Professor YANG Ji-guo's experience in treatment of functional gastrointestinal diseases with acupuncture combined with umbilicus moxibustion / 中国针灸

Professor YANG Ji-guo's clinical experience in treatment of functional gastrointestinal diseases was summerized. Professor YANG Ji-guo believes that this disease is caused by the deficiency of six fu organs. Dysfunction of six fu organs in descending transportation is the basic pathogenesis. The principle of acupoint selection includes benefiting gastrointestinal functions, unblocking and purging six fu, soothing liver qi and calming down the mind. In treatment, acupuncture is combined with umbilicus moxibustion. In acupuncture, the deqi promoting technique by rotating and trembling needle is adopted. Focusing on the deficiency of six fu organs, umbilicus moxibustion is adopted to benefit the spleen and stomach and harmonize the functions of six fu organs for both biao (symptoms) and ben (root cause).

Analysis of genetic characteristics in two Chinese children of type Ⅱ Waardenburg syndrome / 中华耳鼻咽喉头颈外科杂志

Objective: To screen and analyze the mutations of MITF gene in two children of type Ⅱ Waardenburg syndrome (WS2) from different families in Yunnan,China,and to explore the possible molecular pathogenesis. Methods: With informed consent, medical history collection, physical examinations, audiological evaluation, and high resolution computer tomography (HRCT) scan of temporal bone were performed on the two WS2 probands and their family members. Genomic DNA was extracted from peripheral blood of all individuals. The coding regions including all exons, part of introns and promoters of MITF, PAX3, SOX10, SNAI2, END3, ENDRB, and KITLG genes were sequenced by high-throughput sequencing. According to the results of high-throughput sequencing, pathogenic mutations detected in the probands and their parents were verified by Sanger sequencing. Results: The proband 1 carried c.641_643delGAA mutation in the 7th exon of MITF gene, which was a frame-shift mutation resulting in an amino acid change of p.214delR. It was a de novo mutation as the parents of proband 1 showed no variation on this site. The proband 2 carried heterozygous loss of the large fragment ranging from exon 1 to exon 9 of MITF gene, which defected the function of MITF protein. Conclusion: Genetic examinations provide important evidence for diagnosis of Waardenburg syndrome. Heterozygous mutation c.641_643delGAA and heterozygous loss of the large fragment ranging from exon 1 to exon 9 of MITF gene might be the molecular pathogenesis of the two WS2 probands in this study.

Clinical Efficacy of Modified Zhibo Dihuangwan in Treatment of Purpura Nephritis with Deficiency of Liver and Kidney Yin in Children / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of modified Zhibo Dihuangwan on henoch-schonlein purpura nephritis with deficiency of liver and kidney yin in children (HSPN) and its effect on immune inflammatory response and hypercoagulable state. Method:Totally 120 patients were randomly divided into observation group (60 cases) and control group (60 cases) by random number table. Patients in two group was orally given prednisolone acetate tablets, 1.5-2 mg·kg-1·d-1, 2 times. Four weeks later, the drug was taken orally every other day, and the dosage decreased gradually after 4 weeks. Besides, patients in control group was intravenously dripped with cyclophosphamide, 8-12 mg·kg-1·d-1, for 2 days, and stopped for 2 weeks before another treatment course. The treatment lasted for 6 months. In the control group,Dabuyin Wan was taken orally,3 g/time,3 times/d.Patients in observation group was also added with modified Zhibo Dihuang Wan, 1 doe/day. The treatment lasted for 6 months. Urine routine was tested once a month, and disappearance time and rate of hematuria and albuminuria were recorded. The 24 h urine protein quantification, levels of microalbuminuria (mAlb) and urinary β2-microglobulin (β2-MG) were assessed before and after treatment. Furthermore, deficiency of liver and kidney Yin was scored, and levels of T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+), fibrinogen (FIB), D-dimer (D-D), fibrin degradation products (FDP), interleukin-2 (IL-2), interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-10 (IL-10) were detected. Result:The clinical efficacy in observation group was superior to that in control group (Z=2.078，P<0.05). Disappearance times of hematuria and albuminuria of children in observation group were shorter than those in control group (P<0.01). The disappearance rate of proteinuria in observation group was 90.48%（38/42）, which was higher than 69.77%（30/43） in control group (χ2=5.694，P<0.05). The 24 h urinary protein quantity, mAlb and levels of β2-MG, FIB, D-D and FDP in observation group were lower than those in control group (P<0.01). The levels of CD3+, CD4+, IL-2 and IFN-γ and the ratio of CD4+/CD8+ in observation group were higher than those in control group (P<0.05), while the CD8+, IL-4 and IL-10 were lower than those in control group (P<0.05). The efficacy in observation group was better than that in control group (Z=2.106，P<0.05). Conclusion:In addition to conventional western medicine therapy, modified Zhibo Dihuang Wan have an effect on HSPN with deficiency of liver and kidney Yin in children by promoting the disappearance of albuminuria and hematuria, shortening the course of disease, improving T lymphocyte subpopulation, reducing inflammatory reaction and correcting hypercoagulable state of blood, with better clinical efficacy and syndrome effect of traditional Chinese medicine.

Exploration of the Suitable Culture Conditions for Extracellular Microvesicles Derived from Human Mesenchymal Stem Cells / 中国实验血液学杂志

OBJECTIVE@#To explore the appropriate procedures for preparing extracellular microvesicles (MV) derived from human mesenchymal stem cells (MSC).@*METHODS@#Human MSCs from umbilical cords were cultured in a serum-free medium and maintained in a basal medium for 72 hours after the cell confluence reached to 80%. The supernatants of cultured cells were collected and MVs were enriched. MVs were identified by flow cytometry and electron microscopy. The total protein amount in MVs was used as a parameter for the content of MVs. The supernatants were adjusted to different pH values, and the output of MVs was detected. The supernatants were also collected for enriching the MV and detecting the protein content of MV after the cells were maintained in the basic medium for different time.@*RESULTS@#Flow cytometric analysis showed that the MVs expressed CD9, CD63 and CD81, morphologically presented round under an electron microscope and the diameter of MV was around 100 nm. After enrichment of MV, the protein content of MVs in the supernatants was 416.8±128.1, 255.4±77.9 and 142.8±46.4 μg per 10 MSC，respectively at pH of supernatant 3, 7 and 9 (P＜0.05). The protein content of the supernatants per 10 MSC was 173.6±44.5, 262.4±49.6 and 364.2±37.8 μg respectively after starvation culture for 48, 72 and 96 hrs (P＜0.05).@*CONCLUSION@#MVs can be readily collected after MSCs were starved for 96 hours, and the pH of the supernatants is adjusted at 3.0.

The Clinical Value of PELP1 for Breast Cancer: A Comparison with Multiple Cancers and Analysis in Breast Cancer Subtypes / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Proline, glutamic acid, and leucine-rich protein 1 (PELP1), a novel nuclear receptor (NR) co-regulator, is highly expressed in breast cancer. We investigated its expression in breast cancer subtypes, in comparison with other breast markers as well as cancers from different sites. Its prognostic relevance with different subtypes and other NR expression was also examined in breast cancers. MATERIALS AND METHODS: Immunohistochemical analysis was performed on totally 1,944 cancers from six different organs. RESULTS: PELP1 expression rate was the highest in breast cancers (70.5%) among different cancers. Compared to GATA3, mammaglobin and gross cystic disease fluid protein 15, PELP1 was less sensitive than GATA3 for luminal cancers, but was the most sensitive for non-luminal cancers. PELP1 has low expression rate (<20%) in colorectal cancers, gastric cancers and renal cell carcinomas, but higher in lung cancers (49.1%) and ovarian cancers (42.3%). In breast cancer, PELP1 expression was an independent adverse prognostic factor for non-luminal cancers (disease-free survival [DFS]: hazard ratio [HR], 1.403; p=0.012 and breast cancer specific survival [BCSS]: HR, 1.443; p=0.015). Interestingly, its expression affected the prognostication of androgen receptor (AR). AR(pos)PELP1(lo) luminal cancer showed the best DFS (log-rank=8.563, p=0.036) while AR(neg)PELP1(hi) non-luminal cancers showed the worst DFS (log-rank=9.536, p=0.023). CONCLUSION: PELP1 is a sensitive marker for breast cancer, particularly non-luminal cases. However, its considerable expression in lung and ovarian cancers may limit its utility in differential diagnosis in some scenarios. PELP1 expression was associated with poor outcome in non-luminal cancers and modified the prognostic effects of AR, suggesting the potential significance of NR co-regulator in prognostication.

Whether Fire-needle Therapy Benefits Plaque Psoriasis: A Multicenter, Randomized, and Controlled Trial / 中国结合医学杂志

OBJECTIVE@#To observe the clinical effectiveness and safety of fire-needle therapy, an external approach of Chinese medicine in treating plaque psoriasis.@*METHODS@#This study was a two-parallel-arm randomized controlled trial. A total of 151 participants with plaque psoriasis were randomly assigned to the fire-needle therapy group (treatment group, 76 cases) or the control group (75 cases) at a 1:1 allocation ratio using SAS software. All participants received Oral Huoxue Jiedu Decoction (, HXJDD) and applied externally vaseline cream twice a day. Participants in the treatment group received fire-needle therapy once weekly for 4 weeks plus HXJDD and vaseline cream applied the same as the control group. The primary outcome measure was Psoriasis Area and Severity Index (PASI) score, and the secondary outcomes were Dermatology Life Quality Index (DLQL), and Hamilton Anxiety Rating Scale (HAMA), as well as Chinese medicine (CM) syndrome score and photos of target lesions. The indices were evaluated before and after treatment.@*RESULTS@#Sixty-eight patients in each group completed the study. The treatment group has not yet achieved significant improvement in PASI score (P>0.05) compared to the control group. However, significant differences were found between the two groups in relieving CM syndrome (P<0.05) and improving quality of life (P<0.05).@*CONCLUSION@#Fire-needle appears to be safe and may have benefit for psoriasis, the short-term treatment and small sample size limit the conclusions of this study. Further rigorous randomized controlled trials with longer treatment are recommended.

Therapy approaches in treatment of recurrent ischemic angina with internal mammary artery bridge after coronary artery bypass grafting / 西安交通大学学报(医学版)

Objective To investigate the clinical therapeutic methods and their curative effects in recurrent ischemic angina for internal mammary artery resterosis after coronary artery bypass grafting (CABG).Methods We enrolled the patients who had recurrence of ischemic angina for restenosis of internal mammary artery graft after CABG as research subjects in the Affiliated Hospital of Yan'an University from January 2014 to January 2016.The 42 patients were divided into three groups according to the different treatment approaches for recurrence of ischemic angina:Group A (n=22)who received internal mammary artery interventional therapy;Group B (n=12)who received coronary artery bypass grafting treatment;and Group C (n=8)who received left subclavian artery proximal stent treatment.Then we compared the clinical therapeutic effects in the three groups.Results The success rate in Group C was 100%,which was the highest in the three groups,and the post-operative restenosis rate was 0.The hospitalization time was significantly shorter in Group A than in Group B (P＜0.05).However,the two groups did not significantly differ in mortality,success rate or restenosis rate (P＞0.05).Conclusion We should select the appropriate treatment according to the patient's specific situation for recurrent ischemic angina. Endovascular treatment has evident therapeutic effects,rapid postoperative recovery,and lower treatment risk, making it the preferred treatment when possible.

Chinese herbal medicine Euphorbia esula extract induces apoptosis and inhibits the proliferation, migration and invasion of multidrug resistant gastric carcinoma cells / 生物医学工程学杂志

This paper aims to study the effects of traditional Chinese medicine on multidrug resistant human gastric cancer cells in the cell proliferation, migration, invasion and apoptosis, and to study the apoptosis-inducing pathway. Different dilutions of extract were used to process human multidrug resistant gastric cancer SGC7901/ADR cells. Cell proliferation inhibition phenomenon was determined by MTT experiment. Nuclear morphological changes of apoptotic cells and apoptotic indexes were observed and determined by Hochest33528 staining followed with fluorescence microscope observing. Flow cytometry was used to detect cell apoptosis rate. Cell migration and invasion ability were observed and determined by Transwell method. Spectrophotometry was used to detect caspase-3 and caspase-9 enzyme activity. Western blotting was used to detect subcellular distribution of cytochrome c. The results showed that extract had obvious inhibition effect on proliferation of gastric cancer multidrug resistant SGC7901/ADR cells, which was time- and concentration-dependent. After processing multidrug resistant gastric cancer SGC7901/ADR cells with extract, the apoptotic index and apoptosis rate were significantly increased than those in the control group, which showed a time- and dose-dependent mode; but if a caspase inhibitor was added, apoptosis index was not obviously increased. Transwell method showed that migration and invasion ability of the extract-processed SGC7901/ADR cells dropped significantly. Spectrophotometry showed that in extract-processed SGC7901/ADR cells, caspase-3 and caspase-9 expression were increased, which had significant differences with the control group. Western blotting test showed that the distribution of cytochrome c decreased in mitochondria, while increased in the cytoplasm (i.e., cytochrome c escaped from mitochondria to the cytoplasm). In conclusion, extract could inhibit the proliferation, migration and invasion, and induce apoptosis in human gastric cancer multidrug resistant SGC7901/ADR cells; and cytochrome c, caspase-9 and caspase-3 might be involved in cell apoptosis induced by extract, suggesting endogenous or mitochondrial apoptotic pathway.

Progesterone Promotes Human Bone Marrow Mesenchymal Stem Cells to Synthesize Fibronectin via ERK Pathway / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate whether the progesterone can promote fibronection (FN) synthesis by human bone marrow mesenchymal stem cells (MSCs) and to explore the potential underlying mechanism.</p><p><b>METHODS</b>The human bone marrow MSCs were cultured in a serum-free medium with progesterone for 72 hours, the MTT test was performed to observe the proliferation status and adhension ability of the treated cells. Western blot was used to detect the content of FN in MSDs with GAPDH as the internal reference, the phosphorylation of ERK1/2, as well as the FN content in MSC treated by PD98059, a specific inhibitor of ERK1/2.</p><p><b>RESULTS</b>The progesterone at a range of certain doses not effect on the proliferation of human bone marrow MSCs. Progesterone (25 µg/L) treatment enhanced the FN expression and adherent ability of marrow MSCs. Progesterone could induce prompt phosphorylation of ERK 1/2 and its promoting effects on FN synthesis was reversed by PD98059.</p><p><b>CONCLUSION</b>The progesterone can promote FN synthesis by human bone marrow MSCs via ERK 1/2 pathway, and it might be used to culture MSCs in serum-free medium.</p>

Slowly-adhered primary bone marrow mesenchymal stem cells may exhibit more potent immuno-regulatory activity / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to investigate the relationship between the adherent ability of freshly isolated MSCs with their inhibitory effect on lymphocyte activation.</p><p><b>METHODS</b>Human bone marrow mononucleated cells were maintained in culture for 48 hours, the attached and the non-attached cells were then cultured separately and the adherent cells were collected and passaged. Cellular surface markers were analyzed with flow cytometry. Alkaline phosphatase activity and the intracellular lipid droplets were measured by histological staining, the in vitro osteogenesis and adipogenesis were identified. One-way mixed lymphocyte reaction was used to evaluate the suppressive activity of the adherent cells on lymphocyte proliferation, the prostaglandin E2 level in supernatant of cultured cells was detected by ELISA.</p><p><b>RESULTS</b>Some cells attached to the plastic after the bone marrow mononucleated cells were allowed to adhere for 48 hours. The slowly-attached cells were fibroblast-like in morphology, homogenously positive for CD44 and CD73 and negative for CD31 and CD45. They could be coaxed into osteoblasts and adipoblasts under the standard inductive conditions. These cells were able to inhibit lymphocyte proliferation in mixed lymphocyte reaction and their effect was more potent than those from the adherent cells appeared within 48 hours. The concentration of prostaglandin E-2 in the supernatants of the slowly-adhered cells was significantly higher than that in the MSCs cultured with the traditional method (90.8 ± 10.37 ng/ml vs 70.2 ± 8.98 ng/ml) (P < 0.01).</p><p><b>CONCLUSIONS</b>The MSCs exist in the marrow mononucleated cells after adherent culture for 48 hours, and the MSCs may exhibit more potently inhibitory activity on lymphocyte activation.</p>

Research progress on epithelial-mesenchymal transition in cancer recurrence and metastasis / 浙江大学学报·医学版

Epithelial-mesenchymal transition (EMT) is a process in which epithelial cells lose their morphology and function and gradually transformed into mesenchymal-like cells. It is considered that EMT is the main cause for tumor recurrence and metastasis. Many factors are involved in the regulation of EMT, such as E-cadherin, transforming growth factor-β, Wnt signaling pathway, microRNA and EMT-related transcription factors. This article reviews the research progress on EMT and the involved mechanisms, and thus to provide a new perspective on cancer therapy in the future.

Putrescine Promotes Human Marrow Mesenchymal Stem Cells to Differentiate along Osteogenic Pathway / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of putrescine on the growth and differentiation of human bone marrow mesenchymal stem cells (MSC) to develop a new inductive medium mixture for their osteogenic differentiation.</p><p><b>METHODS</b>Human bone marrow MSC were collected from three healthy donors and were used to observe the growth-promoting activity of putrescine with MTT test. Experiments were divided into 3 groups: (1) putrescine group, (2) positive control group (presence of dexamethasone, ascorbate, and glycerol phosphate) and negative group (d-alpha with 5% FCS). The cellular expression level of Runx-2 was detected by PCR assay after the culture was maintained for 1 week. After 2 weeks, the intracellular activity of alkaline phosphatase was revealed by histochemistry staining, the phosphatase activity, and the protein concentration in the cell lysates were also detected. Furthermore, MSC were cultured in the presence of putrescine for 2 weeks and Oil-red O staining was performed to reveal the differentiated adipocytes; the cells induced by the standard agent cocktail were used as the positive control.</p><p><b>RESULTS</b>Putrescine promoted the proliferation of human marrow MSC in a dose-dependent manner. MSC exposed to putrescine at a concentration of 100 µmol/L for 1 week expressed greatly higher level of Runx-2, compared with the negative control. Alkaline phosphatase activity was evidently observed after MSC were maintained in the presence of putrescine for 2 weeks. The phosphatase activity contrasted to the protein content in putrescine-treated MSC was significantly higher than that of the control cells (0.87±0.012 vs 0.52±0.010) (P<0.01), and also greatly higher than that of the positive control (0.83±0.029) (P=0.02). Oil red O staining showed that MSC treated by putrescine did not differentiate into adipoblasts.</p><p><b>CONCLUSION</b>Putrescine can promote the proliferation and osteogenic differentiation of MSC, suggesting the potential application of putrescine as a novel inductive agent for in vitro osteogenesis of MSC.</p>

Exploration of conditions for releasing microvesicle from human bone marrow mesenchymal stem cells / 中国实验血液学杂志

The release of microvesicles(MV) is one of the critical mechanisms underlying the angiogenesis-promoting activity of mesenchymal stem cells(MSC). This study was aimed to explore the appropriate condition under which MSC releases MV. Bone marrow samples from 5 healthy adults were collected, and MSC were isolated, culture-expanded and identified. MSC at passage 5 were suspended in medium without or medium with 10% fetal(FCS) calf serum and seeded into culture dishes. The culture was separately maintained in hypoxia (1% oxygen) or normoxia (around 20% oxygen), and 20 dishes of cells (2×10(6)/dish) were used for each group. The supernatants were collected for MV harvesting. The cell number was counted with trypan blue exclusion test and the protein contents in the MV were determined. MV were identified by observation under an electron microscope. The surface markers on MV were analyzed by flow cytometry. MTT test was performed to observe the pro-proliferative activity of MV that were added into the culture of human umbilical cord vein endothelial cells at a concentration of 10 µg/ml. The results showed that the majority of MV released by MSC were with diameters of less than 100 nm, and MV took the featured membrane-like structure with a hypodense center. They expressed CD29, CD44, CD73 and CD105, while they were negative for CD31 and CD45. The increase multiples of the adherent trypan blue-resistant cells cultured in normoxia with serum, in normoxia without serum, in hypoxia with serum and hypoxia in the absence of serum were 4.05 ± 0.73, 1.77 ± 0.48, 5.80 ± 0.65 and 3.69 ± 0.85 respectively, and the estimated protein contents per 10(8) cells were 463.48 ± 138.74 µg, 1604.07 ± 445.28 µg, 2389.64 ± 476.75 µg and 3141.18 ± 353.01 µg. MTT test showed that MV collected from MSC in hypoxia seemed to promote the growth of endothelial cells more efficiently than those from cells in normoxia. It is concluded that hypoxia can enhance the release of microvesicles from MSC, and cultivation of MSC in hypoxia and medium without serum may provide an appropriate condition for MV harvesting.

Antioxidative effects of cysteinyl leukotriene receptor antagonists montelukast and HAMI 3379 on ischemic injury in rat cortical neurons in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the antioxidative effects of two cysteinyl leukotriene receptors antagonists (CysLT1R and CysLT2R) montelukast and HAMI 3379 on ischemic injury of rat cortical neurons in vitro.</p><p><b>METHODS</b>Cultured rat cortical neurons were pretreated with CysLT1R antagonist montelukast and CysLT2R antagonist HAMI 3379, and then exposed to oxygen-glucose deprivation/recovery (OGD/R）or H2O2. Reactive oxygen species (ROS) mitochondrial membrane potential (MMP) depolarization, neuronal viability and lactate dehydrogenase (LDH) release were determined. Meanwhile, RNA interference was used to inhibit the expression of CysLT1R and CysLT2R，and the effects were observed.</p><p><b>RESULTS</b>ROS production in neurons was significantly increased after 1 h OGD, which reached the peak at 30 min and lasted for 1.5 h after recovery. Montelukast and HAMI 3379 at 0.01-1μmol/L moderately decreased OGD/R-induced ROS production (P<0.05). Montelukast mildly attenuated OGD/R-induced MMP depolarization (P<0.05)，but HAMI 3379 had no effect. H2O2 reduced neuronal viability and increased LDH release, namely inducing neuronal injury. Montelukast and HAMI 3379 at 0.1-1μmol/L moderately attenuated H2O2-induced neuronal injury (P<0.05). However, both CysLT1R siRNA and CysLT2R shRNA did not significantly affect the responses mentioned above.</p><p><b>CONCLUSION</b>In ischemic neuronal injury, montelukast and HAMI 3379 exert a moderate antioxidative effect, and this effect may be receptor-independent.</p>

Effect of histone deacetylase inhibitor NL101 on rat neurons / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the protective effect of histone deacetylase inhibitor NL101 on L-homocysteine (HCA)-induced toxicity in rat neurons, and the toxic effect on normal rat neurons.</p><p><b>METHODS</b>In the presence of NL101 at various concentrations, HCA (5 mmol/L)-induced changes in cell density, necrosis, and viability were determined in the mixed cultures of rat cortical cells and the primary cultures of rat neurons. The direct effect of NL101 on primary neurons was also observed in the absence of HCA. Histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) was used as the control. After the treatments, cell viability, the density, and morphology of neurons and glial cells, and cell necrosis were determined.</p><p><b>RESULTS</b>In the mixed cultures of cortical cells, NL101 had no effect on HCA (5 mmol/L)-induced cell number reduction at 0.001-10μmol/L; however, it significantly attenuated necrosis at 1-10 μmol/L, and increased neuronal number at 1 μmol/L. NL101 had no effect on the mixed cortical cells in the absence of HCA. In the primary neurons, NL101 reduced neuronal viability and mildly increased necrosis at 1-10 μmol/L in the absence of HCA, while it significantly attenuated HCA-induced neuronal viability reduction at 0.01-10 μmol/L and reduced neuronal necrosis at 1-10 μmol/L. The effects of NL101 were apparently similar to those of SAHA.</p><p><b>CONCLUSION</b>NL101 has protective effect on HCA-induced neuronal injury but it is neurotoxic at high concentrations, which is similar to the typical histone deacetylase inhibitor SAHA.</p>

Effects of the water channel aquaporin 4 deficiency on bleomycin-induced lung fibrosis in mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the effect of water channel aquaporin 4 (AQP4) on bleomycin-induced lung fibrosis in mice.</p><p><b>METHODS</b>In wild type and AQP4 gene knockout (AQP4-/-) mice, lung fibrosis was induced by injection of bleomycin (3 mg/kg) into the trachea and saline injection was used as a control. At d3, 7, 14, 28 after bleomycin-treatment, mice were randomly sacrificed in batch and the lung coefficient was determined. Serum levels of TGF-β1 and TNF-α were measured by ELISA and hydroxyproline contents in lung tissue were determined by Alkaline hydrolysis method. H-E staining and Masson's staining were performed to examine the pathological changes of lung tissues after bleomycin-treatment.</p><p><b>RESULTS</b>On d14 after bleomycin-treatment, the lung coefficients in wild type mice and AQP4-/- mice were 1.9-fold (12.69 ± 6.05 vs 6.80 ± 0.82, q=4.204, P<0.05) and 2.3-fold (14.05 ± 5.82 vs 6.05± 0.58, q=5.172, P<0.01) of that in control, respectively, but no significant difference was found between wild type and AQP4-/- mice in the lung coefficient value (P>0.05). The hydroxyproline contents in the lung increased after bleomycin-treatment; on d28, the lung hydroxyproline contents in wild type and in AQP4-/- mice were 1.55-fold (0.85 ± 0.22 g/mg vs 0.55 ± 0.14 μg/mg, q=4.313, P<0.05) and 1.4-fold (0.84 ± 0.13 μg/mg vs 0.60 ± 0.14μg/mg, q=4.595，P<0.05) of that in control, respectively, but no significant difference was noticed between wild type and AQP4-/- mice in lung hydroxyproline contents. There was a tendency that serum TGF-β1 and TNF-α levels increased in bleomycin-treated mice, but no significant difference was found between wild type and AQP4-/- mice. AQP4-knockout showed no effects on pathological changes of lung tissues with H-E staining and Masson's staining in mice with bleomycin-induced lung fibrosis.</p><p><b>CONCLUSION</b>AQP4 might not be involved in bleomycin-induced lung fibrosis in mice.</p>

Effects of leukotriene D4 on proliferation and migration of lung epithelial A549 cells in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of cysteinyl leukotriene (CysLT) receptor agonist leukotriene D4 (LTD4) on proliferation and migration in lung epithelial A549 cells.</p><p><b>METHODS</b>The expression of CysLT1 receptor and CysLT2 receptor was determined by immunofluoresence staining in A549 cells. A549 cells were treated with LTD4 (0.01-100 nmol/L) for 24-72 h. Cell viability was detected by MTT reduction assay. Cell migration was determined by modified scratch and healing model.</p><p><b>RESULTS</b>In A549 cells, CysLT1 receptor and CysLT2 receptor were mainly expressed in the cytoplasm, membrane and few in the nuclei. The treatment of LTD4 (0.01-100 nmol/L) for 24-72 h caused no effect on cell viability (Ps>0.05); when A549 cells were treated with 100 nmol/L LTD4 for 24, 48 and 72 h the cell viability was (103.00±4.46)%，(107.00±9.45)% and (105.00±9.02)% of control, respectively (Ps>0.05). The migration rate of A549 cells after scratching during the first 24 h was markedly greater than that during the second and third 24 h in the same concentration groups; however, no significant difference in migration rate was noticed when the cells were treated with different concentrations of LTD4 (0.01-100 nmol/L)(Ps>0.05). The migration of A549 cells was 1.15-fold, 1.21-fold and 1.06-fold of that of control when the cells were treated with 100 nmol/L LTD4 for 24, 48 and 72 h, respectively (Ps>0.05).</p><p><b>CONCLUSION</b>The proliferation and migration of A549 cells are not changed when treated with 0.01-100 nmol LTD4 for up to 72h.</p>

Research advances on the roles of nicotinamide phosphoribosyltransferase in inflammation / 浙江大学学报·医学版

Nicotinamide phosphoribosyltransferase (Nampt) is also called visfatin or pre-B-cell colony-enhancing factor. The functions of Nampt have been reported as a cytokine, an adipokine and the rate-limiting enzyme in nicotinamide adenine dinucleotide biosynthesis. As a pleiotropic multifunctional protein, Nampt is involved in a variety of physiological and pathological conditions including innate immunity, metabolic disorders, and stress; and Nampt also participates in inflammatory disorders such as acute lung injury, atherosclerosis, myocardial infarct, obesity, type 2 diabetes, and rheumatoid arthritis. The studies indicate that Nampt might be a potential target for pharmacological intervention against inflammatory diseases. We review research advances on the roles of Nampt in inflammation.

Analysis on clinical features and treatment of herpes zoster patients hospitalized in real world / 中国中药杂志

From the hospital information system (HIS) of 20 national grade III-A general hospitals, 2 960 cases of herpes zoster as the research object, analyzes the relations between the general information, syndrome of traditional Chinese medicine (TCM), western medicine combined diseases, the relationship between the solar term and the incidence of herpes zoster, and the combined use of Chinese and western medicine. Among the patients with 46-65 year old has the highest percentage of diseased; admission to general outpatient clinic is the most; the most common medical payment is medicare; combined disease such as hypertension, diabetes and coronary heart disease is more common; early treatment effect of herpes zoster is better than the sequelae; summer and autumn solar term patients is hospitalized more, TCM syndrome is damp heat of liver fire; about drugs, western medicine is the most commonly used vitamin B1 and mecobalamin, traditional Chinese medicine is the most frequently used Danhong injection, combination therapy with promoting blood circulation drugs and neurotrophic drugs. Thus, herpes zoster, more common in elderly patients, with no obvious relationship between solar term, should be early diagnosis and early treatment, often with combination of Chinese traditional and western medicine treatment.

Mechanisms of cholic acid for reducing damage to human live cell lines L-O2 induced by amanita toxic peptides / 中华实用儿科临床杂志

Objective To explore the mechanisms of different cholic acid for reducing damage to human liver cells lines L-O2 induced by amanita toxic peptides (amataxins).Methods According to different concentrations of amataxins,the experiment was conducted with different dosages in 5 groups:0.00 g/L,0.26 g/L,0.40 g/L,1.40 g/L and 2.80 g/L.The human liver cells lines L-O2 in the exponential growth phase were cultured into 96-well plates,1 ×103 cells per well After 24 hours,the concentrations of amanita toxic peptides mentioned above were added.The minimum concentration of mushroom toxins keeping the liver cells alive was determined after 24,48 and 72 hours,respectively,and MTT method was used to test each group's liver cell activity.The experiment included 3 groups:the control group,the damage group,and the cholic acid group.Each group had 3 time points:24,48 and 72 hours after being attacked.Twenty four hours after attack,in cholic acid group,cholic acid drugs including taurocholic acid gca,goose deoxycholic acid,gansu ammonia goose deoxycholic acid and bovine goose deoxycholic acid were given,respectively,to protect the injured liver cells.Cells' morphology changes were observed under the inverted phase contrast microscope,living cells were counted by using MTT method,and aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in the culture supernatant were tested by the biochemical method.Results The minimum attack concentration of lamanita toxic peptides keeping liver cell survival in vitro was 1.40 g/L.Seventy-two hours after attack by amanita toxic peptides,the absorbance value was 0.812 ± 0.035,0.345 ± 0.021,0.363 ± 0.018,0.387 ± 0.027,0.431 ± 0.018,0.465 ± 0.015 and 0.452 ± 0.030,respectively in the control group,the damage group,the taurocholic acid group,the goose deoxycholic acid group,the glycocholic acid group,the glycochenodeoxycholic acid group and the sodium deoxycholic acid group.Compared with the damage group,absorbance value 72 hours after attack in each cholic acid group gradually increased,and compared with damage group,the differences were statistically significant among goose deoxycholic acid group,glycocholic acid group,glycochenodeoxycholic acid group and sodium deoxycholic acid group(P ＜ 0.05).AST and ALT activity in each cholic acid group declined,and that in glycochenodeoxycholic acid group was the lowest.Compared with the damage group,the difference was statistically significant (P ＜ 0.01).Conclusions Cholic acid can protect human liver cells from the damage induced by amanita toxic peptides.Such effect may be related to the fact that both amanita toxic peptides and cholic acid are the substrates of NTCP and OATP.