), cardiac memory may also be important. METHODS AND RESULTS: Transmembrane recordings were made with a floating microelectrode from the anterior right ventricular wall in 6 pigs during up to 60 seconds of VF. The recordings were divided into 5-second intervals, and APD60 and the diastolic interval (DI) were calculated for each activation cycle throughout each interval. Stepwise linear regression was used to determine how well each APD60 [APD60(n)] was predicted by the 4 previous DIs (n-1, n-2, n-3, n-4) and the 3 previous APD60s (n-1, n-2, n-3). A mean+/-SD of 3+/-1.5 of the variables entered the regression equation. DI(n-1) (70% of intervals) and APD60(n-1) (71% of intervals) appeared most frequently in the regression equations and were the first or second variables entered during the stepwise regression in 87% and 76% of the intervals in which they were present, respectively. The coefficients of DI(n-1) and APD60(n-1) were positive 89% and 98% of the time, respectively. R2 of the regression for all entered variables during all intervals was 0.39+/-0.05. CONCLUSIONS: The high incidence and positive coefficient of DI(n-1) indicate that restitution is important in determining APD during VF, whereas the similarly high incidence and positive coefficient of APD(n-1) indicate that cardiac memory is equally important. The finding that the regression equation accounts for only 39% of the variability of APD indicates that factors other than restitution and memory are also important in determining APD during VF.

Action Potentials↗