Effects of repair for defect after xanthelasma removal with a orbicularis oculi myocutaneous flap by converse shift grafting / 中华医学美学美容杂志

Objective:To explore the method and effect of blepharoplasty of upper eyelid combined with orbicularis oculi myocutaneous flap in repair of the canthus defect after excision of xanthelasma.Methods:According to the relaxation of the upper eyelid skin and the width of the xanthoma to be removed, the double eyelid line and the peeling line were designed individually, and the myocutaneous flap was removed to repair the inner canthus defect. After resection of the xanthelasma, the orbicularis oculi myocutaneous flap was transferred retrogradely to repair the defect, and then the upper eyelid plasty was performed.Results:During the follow-up of 3-12 months after operation, 13 cases (24 eyes) of the transferred flaps in the inner canthus survived well, the incision scars were hidden, and the double eyelids on both sides were naturally symmetrical.Conclusions:The method of upper eyelid plasty combined with the reverse transfer of orbicularis oculi myocutaneous flap provides more options for the treatment of defects after resection of xanthoma of the inner canthus. The flap survives well, improves the shape of the upper eyelid while completely removing the focus, and avoids complications such as postoperative deformation and obvious scars.

Pathogenesis of Gouty Arthritis and Intervention with Chinese Medicines： A Review / 中国实验方剂学杂志

Gouty arthritis （GA） is the metabolic rheumatism caused by purine metabolism disorder， which can be acute or chronic. The main manifestations of GA include recurrent redness， swelling， heat pain， and dysfunction of the affected joints. According to the theory of modern medicine， GA is closely associated with the increase in uric acid， the participation of inflammatory cytokines， the weakening of antioxidant response， apoptosis， and the imbalance of intestinal flora and bone metabolism， whereas the specific pathogenesis remains unclear. GA is characterized by easy diagnosis， difficult treatment， and high recurrence rate， which seriously affects the life quality of patients. Colchicine， corticosteroids， non-steroidal anti-inflammatory drugs， and selective cyclooxygenase-2 inhibitors are the commonly used western medicines for this disease， which demonstrate remarkable short-term therapeutic effect. However， long-term use of these medicines will bring serious adverse reactions. Chinese medicines， with high safety and causing few adverse reactions， have a variety of active components which can act on multiple pathways and targets to exert synergistic effects， thus attracting wide attention. This paper systematically reviews the literature reporting the Chinese medicines in improving antioxidant response， reducing chondrocyte apoptosis， and regulating intestinal flora and bone metabolism， aiming to further clarify the pathogenesis of GA and provide a scientific basis for the clinical application of Chinese medicines in the prevention and treatment of GA.

Clinical Research on Treatment of Benign Prostatic Hyperplasia with Traditional Chinese Medicine： A Review / 中国实验方剂学杂志

Benign prostatic hyperplasia （BPH）， a common disease in urology and andrology， is mainly manifested as enlarged prostate glands， bladder outlet obstruction， and lower urinary tract symptoms（LUTS）， which seriously affects the quality of life of middle-aged and elderly men. This disease falls into the categories of "retention of urine" and "prostatic hypertrophy" in traditional Chinese medicine （TCM）. In recent years，many doctors have put forward their understandings of BPH based on academic classics and their clinical experience. Clinical research on the treatment of BPH with TCM has become increasingly abundant. The basic pathogenesis of BPH lies in the disturbance of Qi transformation in the bladder and poor blood circulation due to kidney Qi deficiency in the aged. The disease is located in the kidney and the bladder and is related to the dysfunction of the lung and the kidney. It is basically characterized by deficiency in origin and excess in superficiality. A large number of clinical research reports have proved that TCM is efficient in alleviating the clinical symptoms of BPH patients， improving their quality of life， reducing the volume of the prostate， and decreasing postoperative complications. In addition， the external treatment methods of TCM， such as acupuncture therapy， moxibustion， hot water bathing， acupoint application， anal suppository， and enema therapy， are also widely used in clinical practice， demonstrating the diverse ways of TCM in treating BPH. TCM and western medicine complement each other's advantages in the treatment of BPH， thus enhancing the clinical efficacy and reducing the occurrence of long-term complications. This study reviewed the etiology， pathogenesis， and treatment progress of BPH with TCM in recent years， and summarized the current research status. From three aspects of producing high-quality clinical research， standardizing the clinical diagnosis and treatment of TCM， and combining cutting-edge research to explore the mechanism of TCM， it provided suggestions for clinical research on the treatment of BPH with TCM to promote the development and application of TCM in the treatment of this disease.

Research progress in mechanical barrier function of intestinal mucosa in nonalcoholic fatty liver disease / 国际生物医学工程杂志

Intestinal mucosal barrier is the first line of defense against exogenous substances. A complete mechanical barrier can separate the body from the contents of the intestines and prevent the translocation of bacteria and microorganisms in the intestines, playing an important role in maintaining the homeostasis of the human body. If the intestinal mechanical barrier function is damaged, it will cause the intestinal endotoxin and bacterial translocation, leading to systemic inflammatory response syndrome and multiple organ dysfunction diseases. The barrier function of intestinal mucosa is a key link in the pathogenesis of nonalcoholic fatty liver disease (NAFLD). Therefore, the maintenance of intestinal mucosal barrier function is of great value for the diagnosis and treatment of NAFLD. In this paper, the research progress in intestinal mechanical barrier function of NAFLD was reviewed, and the main factors maintaining intestinal permeability of NAFLD were introduced, such as apoptosis, autophagy, apical junctional complex, cytoskeleton, main signal pathway, to provide reference for the diagnosis and treatment of NAFLD.

Effect of Zelen' s Guige Lingxian Decoction Combined with Moxibustion at Neck Jiaji Acupoint in Treatment of Cervical Spondylotic radiculopathy of Wind-cold-dampness Arthralgia Syndrome / 中国实验方剂学杂志

Objective::To investigate the clinical efficacy and mechanism of Guige Lingxian decoction combined with moxibustion at neck Jiaji acupoint on cervical spondylotic radiculopathy (CSR) of wind-cold-dampness arthralgia syndrome. Method::Totally 120 cases of cervical spondylotic radiculopathy with wind-cold-dampness syndrome were randomly divided into two groups: traditional Chinese medicine(TCM) group and combination group, with 60 cases in each group. The TCM group was treated with Guige Lingxian Tang 150 mL/time, twice a day. The combination group was treated with moxibustion at neck Jiaji acupoint in addition to the therapy of the TCM group, 30 minutes/time, once a day. Both groups were treated for 8 weeks. CSR 20 subscale score, SF-36 scale score, typical symptoms and signs remission time were recorded. The total effective rate and the cure rate were recorded after treatment. Serum levels of leukotriene B4(LTB4), leukotriene C4(LTC4), leukotriene D4 (LTD4), chitinase protein 40 (YKL-40), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were measured by enzyme linked immunosorbent assay(ELISA) before and after treatment. Result::The total effective rate was 98.11%(52/53), and the cure rate was 43.39%(23/53) in the combination group, which were higher than 91.04%(61/67) and 20.89%(14/67) in the TCM group (P<0.05). Compared with the TCM group, the scores of CSR 20 subscale and SF-36 scale in combination group were increased (P<0.05), and the remission time of typical symptoms and signs was shortened. Serum levels of LTB4, LTC4, LTD4, YKL-40, IL-1β and TNF-α were decreased (P<0.05). Conclusion::Moxibustion at neck Jiaji acupoint combined with Guige Lingxian decoction is effective in treating CSR of wind-cold-dampness arthralgia syndrome, which is worthy of clinical promotion. Its mechanism may be related to the inhibition of the expression of LT, and the reduction of the content of IL-1β and TNF-α in serum, thus promoting the repair of cervical cartilage and increasing the pain threshold.

Effect of Bushen Kangshuai Tablets on Chronic Fatigue Syndrome with Syndrome of Spleen and Kidney Deficiency / 中国实验方剂学杂志

Objective: To observe the effect of Bushen Kangshuai tablets combined with abdominal massage on chronic fatigue syndrome (CFS) with syndrome of spleen and kidney deficiency, and investigate its antioxidant stress and immune regulation effects. Method: One hundred and sixty patients were randomly divided into control group and observation group by random number table, 80 cases in each group. Patients in control group got abdominal massage, 1 time/day, 5 times/week, for a total of 3 treatment courses, 15 times in one course. Based on the treatment in control group, patients in observation group also received Bushen Kangshuai tablets, 6 tablets/time, 3 times/days, for a total of 12 weeks in treatment. Before and after treatment, symptom scores of spleen and kidney deficiency, fatigue scale-14 (SF-14), self-rating anxiety scale (SAS), self-rating depression scale (SDS) and Pittsburgh sleep quality index (PSQI) were graded; and levels of serum immunoglobulin (IgG, IgA and IgM), natural killer cells (NK), T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+/CD8+), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected. Result: In the rank sum test, the clinical efficacy in observation group was better than that in control group (Z=1.982, PPPP+, CD4+, CD4+/CD8+ were also higher then those in control group (PP+ (PPConclusion: Bushen Kangshuai tablets combined with abdominal massage to CFS with syndrome of spleen and kidney deficiency can significantly relieve fatigue, improve spleen and kidney deficiency, and improve immune function and antioxidant stress, superior to abdominal massage therapy alone in the clinical Efficacy.

Effect of Modified Guipitang Combined with Xingnao Kaiqiao Acupuncture in Treatment of Heart and Spleen Insufficiency Syndrome of Non-dementia Vascular Cognitive Impairment / 中国实验方剂学杂志

Objective: To explore the curative effect and partial mechanism of modified Guipitang combined with Xingnao Kaiqiao acupuncture in the treatment of non-dementia vascular cognitive impairment (VCIND). Method: Totally 122 patients with VCIND admitted to the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine (TCM) from May 2017 to October 2018 were randomly divided into modified decoction group (39 cases), acupuncture group (42 cases) and combination group (41 cases). All of the three groups were orally given routine anticoagulants and lipid-lowering drugs. The decoction group was orally given modified Guipitang 150 mL/times, 2 times/day, in addition to the routine treatment, the acupuncture group was treated with Xingnao Kaiqiao acupuncture method in addition to the routine treatment, involving Shuigou, Neiguan (bilateral), Sanyinjiao (bilateral), Sishencong, Xuanzhong (bilateral) and Taixi (bilateral) acupoints, 2 times/day, for six days a week, the combined group was treated with Xingnao Kaiqiao acupuncture in addition to modified Guipitang. All of the three groups were treated for 8 weeks. The Montreal cognitive assessment scale (MoCA scale, Beijing version) and activity of daily life (ADL) scale, TCM symptoms and clinical efficacy were scored before treatment, 4 weeks after treatment and 8 weeks after treatment in three groups. Serum levels of calcitonin gene-related peptide (CGRP) and cone-like protein-1 (VILIP-1) were measured by enzyme-linked immunosorbent assay at different time points. Result: Compared with decoction group and acupuncture group, MoCA score, ADL score and TCM syndrome score of the combined group were decreased, the total effective rate was increased significantly after 4 and 8 weeks of treatment, the serum CGRP content was increased, and the VILIP-1 content was decreased. Conclusion: Modified Guipitang combined with Xingnao Kaiqiao acupuncture has a definite curative effect on VCIND with heart and spleen insufficiency syndrome. Its mechanism may be related to the expansion of blood vessels, the alleviation of blood supply of brain and the reduction of neuron injury.

Accordion technique in the treatment of tibial delayed union or nonunion / 中华骨科杂志

Objective To assess the clinical curative effect of accordion technique in the treatment of tibial delayed union or nonunion.Methods From February 2016 to December 2017,data of 11 patients with tibial delayed union (n=8) or non-union (n=3) who had been treated by accordion technique with an Ilizarov ring external fixator were retrospectively analyzed.10 males and 1 female were included in our study,with an average age of 41.9 years (range,21-63 years).There were 5 cases of docking site delayed union after Ilizarov transport for chronic tibial osteomyelitis.There were 3 cases of fracture site delayed union after external fixation for open tibia comminuted fracture.There were 3 cases of nonunion after tibia closed fracture,including 2 cases who had hybrid external fixation treatment,and 1 case who had conservative treatment.All the cases received accordion technique using Ilizarov ring external fixators.First,gradual compression at the fracture site was conducted until the bony contact was seen on a radiograph.After bony contact,compression was continued at a rate of 0.85 mm/d for a week,followed by distraction of 0.85 mm/d for 2-3 weeks.Afterward,a second compression was conducted using same rate and time with the distraction procedure.And there was a 7-day latent period between compression and distraction.One or two cycles of compression-distraction were needed before union was present radiographically.Results Bony union was obtained in all 11 patients after a mean time of 5.4 months (from 3 to 9 months).The mean follow-up for the 11 patients was 15.2 months (from 11 to 29 months).The mean duration of the accordion technique treatment was 50.2 d (range,35-67 d).The accordion technique was used 1 time for 8 patients,and 2 times for 3 patients.The mean duration of bone consolidation was 114.9 d (range,64-239 d).According to Paley evaluation criteria,osseous results were excellent in 8 cases,good in 3 cases,with a good to excellent rate of 100％(11/11);functional results were excellent in 7 cases,and good in 4 cases,with a good to excellent rate of 100％(11/11).Conclusion The accordion technique is a minimal invasive,safe and reliable treatment program for tibial delayed union or nonunion.

Effectiveness of remold eyelid structure method of blepharoplasty for thickening eyelid / 中华医学美学美容杂志

Objective To explore the method and efficacy of using "Y" flap formed to remold double eyelid structure of blepharoplasty for thickening eyelid.Methods A total of 97 cases of single eyelids and thickening eyelid were treated with surgery,to release the space before tarsus fascia and levator palpebrae aponeurosis to form a continuous interspace of the orbicularis oculi muscle.Then joint fascia was reconstructed to format "Y" flap,on which both sides of the levator aponeurosis and orbital septum fascia were tightened and the redundant fat and fascial organization removed.The lateral fat pad under the orbicular muscle were wedge-shaped sheared along the orbital margin after double eyelid was formed.Few patients presented acrimal glands prolapse,which suspended to return the lower margin of the orbital bone.Finally according to the upper eyelid anatomy suture each layer of incision was closed.Results The 97 cases underwent surgical treatment and the incisions were healed by first intention,in which 3 cases had slight skin ptosis at the lateral double crease due to the conservative tissue removal and 2 cases had the double eyelid width asymmetry due to the tissue was removed unevenly.They were adjusted satisfiedly 3 months later.All cases were followed up for more than 3 months and acquired satisfactory improvement and beautification.Incision was healed hiddenly.Conclusions This method can be used to tease and adjust the upper eyelid levels clearly and completely,which makes redundant tissues removed in place and double eyelid located accurately.By this method single eyelid combined thickening eyelid could be solved aestheticly at the same time and get a natural,popular and natural shape.

Effect of PIVKA-Ⅱ and AFP levels on the prognosis of patients with HBV infection-related hepatocellular carcinoma / 中国实用内科杂志

OBJECTIVE: To evluated the prognostic potential of preoperative PIVKA-Ⅱ and AFP to the patients of HBV infection-related hepatocellular carcinoma(HCC) after radical resection. METHODS: Chronic HBV infection-related HCC patients who undergone resection in the Affiliated Cancer Hospital of Zhengzhou University from 2009 to 2013 with competed data of clinical information,laboratory results and follow-up records were enrolled our study. Finaly, a total of 107 subjects entered our research. PIVKA-Ⅱ was quantitatively measured by Chemiluminescence methods. Kaplan-Meier survival analysis and the Cox proportional hazards model were used to analyze the factors which can affect patient's post surgery survival. RESULTS: Kaplan-Meier survival curve analysis showed that the 1-,2-,3-year survival rate of the patients with high PIVKA-Ⅱ level were 56%, 28% and 16%, respectively, much poor than that of patients with low PIVKA-Ⅱ level, the 1-,2-,3-year survival rate for the latter were 73%, 54% and 46%, respectively(P = 0.002). The 1-,2-,3-year survival rates for patients with high AFP levels were 58%, 35% and 25%, respectively, and the the 1-,2-,3-year survival rates for patients with low AFP levels were 83%, 63% and 51%, respectively(P = 0.006). Multivariate factor results showed that high PIVKA-Ⅱ levels, high AFP levels, the presence of portal vein tumor thrombus and incomplete capsular were independent risk factors for prognosis,HR(95% CI) were 1.99(1.15-3.45), 2.03(110-3.76), 2.72(1.53-4.83) and 2.07(1.07-4.00), respectively. In addition, the prognosis would got worse with the superposition of four independent risk factors. CONCLUSION: Preoperative PIVKA-Ⅱ and AFP levels are associated with HCC patients' s poor post surgery prognosis.

Accordion operation for the bone transport in treating tibial bone defect / 中国骨伤

<p><b>OBJECTIVE</b>To discuss clinical outcomes of accordion operation for the Ilizarov technique in treating tibial bone defects.</p><p><b>METHODS</b>From January 2014 to June 2016, 22 patients with tibial bone defects were treated by Ilizarov bone-transport technique, including 19 males and 3 females with an average age of 44.04 years old ranging from 23 to 60 years old;the length of the bone defects before the bone transport was 5 to 11 cm with an average 7.68 cm; Cause of injury invlved traffic accidents in 14 cases, fall injury in 3, smashing injury in 4, high drop injury in 1; 6 cases were on the left and 16 cases were on the right. The patients were divided into two groups: 11 cases in accordion group were treated by "accordion operation" after bone transport was completed;11 cases in control group were treated by the external fixator locked waiting for bone consolidation after bone transport was completed. All patients were followed up for 18 to 36 months with an average time of 27.9 months. There was no statistical significance between two groups, such as sex, age, length of bone defect(>0.05). Analysis of healing time, healing index and other indicators, and Paley's criterion was used to evaluate the healing effect of bone healing and function recovery of the limb.</p><p><b>RESULTS</b>The result of X-ray evaluation was all patients achieved bone healing. In accordion group, the bone healing time was (365±91) days, the bone healing index was (46.2±3.5) d/cm; in control group, the bone healing time was(435±108) days, the bone healing index was (57.8±3.5) d/cm. There was no statistical significance in the bone healing time between the two groups(=1.648, =0.115);There was statistical significance in the bone healing index between the two groups(=7.754, =0.000). At the final follow-up, according to Paley's criterion, the result in accordion group was excellent in 9 cases, good in 2 cases; in control group, excellent in 8 cases, good in 3 cases. Score was not statistically significant(=-0.479, =0.619). Complications involved nail infection (9 cases in accordion group, 10 cases in control group);local traction pain (2 cases in accordion group, 1 case in control group); axial malalignment>10°(4 cases in accordion group, 3 cases in control group);location difference of the junction of bone defects (3 cases in accordion group, 2 cases in control group);Complications were not statistically significant(>0.05).</p><p><b>CONCLUSIONS</b>Accordion operation for the Ilizarov technique in treating tibial bone defects can shorten the treatment time and consolidation time, and improve the healing index.</p>

Meta-analysis of Yiqi Huoxue method in treating idiopathic membranous nephropathy / 中国中药杂志

The incidence of idiopathic membranous nephropathy (IMN) is increasing year by year, and the clinical research on Chinese medicine treatment of INM is also growing. This study aims to evaluate the efficiency and safety of Yiqi Huoxue method for IMN. Data sources used were from PubMed, EMbase, the Cochrane Library, CBM, CNKI, Wanfang and VIP database. Two researchers independently screened the literature and extracted data. RevMan 5.3 software was used for Meta analysis, and the evidences were graded for the outcomes according to GRADE system by using GRADEprofiler 3.6. Eventually, eleven trials (725 participants) were included in the Meta-analyses. There was statistically significant difference between Yiqi Huoxue method and controls when combining all trials in 24 h UTP [RR=-1.23, 95%CI=(-1.94,-0.53), P=0.000 6] or when combining all trials in ALB [RR=3.56, 95%CI=(1.64, 5.47), P=0.000 3]. Meanwhile, there was statistically significant difference between Yiqi Huoxue method and controls when combining all trials in TC [RR=-0.39, 95%CI=(-0.57, -0.20), P<0.000 1] or when combining all trials in TG [RR=-0.49, 95%CI=(-0.82, -0.15), P=0.004]. However, there was no statistically significant difference between Yiqi Huoxue method and controls when combining all trials in Scr [RR=-3.25, 95%CI=(-9.35, 2.84), P=0.30] or when combining all trials in BUN [RR=-0.81,95%CI=(-2.29, 0.66), P=0.28]. In the statistics, the most frequently used Chinese medicines in clinical application were Astragali Radix, Angelicae Sinensis Radix, Chuanxiong Rhizoma, Codonopsis Radix, Atractylodis Macrocephalae Rhizome and Salvia Miltiorrhiza. The present evidences suggested that Yiqi Huoxue method should be thought highly of in the treatment of IMN, and at the same time, the rational use of traditional Chinese medicine, such as Astragali Radix, Angelicae Sinensis Radix, Chuanxiong Rhizoma also should be paid attention to. However, due to the GRADE evidence grading of the primary outcome measure of 24 h UTP had very low quality, this review can not provide high-quality evidence to prove the clinical efficacy of this method. More well-designed and large-scale multi-center randomized controlled trials should be conducted in the future for verification.

Ginsenoside Rb1 affects the proliferation and osteogenic differentiation of human adipose-derived stem cells in vitro**★ / 中国组织工程研究

BACKGROUND:Various factors can affect the osteogenic differentiation of human adipose-derived stem cel s, and the osteoinductive factor of traditional Chinese medicine is very important for the research of human adipose-derived stem cel s. OBJECTIVE:To investagate the effects of ginsenoside Rb1 on the proliferation and osteogenic differentiation of human adipose-derived stem cel s in vitro. METHODS:The human adipose-derived stem cel s were isolated and cultured in vitro. After passaqed to the third generation, human adipose-derived stem cel s at 2×103/wel were incubated in a 96-wel plate, and treated with 200μL of 0.5, 1.0, 2.0, 4.0,6.0μmol/L ginsenoside Rb1 medium. The human adipose-derived stem cel s in the control group were treated with an equal volume of Dulbecco’s modified Eagle medium. Growth curves were examined by 2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide T colorimetric assay. Alkaline phosphatase activity and osteocalcin content were detected by alkaline phosphatase kit and radio-immunity method, respectively. Calcified nodules were observed using alizarin red O staining. RESULTS AND CONCLUSION:The proliferation viability of human adipose-derived stem cel s was significantly increased after cultured with 0.5μmol/L ginsenoside Rb1. With the increasing of the concentration of ginsenoside Rb1, the mitogenic activity of the cel s was decreased. The 6.0μmol/L ginsenoside Rb1 showed a depressant effect on proliferation. Ginsenoside Rb1 could promote alkaline phosphatase activity and osteocalcin expression in human adipose-derived stem cel s and showed a dose-dependent manner. Calcified nodule formation induced by 4.6 and 6.0μmol/L ginsenoside Rb1 were better when compared with 0.5, 1.0 and 2.0μmol/L ginsenoside Rb1. Ginsenoside Rb1 can promote the proliferation of human adipose-derived stem cel s cultured in vitro in a certain concentration, and in the high concentration, the ginsenoside Rb1 can promote the osteogenic differentiation of human adipose-derived stem cel s. So ginsenoside Rb1 can be used as an osteoinductive factor.

Effects of enhancer of zeste homolog (EZH2) downregulation on the proliferation and invasion of nasopharyngeal carcinoma cell and the possible mechanisms / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of the enhancer of zeste homolog 2 (EZH2) gene on cell growth and invasion of the nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Recombinant lentivirus vector for shRNA delivery of EZH2 was constructed and transfected into 293FT cells. After collecting the viral particles, the NPC cell line 5-8F cells were transfected. The effects of EZH2 silence on cell proliferation and cell cycle were detected using MTT assay, plate colony formation assay and flow cytometry. The migration and invasion of 5-8F cells were determined by wound healing assay and matrigel invasion assay, respectively. The expressions of EZH2 and epithelial-mesenchymal transition (EMT)-related markers at mRNA and protein levels were examined by real-time PCR and Western blot respectively.</p><p><b>RESULTS</b>The expressions of EZH2 mRNA and protein in the transfected 5-8F cells were obviously reduced. MTT assay showed that EZH2 downregulation significantly inhibited the growth of 5-8F/shEZH2 cells (P < 0.001). Colony formation rate (84.44%) of 5-8F/shEZH2 cells was lower than control (31.56%, P = 0.001). Cell cycle analysis showed that most 5-8F/shEZH2 cells were arrested in G0/G1 phase, with a very low ratio of cells in S phase. Wound healing assay indicated that the migration ability of cells silencing EZH2 decreased significantly, and the 48-hour relative migration distance of 5-8F/ShEZH2 cells and control cells was 0.58 ± 0.05, and 0.81 ± 0.02, respectively (P < 0.000). Matrigel invasion assay, showed the invasive capacity of cells silencing EZH2 was significantly inhibited, with less penetrating cells (72.23 ± 4.08) compared to control (150.95 ± 16.27), P < 0.000. The mRNA expressions of epithelial markers E-cadherin and Keratin 18 in the cells silencing EZH2 increased by 177% and 158% respectively, and the mRNA expressions of mesenchymal markers β-catenin and N-cadherin decreased by 18.04% and 41.18% respectively. Similar results also were obtained with Western blot analysis.</p><p><b>CONCLUSION</b>EZH2 significantly enhanced the proliferation and invasion of nasopharyngeal carcinoma cells in vitro, which might be mediated by inducing EMT.</p>

Effect of Epstein-Barr virus nuclear antigen 1 on cell proliferation and cell cycle in nasopharyngeal carcinoma cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the effect of Epstein-Barr virus nuclear antigen 1 (EBNA1) on cell proliferation and cell cycle in nasopharyngeal carcinoma (NPC) cells.</p><p><b>METHODS</b>Recombinant lentivirus that encoded EBNA1 short hairpin RNA (shRNA) was prepared. The C666-EBNA1 (CE) cells were transduced with lentivirus and selected by fluorescence activated cell sorting (FACS) to repress EBNA1 expression. The protein expression levels of EBNA1 were examined by Western blot. The effect of EBNA1 silence on cell proliferation was analyzed by MTT assay and cell growth assay, respectively. Cell cycle was assessed by flow cytometry. The mRNA and protein levels of cell cycle regulators were examined by real-time PCR and Western blot.</p><p><b>RESULTS</b>Recombinant lentivirus encoded EBNA1 shRNA was successfully constructed. The EBNA1 expression in CE cells was significantly reduced by lentivirus-mediated RNA interference. The results of cell counting and MTT assay showed that EBNA1 down-regulation significantly inhibited cell growth in CE-shRNA EBNA1 cells (P < 0.05). Compared with the control group, the percentage of cells in G0-G1 phase was increased from (62.43 ± 6.62)% to (89.66 ± 0.64)% (t = -7.091, P = 0.002), and that in S phase was decreased from (34.93 ± 7.36)% to (7.82 ± 2.44)% (t = 6.095, P = 0.004). The mRNA expressions of c-myc, CDK4, CDK6 and pRb were decreased by 65.60%, 34.06%, 41.05% and 55.29% respectively with the similar results in protein expression levels.</p><p><b>CONCLUSIONS</b>Suppression of EBNA1 may inhibit the growth of nasopharyngeal carcinoma cells in vitro and induce a G1-phase cell cycle arrest, which might be mediated by down-regulation of c-myc, CDK4, CDK6 and pRb.</p>

Transplantation of adipose tissue-derived stem cells promotes soft tissue wound repair in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the effect of local and intravenous transplantation of adipose tissue-derived stem cells (ADSCs) in promoting soft tissue wound healing in rats.</p><p><b>METHODS</b>ADSCs isolated from the adipose tissues of SD rats were cultured in vitro, and the third-passage cells were identified for their capacity of multipotent differentiation. Eighteen SD rats with 1.8 cm² dorsal full-thickness soft tissue defects (0.5 cm deep) were randomized into 3 groups to receive injection of 3.0×10⁶ DiI-labeled ADSCs via the tail vein, local injection of the cells at the wound site, or injection of saline (control). The wound healing was evaluated on days 3, 7, 11, and 14 postoperatively. On day 24 after the injury, tissue samples at the wound site were collected for fluorescent microscopy and HE staining.</p><p><b>RESULTS</b>The ADSCs obtained were capable of adipogenic, osteogenic, and neurogenic differentiation in vitro. ADSCs transplantation significantly promoted wound healing as compared to the control group. Obvious wound contracture was observed in the local injection group on day 3 and in the intravenous injection group on day 7. Fluorescence microscopy revealed DiI-positive cells in the healing wound, and HE staining showed a greater tissue thickness at the wound in the two ADSCs transplantation groups. Compared to the control group, the two ADSCs transplantation groups showed more gland-like structures and better neovascularization at the wound.</p><p><b>CONCLUSION</b>ADSCs can significantly promote wound healing in rats, and local injection of ADSCs allows more rapid and obvious wound healing than tail veil injection of the stem cells.</p>

Development of rapid detection of infectious hypodermal and hematopoietic necrosis virus by loop-mediated isothermal amplification / 病毒学报

Loop-mediated isothermal amplification (LAMP) assay is a novel method of gene amplification with high specificity, sensitivity and rapidity, which can be applied for disease diagnosis in shrimp aquaculture. The method is performed under isothermal conditions with a set of four specially designed primers that recognize six distinct sequences of the target. In the present study, according to the conservative regions of non-structural protein gene NS1, a set of four specific primers were designed, and a rapid detection of IHHNV was established by LAMP assay. The parameters of reaction time and temperature were optimized, and its specificity and sensitivity were assessed. The reactions were carried out at 60 degrees C, 62 degrees C, 63 degrees C, 64 degrees C, 65 degrees C, 66 degrees C, 67 degrees C, 68 degrees C for different time (0 min; 15 min; 30 min; 45 min; 60 min; 75 min). A plasmid pMDIHHNV carrying target sequence of LAMP detection was constructed. Ten-fold serially diluted pMDIHHNV (10(7)-10(0)copies/microL) was used as template for LAMP assay to investigate the detection limit. To determine the specificity, LAMP assays were carried out with DNA templates from other pathogens (White spot syndrome virus; WSSV, Taura Syndrome Virus; TSV, Aeromonas. hydrophila, V. alginolyticus, Vibrio. parahaemolytious, Escherichia. coli). The results showed the optimized LAMP assay for the rapid detection of IHHNV was performed at 65 degrees C for 60 min. The LAMP assay had an unequivocal detection limit of 100 copies/microL, and it was 1,000 times lower than that of PCR. The nucleic acids of other pathogens were not amplified by this LAMP system with the specific primers, which showed a good specificity. The resulting amplicons were detected using visual observation after the addition of SYBR Green I and gel electrophoresis. We investigated the efficacy of UNG (uracil-N-glycosylase) and dUTP in avoiding carry-over contamination in the LAMP assay procedure and explored its effect on the amplification efficiency. Products of LAMP with dUTP adding could be lysed by UNG to avoid LAMP products carry-over contamination effectively. The LAMP assay could be finished within an hour, requiring only a regular laboratory water bath or heat block for reaction and the result could easily be detected using visual observation. Clinically suspected IHHNV-infected shrimp samples were detected by both LAMP and PCR assay, and the result indicated that IHHNV was detected rapidly by LAMP instead of by PCR.

Migration of intravenously injected adipose tissue-derived stem cells in SD rats with soft tissue wound / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the migration of intravenously injected adipose tissue-derived stem cells (ADSCs) in SD rats with soft tissue wound.</p><p><b>METHODS</b>1.8 cm x 1.8 cm full thickness skin defect and 0.5 cm in depth soft tissue defects were made on the back of 6 SD rats. ADSCs were isolated and cultured in vitro for 3 passages. 2.4 x 10(6) ADSCs were labeled with DiI and transplanted into the SD rats through tail vein. Normal skin and wound tissue samples were collected for fluorescent distribution observation 24, 48 days after injection, respectively.</p><p><b>RESULTS</b>Compared to normal skin, more fluorescent positive cells were detected in the margin and deep layer of the wound 24 days after operation. But it is accumulated within dermis and adenoid 48 days after operation.</p><p><b>CONCLUSION</b>Wound can probably induce the migration and accumulation of intravenously transplanted ADSCs.</p>

Association of CXCR4 and SDF-1alpha with organ-specific metastasis of nasopharyngeal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the role of CXCR4/SDF-1alpha axis in organ-specific metastasis of nasopharyngeal carcinoma (NPC) by assessment of CXCR4 expression in NPC cells and SDF-1alpha expression in distant target organs of NPC.</p><p><b>METHODS</b>Thirty patients with NPC and fifteen normal subjects were recruited in this study. The expressions of CXCR4 in NPC and normal cases were identified by RT-PCR and immunohistochemistry (IHC), then the relationship between CXCR4 expression and clinicopathological factors was analyzed. IHC was also used to analyze the SDF-1alpha protein expression in normal cervical lymph nodes (including normal superior and inferior deep cervical lymph nodes), bone marrow, lung, liver, kidney and colon tissues of NPC patients (5 cases/each group).</p><p><b>RESULTS</b>The relative expression level of CXCR4 mRNA in NPC (0.71 +/- 0.22) was significantly higher than that of normal nasopharynx tissues (0.14 +/- 0.07, F = 27.94, P < 0.05). The relative expression level of CXCR4 protein in NPC (1.58 +/- 0.59) was significantly higher than that of normal nasopharynx tissues (0.51 +/- 0.22, F = 17.75, P < 0.05). The high expression levels of CXCR4 mRNA and protein in NPC were closely related to clinical stage, cervical lymph node metastasis and cancer cell differentiation (P < 0.05). SDF-1alpha protein was strongly expressed in normal superior deep cervical lymph nodes, bone marrow, lung and liver (2.35 +/- 0.67), but absent or very poor expression in inferior deep cervical lymph nodes, kidney and colon tissues (0.68 +/- 0.23), and the differences between them were statistically significant (t = 10.13, P < 0.01).</p><p><b>CONCLUSION</b>CXCR4 is closely correlated to metastasis of nasopharyngeal carcinoma. CXCR4/SDF-1alpha axis may play an important role in organ-specific metastasis of NPC.</p>

Effect of Chinese herbal drugs and tetrandrine on TGF-beta1 signaling pathway / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the molecule mechanism of the anti-fibrotic effects of Chinese herbal drugs (Qidan granules) in rats.</p><p><b>METHODS</b>The male rats were randomly divided into four experimental groups: normal group, model group, Qidan group, tetrandrine group. Every group had 10 rats. Normal group were treated with physiologic saline while others were treated with silicon dioxide (50 mg/rat) by intratracheal instillation to induce silicosis. On 30th day Qidan group and Tetrandrine group were treated with Qidan granules (3125 mg/kg) or treated with tetrandrine (22 mg/kg) respectively. All the rats were scarified after 5 months. Lung/body coefficient was measured. Content of hydroxyproline was measured by alkaline hydrolysis. The gene expression of transforming growth factor-beta1 in bronchoalveolar lavage fluid was examined by using enzyme-linked immunosorbent assay (ELISA). The gene expressions of transforming growth factor-beta1, transcription factor Smad 3 and Smad 7 in lung were analyzed by using immunohistochemical technique (SP) and the image analysis.</p><p><b>RESULTS</b>Model group mainly had Grade III approximately IV silicotic nodule while Qidan group and tetrandrine group had Grade II silicotic nodule. In Qidan group and tetrandrine group, lung/body coefficient and content of hydroxyproline and expression of transforming growth factor-beta1 and Smad3 in lung and expression of transforming growth factor-beta1 in bronchoalveolar lavage fluid were lower than those in model group (P < 0.05). Expression of Smad 7 in lung was higher than model group (P < 0.05). Injury of kidney occurred in tetrandrine group.</p><p><b>CONCLUSION</b>Qidan granules and tetrandrine could inhibit expression of both Smad 7 and transforming growth factor-beta1 and promote expression of Smad 3. Qidan granules and tetrandrine could inhibit remarkably silicotic fibrosis in rats. Qidan granules are safer than tetrandrine.</p>