Comparison of Exosomes Derived from Fibroblasts and Mesenchymal Stem Cells for the Treatment of Acute Wound Healing / 中山大学学报(医学科学版)

ObjectiveTo compare the therapeutical effect of exosomes derived from fibroblasts and mesenchymal stem cells on acute wound healing. MethodsPrimary human dermal fibroblasts （hDF） were isolated， cultured and identified. Human bone marrow mesenchymal stem cell exosomes （hMSC-EXO） and hDF exosomes （hDF-EXO） were extracted by ultracentrifuga tion. After 24 h of coincubation with hDF-EXO or hMSC-EXO， hDFs proliferation and migratory capacity were evaluated by cell counting kit-8 （CCK8） assay and scratch test. Full-thickness cutaneous wounds were created on 8-week-old female C57BL/6 mice， and topically applied with PBS （control）， hDF-EXO or hMSC-EXO. Wounds were measured at day 0， 2， 4， 7， and the uptake of exosomes in wound was observed at day 1. Quantitative PCR （qPCR） analysis was performed to detect the mRNA expression levels of TNF-α， IL-6， IL-1β， IL-10 in wound at day 1. HE staining was conducted to analyze the histological structure of wounds at day 7， while immunofluorescence staining was used to examine expression of PDGFR-α、α-SMA、Ki67. ResultshDF exhibited certain fibrolast-like characteristics with respect to expression of cell surface markers and specific proteins. hDF-EXO and hMSC-EXO presented exosomal morphology， size， and markers， and both concentrations were not statistically different （P>0.05）； CCK8 assay showed that both exosomes promoted hDF cell viability， compared with the negative control （P<0.01）， and hDF-EXO group had greater cell viability than hMSC-EXO group （P<0.01）. Scratch test indicated that hDF-EXO induced a significant increase in scratch healing rate versus the negative control （P<0.01）， hMSC-EXO （P<0.05）. In vivo experiments showed wound tissues took up exosomes at day 1. qPCR detected TNF-α， IL-6， IL-1β expression levels in wound at day 1 were lower in exosomes group than in the control group， and were the lowest in hMSC-EXO group （all P<0.01）. Wound areas were measured smaller at day 7 in exosomes group than in the control group （all P<0.01） and hDF-EXO group had better closure than hMSC-EXO group （P<0.05）. HE staining revealed that compared with control group， scar， incomplete epidermis and few collagen deposition remained in the hMSC-EXO group， whereas hDF-EXO group showed re-epithelialization， continuous neo-epidermis and regenerated dermis. Immunofluorescence staining suggested that the number of fibroblasts， myofibroblasts， proliferating cells was higher in both exosomes group than that in the control group， especially the highest in hDF-EXO group. ConclusionOur study shows both exosomes accelerate wound healing， whereas hDF-EXO is more effective in promoting fibroblasts proliferation， migration， transition to myofibroblasts， and hMSC-EXO may play a role in inhibiting inflammatory reaction during early stage of wound healing.

Ershiwuwei Shanhu Pills regulate Akt/mTOR/GSK-3β signaling pathway to alleviate Alzheimer's disease mice / 中国中药杂志

The present study investigated the mechanism of the Tibetan patent medicine Ershiwuwei Shanhu Pills(ESP) in alleviating Alzheimer's disease in mice via Akt/mTOR/GSK-3β signaling pathway. BALB/c mice were randomly assigned into a blank control group, a model group, low(200 mg·kg~(-1)), medium(400 mg·kg~(-1)) and high(800 mg·kg~(-1)) dose groups of ESP, and donepezil hydrochloride group. Except the blank control group, the other groups were given 20 mg·kg~(-1) aluminum chloride by gavage and 120 mg·kg~(-1) D-galactose by intraperitoneal injection for 56 days to establish Alzheimer's disease model. Morris water maze was used to detect the learning and memory ability of mice. The level of p-tau protein in mouse hippocampus and the levels of superoxide dismutase(SOD), malondialdehyde(MDA), catalase(CAT), and total antioxidant capacity(T-AOC) in hippocampus and serum were detected. Hematoxylin-eosin staining and Nissl staining were performed for the pathological observation of whole brain in mice. TdT-mediated dUTP nick-end labeling(TUNEL) staining was employed for the observation of apoptosis in mouse cortex. Western blot was adopted to detect the protein levels of p-mTOR, p-Akt, and GSK-3β in the hippocampus. Compared with the model group, the ESP groups showcased alleviated pathological damage of the whole brain, decreased TUNEL positive cells, reduced level of p-tau protein in hippocampus, and risen SOD, CAT, and T-AOC levels and declined MDA level in hippocampus and serum. Furthermore, the ESP groups had up-regulated protein levels of p-mTOR and p-Akt while down-regulated protein level of GSK-3β in hippocampus. Therefore, ESP can alleviate the learning and memory decline and oxidative damage in mice with Alzheimer's disease induced by D-galactose combined with aluminum chloride, which may be related to Akt/mTOR/GSK-3β signaling pathway.

Prognostic and survival analyses of early-stage diffuse large B-cell lymphoma of Waldeyer′s ring / 中华放射肿瘤学杂志

Objective@#To analyze the clinical efficacy and prognostic factors of patients with early-stage diffuse large B-cell lymphoma of Waldeyer′s ring (WR-DLBCL) treated with CHOP-based chemotherapy.@*Methods@#A total of 137 patients diagnosed with WR-DLBCL admitted to our hospitalfrom 2006 to 2018 were enrolled, including 22 patients with stage Ⅰ and 115 patients with stage Ⅱ WR-DLBCL. All patients received CHOP-based chemotherapy, of whom 62 receiving rituximab and 87 receiving involved-field radiotherapy. The overall survival (OS), progression-free survival (PFS) and local recurrence-free survival (LRRFS) were calculated by Kaplan-Meier method. Log-rank test, was conducted for univariate analysis and Cox’s regression model was performed for multivariate analysis.@*Results@#The 5-year OS, PFS, and LRFFS in the whole group were 78.6%, 69.5% and 83.2%, and 87.5%, 80.2%, 90.9% in the comprehensive treatment group, and 64.2%, 53.6% 72.9% in the chemotherapy group, respectively. Univariate analysis showed that lactate dehydrogenase (LDH), international prognostic index score, large mass, rituximab, chemotherapy cycle and comprehensive treatment were the prognostic factors of OS and PFS. In addition, LDH, large mass and comprehensive treatment were the prognostic factors associated with LRFFS. Multivariate analysis demonstrated that LDH, comprehensive treatment mode and rituximab were the prognostic factors of OS. LDH and comprehensive treatment mode were the prognostic factors associated with PFS. LDH was a prognostic factor of LRFFS.@*Conclusion@#Patients with early-stage WR-BLBCL obtain excellent clinical prognosis. In the era of rituximab treatment, chemotherapy combined with radiotherapy remains an efficacious treatment of early-stage WR-BLBCL.

Clinical evaluation of cervicectomy in the treatment of cervical intraepithelial neoplasia with cervicovaginal shortening / 中国实用妇科与产科杂志

OBJECTIVE: To investigate the clinical efficacy of cervicectomy for cervical intraepithelial neoplasia(CIN)in patients with cervicovaginal shortening.METHODS: Retrospectively analyze the clinical data of the 120 cases of CIN treated in Shengjing Hospital of China Medical University from April 2014 to November 2018.Cervicectomy was performed because of cervicovaginal shortening caused by menopausal or peri-menopausal cervical atrophy or cervical surgery.The clinical treatment,efficacy and prognosis of the patients were reviewed.RESULTS: The mean age of the 120 patients was 55.2 years(range:35-77 years).The indications of operation included:persistent abnormal cervical cytology test(7),CIN2(42),CIN3(70),squamous carcinoma of the cervix(1);peri-menopausal and menopausal patients with obvious cervicovaginal atrophy(100),premenopausal patients with natural short cervix(2),and obvious cervicovaginal shortening caused by cervical surgery(18).The mean operating time was 23.2 min(range 10-30 min),the mean bleeding volume was 7.8 mL(range:5-20 mL),and the mean height of cervix resected was 2.59 cm(range:2-3 cm).No secondary injury,bleeding or other postoperative complications occurred during surgery;cervical postoperative wounds healed well;only one case developed cervical adhesion after surgery.The postoperative histologic diagnosis were compared with the preoperative histologic diagnosis,in which 45 degraded(37.50%),42 consistent(35.00%),and 33 upgraded(27.50%).HPV conversion rate 3 months after cervicectomy was 80.81%(80/99),and total HPV conversion rate was 88.89%(88/99).A total of 29 patients underwent secondary surgery,23 underwent total hysterectomy,and 6 underwent extensive hysterectomy and pelvic lymphadenectomy.All patients were followed up,once every 3 to 6 months,and median follow-up time was 29.5 months(range 4-59 months).All patients recovered well after surgery;only 2 cases showed positive margins,and only 2 cases of residual disease and 1 case of recurrence were found during follow-up.CONCLUSION: For patients with cervical intraepithelial neoplasia of cervicovaginal shortening,cervicectomy is a safe,effective and relatively microinvasive treatment.

Effect analysis of topical fluoride varnish on caries prevention of first permanent molars in school-age children / 口腔疾病防治

Objective @#To investigate the effectiveness of fluoride varnish in preventing first permanent molars caries inschool-age children and provide necessary information for oral health instruction. @*Methods@#378 children aged 6-7 years old in the first grade of primary school in dahua country, hechi city, Guangxi province (1 053 first permanent molars) were randomly recruited into 2 groups. The test group (542 first permanent molars in 191 children) was treated with fluoride varnish one time in a half year and given oral hygiene instruction 3 times; The control group (511 first permanent molars in 187 children) was only given oral hygiene instruction 3 times. All first permanent molars were checked after 18 months later for caries. @*Results@#After intervention, the caries prevalence rate in the test group raised from 21.99% to 27.75% compared with the control group from 24.60% to 37.43%. The caries incidence rate in test and control group was 5.76% and 12.83% respectively. There was a significant difference between 2 groups regardless of caries prevalence, caries incidence, the DMFT reduction, the DMFs reduction, the occlusal caries prevalence, the pit and fissure caries prevalence, the proximal caries prevalence. @*Conclusion @#Semi-annual application of fluoride varnish is effective in the prevention of dental caries in first permanent molars , which was worth of being popularized in clinic.

Effects of Tongxinluo Capsule on Platelet Activating Factor, Vascular Endothelial Function, Blood Flow of Thrombolysis in Myocardial Infarction in Acute Myocardial Infarction Patients after Delayed Percutaneous Coronary Intervention / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore effects of Tongxinluo Capsule (TC) on platelet activating factor (PAF), vascular endothelial function, thrombolysis in myocardial infarction (TIMI) blood flow, and heart function in acute myocardial infarction (AMI) patients after delayed percutaneous coronary intervention (PCI).</p><p><b>METHODS</b>Totally 80 AMI inpatients were recruited at Department of Cardiology, People's Hospital of Jiangxi Province, from Jan. 2008 to Sep.2013. Those in line with inclusion criteria were randomly assigned to TC treatment group and the conventional treatment group by random digit table, 40 in each group. Besides, another 40 healthy subjects from examinees at Outpatient Department were recruited as a healthy control group. PCI was performed after 1-week treatment. Then blood samples were collected, and then blood contents of CD62P, CD63, GP II b/III a, ET-1, NO, and plasma von Willebrand factor (vWF) levels were detected. Coronary TIMI blood flow and corrected TIMI frame count (CTFC) were determined during PCI. Meanwhile, noninvasive blood pressure (BP) and heart rate (HR) were recorded before and after PCI, and cardiac function measured. They were compared with the healty control group.</p><p><b>RESULTS</b>Compared with the healthy control group, blood contents of CD62p, CD63, GP II b/IIIa receptor compound, vWF, and ET-1 significantly increased, but NO significantly decreased in AMI patients (all P < 0.05). After 1-week intervention of TC, blood contents of CD62p, CD63, GP II b/IIIa receptor compound, vWF, NO, and ET-1 significantly decreased (P < 0.05, P < 0.01). Compared with the conventional treatment group at the same time point, blood contents of CD62p, CD63, GP II b/IIIa receptor compound, vWF, and ET-1 decreased more significantly in the TC group (P < 0.05, P < 0.01), increased NO levels were also more obviously seen (P < 0.01). The aforesaid parameters changed more obviously at day 30, as compared with those changes at week 1 (P < 0.05, P < 0.01). The TIMI blood flow grade and CTFC were more obviously improved after PCI in the two treatment groups. Better TIMI blood flow was seen in the TC group. TIMI level 3 blood flow rate was higher in the TC group than in the conventional treatment group with statistical difference (P < 0.05). The left ventricular ejective factor (LVEF) after PCI was obviously elevated in the TC group and the conventional treatment group (P < 0.01), and the improvement was more obviously seen in the TC group (P < 0.05). There were 6 cases of recurrent angina, 3 cases of ventricular tachycardial (VT)/ventricular fibrillation (VF), 6 cases of heart failure (HF), 1 case of cardiac sudden death in the conventional treatment group, with the total incidence of cardiovascular events being 40% (16/40). There were 2 cases of recurrent angina, 2 cases of VT/VF, 2 cases of HF, no cardiac sudden death in the TC treatment group, with the total incidence of cardiovascular events being 15% (6/40). There was statistical difference in the recurrent rate of cardiovascular events between the two groups (χ² = 2.27, P < 0.05).</p><p><b>CONCLUSION</b>TC not only could prevent coronary embolism of AMI patients after delayed PCI, attenuate vascular endothelial injury, but also could improve TIMI blood flow, and strengthen cardiac systolic function.</p>

Application of percutaneous balloon mitral valvuloplasty in patients of rheumatic heart disease mitral stenosis combined with tricuspid regurgitation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Tricuspid regurgitation (TR) is frequently associated with severe mitral stenosis (MS), the importance of significant TR was often neglected. However, TR influences the outcome of patients. The aim of this study was to investigate the efficacy and safety of percutaneous balloon mitral valvuloplasty (PBMV) procedure in rheumatic heart disease patients with mitral valve (MV) stenosis and tricuspid valve regurgitation.</p><p><b>METHODS</b>Two hundred and twenty patients were enrolled in this study due to rheumatic heart disease with MS combined with TR. Mitral balloon catheter made in China was used to expand MV. The following parameters were measured before and after PBMV: MV area (MVA), TR area (TRA), atrial pressure and diameter, and pulmonary artery pressure (PAP). The patients were followed for 6 months to 9 years.</p><p><b>RESULTS</b>After PBMV, the MVAs increased significantly (1.7 ± 0.3 cm 2 vs. 0.9 ± 0.3 cm 2 , P < 0.01); TRA significantly decreased (6.3 ± 1.7 cm 2 vs. 14.2 ± 6.5 cm 2 , P < 0.01), right atrial area (RAA) decreased significantly (21.5 ± 4.5 cm 2 vs. 25.4 ± 4.3 cm 2 , P < 0.05), TRA/RAA (%) decreased significantly (29.3 ± 3.2% vs. 44.2 ± 3.6%, P < 0.01). TR velocity (TRV) and TR continue time (TRT) as well as TRV × TRT decreased significantly (183.4 ± 9.4 cm/s vs. 254.5 ± 10.7 cm/s, P < 0.01; 185.7 ± 13.6 ms vs. 238.6 ± 11.3 ms, P < 0.01; 34.2 ± 5.6 cm vs. 60.7 ± 8.5 cm, P < 0.01, respectively). The postoperative left atrial diameter (LAD) significantly reduced (41.3 ± 6.2 mm vs. 49.8 ± 6.8 mm, P < 0.01) and the postoperative right atrial diameter (RAD) significantly reduced (28.7 ± 5.6 mm vs. 46.5 ± 6.3 mm, P < 0.01); the postoperative left atrium pressure significantly reduced (15.6 ± 6.1 mmHg vs. 26.5 ± 6.6 mmHg, P < 0.01), the postoperative right atrial pressure decreased significantly (13.2 ± 2.4 mmHg vs. 18.5 ± 4.3 mmHg, P < 0.01). The pulmonary arterial pressure decreased significantly after PBMV (48.2 ± 10.3 mmHg vs. 60.6 ± 15.5 mmHg, P < 0.01). The symptom of chest tightness and short of breath obviously alleviated. All cases followed-up for 6 months to 9 years (average 75 ± 32 months), 2 patients with severe regurgitation died (1 case of massive cerebral infarction, and 1 case of heart failure after 6 years and 8 years, respectively), 2 cases lost access. At the end of follow-up, MVA has been reduced compared with the postoperative (1.4 ± 0.4 cm 2 vs. 1.7 ± 0.3 cm 2 , P < 0.05); LAD slightly increased compared with the postoperative (45.2 ± 5.7 mm vs. 41.4 ± 6.3 mm, P < 0.05), RAD slightly also increased compared with the postoperative (36.1 ± 6.3 mm vs. 28.6 ± 5.5 mm, P < 0.05), but did not recover to the preoperative level. TRA slightly increased compared with the postoperative, but the difference was not statistically significant (P > 0.05). The PAP and left ventricular ejection fraction appeared no statistical difference compared with the postoperative (P > 0.05), the remaining patients without serious complications.</p><p><b>CONCLUSIONS</b>PBMV is a safe and effective procedure for MS combined with TR in patients of rheumatic heart disease. It can alleviate the symptoms and reduce the size of TR. It can also improve the quality-of-life and prognosis. Its recent and mid-term efficacy is certain. While its long-term efficacy remains to be observed.</p>

Effect of pomegranate peel polyphenol gel on cutaneous wound healing in alloxan-induced diabetic rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pomegranate (punica granatum) belongs to the family Punicaceae, and its peel has been used as a traditional Chinese medicine because of its efficacy in restraining intestine, promoting hemostasis, and killing parasites. Pomegranate peel has been reported to possess wound-healing properties which are mainly attributed to its polyphenol extracts. The purpose of this study was to investigate the effect of pomegranate peel polyphenols (PPP) gel on cutaneous wound healing in diabetic rats.</p><p><b>METHODS</b>Alloxan-induced diabetic rats were given incisional wounds on each side of the mid-back and then treated daily with PPP gel (polyphenol mass fraction = 30%) post-wounding. Rats were sacrificed on days 4, 7, 14, and 21 post-wounding to assess the rates of wound closure, histological characteristics; and to detect the contents of hydroxyproline, production of nitric oxide (NO), and activities of NO synthase (NOS), as well as the expressions of transforming growth factor-β1 (TGF-β1), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF) in wound tissue.</p><p><b>RESULTS</b>Wound closure was significantly shortened when PPP gel was applied to the wounds of diabetic rats. Histological examination showed the ability of PPP gel to increase fibroblast infiltration, collagen regeneration, vascularization, and epithelialization in the wound area of diabetic rats. In addition, PPP gel-treated diabetic rats showed increased contents of hydroxyproline, production of NO, and activities of NOS and increased expressions of TGF-β1, VEGF, and EGF in wound tissues.</p><p><b>CONCLUSION</b>PPP gel may be a beneficial method for treating wound disorders associated with diabetes.</p>

Development of ELISAs for the detection of urogenital chlamydia trachomatis infection targeting the pORF5 protein / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To prepare antibodies against pORF5 plasmid protein of Chlamydia trachomatis and develop double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) for the detection of genital C. trachomatis infections.</p><p><b>METHODS</b>The pORF5 protein was expressed in Escherichia coli and used to immunize BALB/c mice and New Zealand rabbits to produce monoclonal antibodies (mAbs) and polyclonal antibody (pAb) for DAS-ELISAs. Clinical samples from 186 urogenital infection patients (groups I) and 62 healthy donors (groups II) were detected in parallel by the DAS-ELISAs developed in this study and by IDEIA PCE commercial ELISA.</p><p><b>RESULTS</b>Two hybridoma cell lines, named 2H4 and 4E6, stably secreting specific mAbs against pORF5 were obtained. The mAb 2H4 was recognized by 32 (17.20%, positive recognition rate) and 25 (13.44%), mAb 2H4 by 0 (0%) and 2 (3.22%) samples from groups I and II, respectively. The sensitivities of mAbs 2H4 and 4E6 were 92.11% and 77.78% and the specificities were 100% and 96.88%, respectively in relation to the IDEIA PCE commercial ELISA. The sensitivities of detection for the DAS-ELISAs were 10 ng/mL (based on 2H4) and 18 ng/mL (based on 4E6).</p><p><b>CONCLUSION</b>Two DAS-ELISAs were developed in this study that provided a feasible and effective assay that could be considered alternative tools for the serodiagnosis of C. trachomatis infection.</p>

Predictive value of three preoperative nutritional assessment methods in postoperative complications of gastrointestinal cancers / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To compare the value of three preoperative nutritional assessment methods, European nutrition risk screening 2002(NRS 2002), mini-nutrition assessment(MNA) and subjective global assessment(SGA), in predicting postoperative complications of gastrointestinal cancer patients.</p><p><b>METHODS</b>A total of 235 patients with gastrointestinal cancers, including 31 esophageal cancers, 82 gastric cancers, and 122 colorectal cancers, in our hospital from January 2012 to June 2013 were prospectively enrolled. Preoperative nutritional status was evaluated with above 3 methods respectively. Postoperative complication rates were compared among different preoperative nutritional status.</p><p><b>RESULTS</b>According to SGA score, the morbidity of severe-moderate, mild and no malnourished patients was 40.5%(17/42), 25.3%(22/87) and 14.2%(15/106) respectively(P<0.01). According to MNA score, the morbidity of patients with malnutrition, at risk of malnutrition and without malnutrition was 32.9%(23/70), 24.7%(18/73) and 14.1%(13/92) respectively(P<0.05). According to NRS 2002, the morbidity of patients at malnutrition risk and without malnutrition risk was 27.6%(27/98) and 19.7%(27/137) respectively(P>0.05). Multiple regression analysis revealed that both SGA and MNA scores were predictive factors for the development of postoperative complications(both P<0.01). The sensitivity of SGA score for predicting complications was higher compared to MNA score (90.7% vs. 79.6%), while the specificity was similar(49.7% vs. 50.8%).</p><p><b>CONCLUSIONS</b>Both SGA and MNA scores can effectively predict the development of postoperative complications in gastrointestinal cancer patients, and SGA score has better sensitivity. SGA score is recommended for decision-making regarding preoperative nutrition support.</p>

Proteolysis induce factor in the digestive systematic cancer cachexia patients: its expression and role in cancer cachexia / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To demonstrate the expression of proteolysis induce factor(PIF) in the gastrointestinal(GI) cancer cachexia patients and evaluate its role in cancer cachexia.</p><p><b>METHODS</b>Examination of PIF was performed in urine samples from 28 GI cancer cachexia patients, 13 GI cancer patients without cachexia, and 12 weight loss patients with benign disease. PIF was added to the mice cultured C2C12 muscle cells, then the protein kinase B(Akt) phosphorylation and morphological change were measured.</p><p><b>RESULTS</b>The positive rate of PIF in urine of 28 cancer cachexia patients was 53.6%(15/28). In the other two groups, no positive result was detected. PIF could successfully induce Akt phosphorylation, cell atrophy, metamorphosis, and death. The peak of this phosphorylation could be detected after half an hour of the initiation of PIF at a concentration of 4 nmol/L.</p><p><b>CONCLUSIONS</b>PIF is specifically and highly expressed in GI cancer cachexia patients' urine. PIF can induce cancer cachexia possibly by activating Akt phosphorylation and inducing downstream proteolysis.</p>

Application of PHILOS plate through mini-open deltoid-splitting approach for the treatment of proximal humeral fractures / 中国骨伤

<p><b>OBJECTIVE</b>To study clinical effects of PHILOS (proximal humeral internal locking system) plates through mini-open deltoid-splitting approach for the treatment of proximal humeral fractures.</p><p><b>METHODS</b>From March 2006 to August 2010, 22 patients with proximal humeral fractures were treated with PHILOS plates through mini-open deltoid-splitting approach. According to Neer classification, 6 cases were type II, 15 cases were type III and 1 case was type IV. Through the anterolateral approach to the shoulder, anterolateral vertical incision of 4 cm length was perforrmed from 1 cm under acromion, and separated deltoideus muscle vertically to touch the fracture,reduced the end of fracture directly and indirectly. PHILOS plate was inserted downward into anterolateral surface of humerus through deltoideus muscle, the distal end and proximal end was fixed by locking screws. The Neer score for shoulder function was evaluated within 1 year after operation.</p><p><b>RESULTS</b>The operative time ranged from 30 to 70 minutes with an average of 45 minutes. No blood transfusion was required during the operation, and all incisions healed in stage I. All the patients were followed up, and the duration ranged from 6 to 18 months with a mean time of 12.5 months. All the fractures healed up perfectly, and the union time ranged from 6 to 12 weeks. According to Neer criteria for shoulder joint function, 10 patients got an excellent result, 9 good, 2 poor and 1 bad. There were no complications such as axillary nerve injuries, screw loosening, steel plate breakage, dislocation of shoulder joint and necrosis of humeral bone.</p><p><b>CONCLUSION</b>PHILOS plate through mini-open deltoid-splittin approach for the treatment of proximal humeral fractures has follow advantages: simple recover,minor-injuries and small tissue invasion, which is an ideal method to treat proximal humeral fractures.</p>

Cloning and cellular localization of pORF8 plasmid protein of Chlamydia trachomatis / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone the plasmid protein pORF8 of Chlamydia trachomatis and localize its expression in Chlamydia-infected cells.</p><p><b>METHODS</b>pORF8 gene was amplified and cloned into pGEX-6p vector, and the pORF8 fusion protein was expressed in E.coli XL1 Blue. After purification with glutathione-conjugated agarose beads, the pORF8 fusion protein was used to immunize BALB/c mice to generate polyclonal antibodies against pORF8 protein. The antibodies obtained were used to localize the plasmid protein pORF8 in Chlamydia-infected cells with immunofluorescence assay (IFA).</p><p><b>RESULTS</b>The pORF8 gene 744 bp in length was successfully cloned and the GST fusion protein with a relative molecular mass of 54 000 was obtained. The cellular distribution pattern of the plasmid protein pORF8 was similar to that of the major outer membrane protein (MOMP), a known C. trachomatis inclusion body protein, but not to that of chlamydial protease-like activity factor (CPAF, a secreted protein).</p><p><b>CONCLUSION</b>The plasmid protein pORF8 is localized on the bacterial organism as an inclusion body protein in C. trachomatis-infected cells. The cellular location of pORF8 protein can potentially provide important insights into the pathogenesis of C. trachomatis.</p>

Functional reconstruction of ischemic contracture in the lower limb / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To discuss the method of functional reconstruction of ischemic contracture in the lower limb and propose a classification protocol for ischemic contracture in the lower limb based on its severity and prognosis.</p><p><b>METHODS</b>A total of 42 patients with ischemic contracture in the lower limb were included in this study. According to different types of disturbance and degrees of severity, surgical reconstructions consisting of nerve decompression, tendon lengthening or transfer, intrinsic foot muscle release and sural-tibial nerve anastomosis were performed in every patient.</p><p><b>RESULTS</b>Postoperatively, all patients were able to walk on flat ground. Drop foot was corrected in 10 patients, and 5 patients still felt some difficulty during stair activity. Split Achilles tendon transfer to flexor hallucis longus tendon was performed in 12 patients, and their walking stability was improved. Seven patients accepted ipsilateral sural-tibial nerve anastomosis, and sensitivity recovery reached to S2 in 2 patients and S3 in 5 patients.</p><p><b>CONCLUSIONS</b>Ischemic contracture in the lower limb is a devastating complication after lower limb trauma. The prevention of contracture is much more important than the treatment of an established contracture. Split Achilles tendon transfer to flexor hallucis longus tendon and sural-tibial nerve anastomosis, which was initially implemented by us, could improve the functional recovery of ischemic contracture in lower limbs, and thus provides a new alternative for functional reconstruction of ischemic contracture in the lower limb.</p>

Effects of microRNA-29 family members on proliferation and invasion of gastric cancer cell lines / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>MicroRNAs have emerged as post-transcriptional regulators that are critically involved in the biologic behavior of cells. This study was designed to investigate the effect of members of the microRNA-29 family on the expression of cell division cycle 42 (Cdc42) and their roles on proliferation, migration, and invasion of gastric cancer cells.</p><p><b>METHODS</b>We detected microRNA-29s and Cdc42 expression in gastric cancer cells by real-time polymerase chain reaction (PCR) and Western blot analysis. Negative controlled RNA (ncontrol), microRNA-29 family members (microRNA-29a, -29b, and -29c), and Cdc42-specific small interfering RNA (si-Cdc42) were chemically synthesized and transfected into SGC7901 and BGC823 gastric cancer cells, which have a relatively low expression of microRNA-29s and a relatively high expression of Cdc42. The expression of Cdc42 and the phosphorylation of its downstream molecular PAK1 expressions were determined by Western bolt analysis. Cell Counting Kit-8 was used to measure cell proliferation, and wound-healing and invasion assays were used to examine the abilities of migration and invasion.</p><p><b>RESULTS</b>Similar to si-Cdc42, the ectopic expression of microRNA-29 family members significantly reduced the expression of Cdc42 and its downstream molecular PAK1 phosphorylation levels. Consistently, ectopic expression of microRNA-29s inhibited proliferation and migration in gastric cancer cells. Invasive cell counts of the SGC7901, ncontrol/SGC7901, si-Cdc42/SGC7901, microRNA-29a/SGC7901, microRNA-29b/SGC7901, and microRNA-29c/SGC7901 cell groups were 84.0+/-4.2, 71.7+/-4.6, 16.3+/-3.2, 15.7+/-3.8, 16.3+/-3.0, and 16.7+/-3.1, respectively. The invasive cell counts of the BGC823, ncontrol/BGC823, si-Cdc42/BGC823, microRNA-29a/BGC823, microRNA-29b/BGC823, and microRNA-29c/BGC823 cell groups were 199.0+/-10.5, 146.3+/-9.7, 72.7+/-8.2, 86.7+/-8.5, 86.0+/-8.5, and 73.3+/-8.3, respectively (P<0.05).</p><p><b>CONCLUSIONS</b>Members of the microRNA-29 family can obviously inhibit cell proliferation, migration, and invasion of gastric cancer cells by targeting Cdc42.</p>

Establishment and biological characteristics of oxaliplatin-resistant human colon cancer cell lines / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Chemotherapy is the main treatment for colon cancer, while multidrug-resistance is the main reason for chemotherapy failure and tumor relapse. This study was to establish two oxaliplatin-resistant colon cancer cell lines and evaluate their biological characteristics.</p><p><b>METHODS</b>Oxaliplatin-resistant colon cancer cell lines SW620/L-OHP and lovo/L-OHP were established in vitro by continuous exposure to oxaliplatin (L-OHP) of low and gradually increased concentration. Growth curve, cross-resistance and resistance index of the oxaliplatin-resistant cell lines to various anti-cancer agents were determined by CCK8 assay. The expressions of P-glycoprotein (P-gp), multidrug-resistance protein 1 (MRP1) and MRP2 were detected by Western blot. Cell cycle distribution as well as the expression of CD133 and CD44 were measured by flow cytometry.</p><p><b>RESULTS</b>It took 10 months to establish the SW620/L-OHP and LoVo/L-OHP cell lines with stable resistance to oxaliplatin. Cross-resistance to 5-fluorouracil, etoposide, cisplatin, vincristine and epirubicin but not to paclitaxel was observed. Longer doubling time, higher proportion of cells in G(0)/G(1) phase and lower proportion in G(2)/M phase were observed in the two oxaliplatin-resistant cell lines compared with their parental cell lines. The expression of MRP2 in the oxaliplatin-resistant cells was up-regulated, while those of P-gp and MRP1 had no significant change. CD133 was overexpressed while CD44 level remained unchanged in SW620/L-OHP and LoVo/L-OHP cells.</p><p><b>CONCLUSIONS</b>SW620/L-OHP and LoVo/L-OHP cell lines show a typical and stably resistant phenotype and may be used as research models.</p>

Treatment of type C3 distal radius fracture resulted from high-energy injuries by volar plate in combination with external fixator / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>With a type C3 distal radius fracture it is extremely difficult to maintain the reduction and to restore congruity of the articular surface because the support for the volar and dorsal bone cortex has been lost. An external fixator crossing wrist in combination with Kirschner wire (K-wire) fixation was popularly used by most studies in recent years. But loss of reduction often occurred especially in the volar sides.</p><p><b>METHODS</b>A total of 30 cases of type C3 distal radius fracture were treated by a volar buttress plate combined with a transarticular external fixator and other techniques such as K-wire fixation and bone grafting if necessary. The postoperative volar tilt angles, ulnar inclinations, radial heights, range of motion, grip strength and complications were recorded to assess the therapeutic effects.</p><p><b>RESULTS</b>Of the 30 patients, 27 patients were followed up for 12 - 29 (mean 18) months and all fractures healed in 8 - 13 weeks after surgery (mean 10 weeks). The wrist function was excellent in 8 cases, good in 16 cases and fair in 3 cases according to the Sarmiento scoring system (modified by Stewart).</p><p><b>CONCLUSIONS</b>For type C3 comminuted fracture with severe volar and dorsal instability, fixation by volar buttress plate combined with transarticular external fixator should be adopted. Dorsal instability could be further stabilized by other techniques such as Kirschner wire fixation, and bone grafting.</p>

Gene profiling of MAPK pathway in human osteosarcoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the functional effects of MAPK pathway in the pathogenesis of human osteosarcoma.</p><p><b>METHODS</b>Gene microarray (Human Genome U133A, Affymetrix) was used to screen the differential expression of genes involved in MAPK pathway between osteosarcoma cell lines and 3 osteoblastic cell lines. KEGG metabolic pathway analysis was performed among significantly increased or decreased genes using the MATLAB software. Immunohistochemical technique was used to detect the expressions of ERK1/2, JNK and p38 proteins among 48 osteosarcoma and benign 24 osteoblastic tumor samples.</p><p><b>RESULTS</b>Using an entrance limit of > or = 2.0, 18 differentially expressed MAPK pathway-related genes were selected (10 up-regulated, 8 down-regulated) to mapped to the MAPK pathway of KEGG which are all important node genes. The positive rates of ERK1/2, JNK and p38 proteins were 83.3% (40/48), 72.9% (35/48) and 85.4% (41/48) in osteosarcomas,and 12.5% (3/24), 8.3% (2/24) and 16.7% (4/24) in the control group, respectively. The positive rates and expression intensities were statistically different between the 2 groups (P<0.01).</p><p><b>CONCLUSION</b>MAPK pathway plays an important role in the pathogenesis of osteosarcoma. ERK, JNK and p38 form an intercoordinating network and regulate the cell proliferation, differentiation, apoptosis, invasion and migration in osteosarcoma.</p>

Rapid detection of norwalk GII with reverse transcription loop-mediated isothermal amplification / 病毒学报

To develop a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Norwalk GII. 4 primers which recognized 6 distinct regions on the RNA-dependent RNA polymerase gene of Norwalk GII were designed and used for LAMP assay. Norwalk GII RNA was amplified under isothermal conditions (65 degrees C) for 120 min, and LAMP results were then judged with naked eye, SYBR Green I staining, electrophoretic analysis and restriction digestion. To evaluate the specificity of the RT-LAMP, 48 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses were tested. To compare the sensitivity of the RT-LAMP with that of conventional RT-PCR, Norwalk GII RNA was serially diluted and amplified by RT-LAMP and RT-PCR, respectively. With 46 fecal specimens of Norwalk GII, observation with naked eyes, SYBR Green I staining and electrophoretic analysis were able to detect the PCR products in the RT-LAMP assay. The specificity of RT-LAMP products was also confirmed by digestion of the RT-LAMP products with restriction enzymes. No RNA amplification was observed in 2 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses. The specificity of the RT-LAMP assay with regard to RT-PCR were 100% for Norwalk GII. The detection limits of RT-LAMP was 15.6 pg/tube for Norwalk GII and similar to that of a RT-PCR assay. Compared to RT-PCR, the RT-LAMP assay has been proven to be a rapid, sensitive, specific and accurate method for detection of the Norwalk GII in fecal specimens, and that RT-LAMP assay is potentially useful for the rapid detection of Norwalk GII from fecal specimens in outbreaks of infectious diarrhea.

Combination of volar buttress plate with external fixator for the distal radial fractures of type C3 caused by high-energy injuries / 中国骨伤

<p><b>OBJECTIVE</b>To preliminarily explore the effect of combination of volar buttress plate with external fixator for the distal radial fractures of type C3 caused by high-energy injuries.</p><p><b>METHODS</b>From January 2001 to June 2007, 13 patients with distal radial fracture of type C3, 9 males and 4 females aged from 26 to 47 (average 37 years), were treated with volar buttress plate combined with external fixator plus the techniques of K-wires and bone grafting as necessary, whose effects were evaluated preliminarily through comparing the volar tilt, radial inclination, radial shortening and wrist function.</p><p><b>RESULTS</b>Followed up from 7 to 29 months (average 18 months), the volar tilt, radial inclination, radial shortening and wrist function of all patients recovered remarkably. Nine patients achieved excellent and 4 good according to Sarmiento score (modified by Stewart) in the radiological manifestation, while 5 patients displayed excellent, 6 good, and 2 fair according to Gartland-Werley functional assessment system.</p><p><b>CONCLUSION</b>1) Volar buttress plate could support the valor cortex in order to prevent comminuted fragment from displacing and maintain volar tilt and to provide the volar fulcrum for external fixator. 2) External fixator, with the assistance of volar fulcrum, could maintain the volar tilt and the height of distal radius and help unload the fossa. 3) Supplemental K-wires fixation and the bone graft may assist fracture stable.</p>