Empirical analysis on lumbar disc herniation treated with "sinew-bone three needling technique" of Chinese medicine / 中国针灸

The paper presents professor WU Han-qing's experience in treatment of lumbar disc herniation (LDH) with "sinew-bone three needling technique" of Chinese medicine. Based on the theory of meridian sinew, the points are located by "three-pass method" in terms of the distribution of meridian sinew and syndrome/pattern differentiation. The cord-like muscles and adhesion are relieved by relaxing technique to work directly on the affected sites and alleviate the local compression to the nerve root. The needle technique is operated flexibly according to the affected regions involved, due to which, the needling sensation is increased while the safety ensured. As a result, the meridian qi is enhanced, the mind and qi circulation is regulated; and the clinical effect is improved.

Experience in treatment of cervical spondylotic radiculopathy by relaxing the sinew of hand three / 中国针灸

The clinical experience of professor

Nine-hole Peg Test and Ten-meter Walk Test for Evaluating Functional Loss in Chinese Charcot-Marie-Tooth Disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The 9-hole peg test (9-HPT) and 10-meter walk test (10-MWT) are commonly used to test finger motor function and walking ability. The aim of this present study was to investigate the efficacy of these tests for evaluating functional loss in Chinese Charcot-Marie-Tooth (CMT) disease.</p><p><b>METHODS</b>Thirty-four Chinese CMT patients (CMT group) from August 2015 to December 2016 were evaluated with 9-HPT, 10-MWT, CMT disease examination score, overall neuropathy limitation scale (ONLS), functional disability score, and Berg Balance Scale (BBS). Thirty-five age- and gender-matched healthy controls (control group) were also included in the study. Student's nonpaired or paired t-test were performed to compare data between two independent or related groups, respectively. The Pearson test was used to examine the correlations between recorded parameters.</p><p><b>RESULTS</b>The mean 9-HPT completion time in the dominant hand of CMT patients was significantly slower than that in the healthy controls (29.60 ± 11.89 s vs. 19.58 ± 3.45 s; t = -4.728, P < 0.001). Women with CMT completed the 9-HPT significantly faster than men with CMT (dominant hand: 24.74 ± 7.93 s vs. 33.01 ± 13.14 s, t = 2.097, P = 0.044). The gait speed of the average self-selected velocity and the average fast-velocity assessed using 10-MWT for CMT patients were significantly slower than those in the control group (1.03 ± 0.18 m/s vs. 1.44 ± 0.17 m/s, t = 9.333, P < 0.001; 1.31 ± 0.30 m/s vs. 1.91 ± 0.25 m/s, t = 8.853, P < 0.001, respectively). There was no difference in gait speed between men and women. Both 9-HPT and 10-MWT were significantly correlated with the ONLS, functional disability score, and BBS (P < 0.05 for all).</p><p><b>CONCLUSION</b>The 9-HPT and 10-MWT might be useful for functional assessment in Chinese patients with CMT.</p>

Differences of the regulation on the expression of mucin 1 induced by two single-strand RNA viruses, respiratory syncytial virus and influenza virus / 病毒学报

To investigate whether influenza virus (IFZ) could up-regulate the expression of mucin 1 (MUC1) which exists in epithelial cells of upper respiratory track to restrict the inflammation, as respiratory syncytial virus (RSV) does. Quantitative RT-PCR and Western Blot were performed to detect the expression level of MUC1 induced by two single-strand RNA viruses in A549 cell lines. HEp-2 and MDCK cells were used respectively to culture RSV and IFZ. At 24h post A549 cells infection with the same titer of RSV or IFZ, the total RNA was harvest, qRT-PCR was then performed to observe the expression level of MUC1 mRNA. Meanwhile, at 24 h and 48 h post A549 cells infection with the same titer of RSV or IFZ, the total protein and supernatant were collected respectively after cell lysis, Western Blot was then used to detect the expression level of MUC1. Results showed that RSV could up-regulate the expression of MUC1 in airway epithelial cells with a significant dose-effect correlation, whereas IFZ could not. This study firstly investigated the differences of the regulation on the expression of MUC1 induced by two single-strand RNA viruses, and demonstrated initially that the mechanism of IFZ self-limiting differed from RSV, which attributed to up-regulation of the expression level of MUC1.

A new method for isolation and culture of tracheal epithelial cell of mice / 生理学报

Since most of the respiratory epithelial cell lines are descended from neoplastic tissues or have been fused with neoplastic cells when being selected to a cell line, their biological behaviors are far different from normal respiratory epithelial cells. Aiming at better reflecting the biological properties of epithelial cells under respiratory pathological conditions, our study probed into a new isolation technique and culture method of mice respiratory epithelial cell. Pronase was applied for cell isolation from mouse trachea, and a modified medium with collagen-coated plate for primary culture. The ciliary movement can be observed under microscope, which demonstrates that the original biological functions of the tracheal epithelial cell have been reserved with our method. The research presents the efficacy, convenience and reliability of the separation technique and culture method established by this study, and laying preferable condition for cell models of respiratory diseases. Meanwhile, this method may be used for other animal models.

Construction of different mutants of HA-tagged human RAGE gene and their eukaryotic expression / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct eukaryotic expression vectors for HA-tagged receptor for advanced glycation end products (RAGE) mutants.</p><p><b>METHODS</b>Site-directed mutagenesis was applied to wild-type RAGE gene cloned in the pcDNA3 vector with HA tag to obtain the mutants pcDNA3-HA-RAGE(S391A), pcDNA3-HA-RAGE(S399A), pcDNA3-HA-RAGE(S400A), and pcDNA3-HA-RAGE(T401A). After identification by sequencing, the mutants were transfected into HEK293 cells, and the expression of these mutants were detected by Western blotting using anti-HA antibody.</p><p><b>RESULTS</b>The HA-tagged RAGE mutants constructed were verified successfully by sequencing, and highly expressed in HEK293 cells.</p><p><b>CONCLUSION</b>The success in constructing HA-tagged RAGE mutants, which are highly expressed in eukaryotic cells, may facilitate the functional study of RAGE in cell signal transduction.</p>

BRCA1 regulates progesterone receptors A and B protein expressions in breast cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the regulatory role of BRCA1 in the expression of progesterone receptors A and B (PRA and PRB) in breast cancer cells.</p><p><b>METHODS</b>Breast cancer MCF-7 cells were transfected with pFlag-CMV2-BRCA1 wt plasmid containing a full-length BRCA1 cDNA or with BRCA1-specific siRNA via lipofectamine 2000 to induce overexpression or suppressed expression of BRCA1, respectively. Twenty-four hours after the transfection, the cells were incubated in fresh culture medium containing 100 nmol/L progesterone for 24 h. The total RNA extract or whole cell lysate was prepared for detecting BRCA1, PRA and PRB expressions using RT-PCR and Western blotting.</p><p><b>RESULTS</b>The protein expressions of PRA and PRB were significantly decreased whereas their mRNA expressions remained unchanged in MCF-7 cells overexpressing BRCA1. In MCF-7 cells with BRCA1 knock-down, in contrast, the PRA and PRB protein expressions were markedly increased.</p><p><b>CONCLUSION</b>In breast cancer cells, exogenous and endogenous BRCA1 can both down-regulate the expressions of PRA and PRB at the protein level.</p>

Phosphoprotein affinity profiling--a novel method for screening regulators of lipopolysaccharide signaling pathway / 南方医科大学学报

<p><b>OBJECTIVE</b>To To establish a method of phosphoprotein affinity profiling for identifying phosphoproteomic differences between Thp-1 cells with or without lipopolysaccharide (LPS) stimulation, aiming to screen potential regulators involved in LPS pathway.</p><p><b>METHODS</b>Thp-1 cells were stimulated with 100 ng/ml PMA for 48 h to induce differentiation into mature macrophages, which, after culture for another 48 h in the absence of PMA, were either treated with 100 ng/ml LPS for 30 min or left untreated. After desalting procedure with ultrafiltration, the phosphoproteins enriched by phosphoprotein metal affinity column (PMAC) of both groups were run on 2-D electrophoresis to find the spots with different phosphorylation status. Finally, some of these spots were identified by mass spectrometry (MS) and subsequent bioinformatic analysis.</p><p><b>RESULTS</b>Compared to untreated Thp-1 cells, LPS stimulated Thp-1 cells showed 29 spots with reproducible alterations on the 2-D map, including 8 representing up-regulated spots, 7 new spots, 10 down-regulated spots, and 4 absent spots. The newly emerged and absent protein spots were subjected to MS analysis, and 4 of them were identified to be involved in various cellular processes such as proteolysis, signal transduction and protein folding. Among these, phosphorylation of proteasome C2 subunit was dramatically up-regulated in LPS-stimulated cells, as was consistent with previous reports; the phosphorylation of Z-DNA-binding protein 1 has not been reported so far and needs further confirmation.</p><p><b>CONCLUSION</b>Phosphoprotein affinity profiling is an attractive method for screening novel regulators involved in LPS signaling pathways and can be widely used in systemic study of signal transduction.</p>

Construction and expression of the fusion vector for HA-tagged human RAGE gene / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a mammalian expression vector for HA-tagged receptor of advanced glycation end products (RAGE).</p><p><b>METHODS</b>Human RAGE cDNA codon region was amplified by PCR from human cDNA library and cloned into the pcDNA3 vector following routine procedures. After identification by enzyme digestion, PCR and sequencing, the correct recombinant vector RAGE/pcDNA3 was inserted with HA sequence behind the signal peptide sequence of RAGE. After identification by sequencing, HA-RAGE/pcDNA3 was transfected into HEK293 cells, and its expression was detected by Western blotting.</p><p><b>RESULTS</b>Identification by enzyme digestion, PCR and sequencing, confirmed the validity of the recombinant vector RAGE/pcDNA3, and HA-RAGE/pcDNA3 was highly expressed in HEK 293 cells.</p><p><b>CONCLUSION</b>HA-tagged RAGE is successfully constructed and expressed in mammalian cells, which may facilitate functional study of RAGE in cell signal transduction.</p>

Effect of p53 gene knockout on cell migration / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of p53 gene in serum-induced cell migration.</p><p><b>METHODS</b>The effects of p53 knockout on serum-induced formation of lamellipodia and cell migration were observed using Transwell cell migration system.</p><p><b>RESULTS</b>p53(+/+) cells developed lamellipodia upon serum stimulation and showed enhanced activity of cell migration, but these effects were not observed in p53 knockout cells after serum stimulation.</p><p><b>CONCLUSION</b>p53 plays a role in serum-induced cell migration.</p>