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Objective:To investigate the effect of long non-coding RNA 068 (lncRNA 068) on the migration of a melanoma cell line A375, and to explore its mechanism of action.Methods:From December 2015 to November 2020, 21 patients with pathologically confirmed cutaneous melanoma were collected from Department of Dermatology, Affiliated Hospital of Nantong University, and quantitative PCR (qPCR) was performed to determine the expression of lncRNA 068 in melanoma and paracancerous tissues. LncRNA 068 was overexpressed or knocked down via lentiviral transfection in A375 human melanoma cells in the following experiments. Specifically, A375 cells were divided into lentiviral vector (LV) -green fluorescent protein (GFP) group and LV-lncRNA 068 group to be transfected with a GFP-expressing LV and a LV containing lncRNA 068 respectively in the overexpression experiment, and were divided into LV-LacZ short hairpin RNA (shRNA) group and LV-lncRNA 068 shRNA group to be transfected with a LV containing the reporter gene LacZ-specific shRNA and a LV containing the lncRNA 068-targeting shRNA respectively in the low-expression experiment, with the LV-GFP group and LV-LacZ shRNA group serving as the control groups. Transwell and scratch assays were performed to evaluate cell migration, EdU cell proliferation assay and cell counting kit-8 (CCK8) assay to determine the proportion of proliferative cells and cell viability respectively, and immunofluorescence staining was conducted to evaluate epithelial-mesenchymal transformation in the above groups. Lentivirus-transfected A375 cells from the above groups were inoculated into the axillae of BALB/c nude mice, and tumor volume was measured and calculated every 3 days. After 30 days, all mice were sacrificed, and tumor tissues were resected to measure the tumor volume and weight. Cultured B16F10 cells were subcutaneously inoculated into the back and foot of BALB/c nude mice to construct mouse models of subcutaneously transplanted B16F10 melanoma. After 2 weeks, the mice were sacrificed, and qPCR and Western blot analysis were performed to determine the mRNA expression of inflammatory factors in transplanted B16F10 melanoma and paracancerous tissues, and expression of IκB kinase (IKK) /P65 signaling pathway-related proteins, respectively. Comparisons between 2 groups were done by t test, and comparisons of tumor volume and weight at different time points among groups were done by repeated measures analysis of variance. Results:qPCR showed that the relative expression of lncRNA 068 was significantly lower in human melanoma tissues and transplanted B16F10 murine melanoma tissues (0.414 ± 0.109, 0.717 ± 0.041, respectively) than in the corresponding paracancerous tissues (1.050 ± 0.103, 1.011 ± 0.023, t = 19.48, 10.83, respectively, both P < 0.001). Transwell and scratch assays both showed that the cellular migratory ability was significantly lower in the LV-lncRNA 068 group than in the LV-GFP group (both P < 0.01), and significantly higher in the LV-lncRNA 068 shRNA group than in the LV-LacZ shRNA group (both P < 0.05). Immunofluorescence assay showed significantly increased fluorescence intensity of E-cadherin and decreased fluorescence intensity of N-cadherin in the LV-lncRNA 068 group compared with the LV-GFP group (both P < 0.001), but significantly decreased fluorescence intensity of E-cadherin and increased fluorescence intensity of N-cadherin in the LV-lncRNA 068 shRNA group compared with the LV-LacZ shRNA group (both P < 0.05). In vivo tumor formation experiment in nude mice showed that there were no significant differences in the volume or weight of melanoma between the LV-lncRNA 068 group and LV-GFP group (both P > 0.05), as well as between the LV-lncRNA 068 shRNA group and LV-LacZ shRNA group (both P > 0.05). As qPCR and Western blot analysis showed, the mRNA and protein expression of interleukin-10 (IL-10) and claudin-1 in A375 cells were significantly higher in the LV-lncRNA 068 group than in the LV-GFP group (both P < 0.05), but significantly lower in the LV-lncRNA 068 shRNA group than in the LV-LacZ shRNA group (both P < 0.05). Compared with the paracancerous tissues, B16F10 melanoma tissues showed significantly decreased mRNA expression of IL-10 ( P < 0.01), but significantly increased mRNA expression of IL-6 and tumor necrosis factor-α, as well as protein expression of phosphorylated P65 and phosphorylated IKK ( P < 0.01) . Conclusion:Overexpression of lncRNA 068 can inhibit the migration of A375 melanoma cells, and may affect the development of inflammation and inhibit the epithelial-mesenchymal transformation by inhibiting the IKK/P65 signaling pathway.
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Objective:To investigate the status of work performance among rural doctors in Shanxi province and the factors related to their performance.Methods:From July to September 2019, 528 rural doctors in Shanxi province were selected by multi-stage stratified cluster sampling for the survey. Their job competency, job performance, psychological capital and burnout were evaluated. The structural equation model was used to construct the job performance analysis based on post competency.Results:The total score of post competency of rural doctors in Shanxi province was (188.6±20.3), and the total score of job performance was (71.3±11.5), both of which were in the middle level. The structural equation model was well fitted [goodness-of-fit index (GFI)=0.958, normal fix index (NFI)=0.920, adjusted goodness-of-fit index (AGFI)=0.910, root mean square error of approximation (RMSEA)=0.076]. Post competency, psychological capital and job burnout had direct effects on job performance ( P<0.05), with influence coefficients of 0.64, 0.35 and -0.25, respectively. Job competency indirectly affected job performance through psychological capital and job burnout ( P<0.05). Conclusion:The study suggest that higher post competency would facilitate to improve the work performance of rural doctors.
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The causes of stroke in young people are diverse, most of which are cardiogenic. However, cerebral embolism caused by cardiac myxoma is rare. Intravenous thrombolysis is given in the time window, and the therapeutic effect depends on the nature of embolus. This case is a young female patient, with acute onset, excluding bleeding on the basis of the symptoms, signs and craniocerebral CT. After diagnosed as acute ischemic stroke, the patient was performed an immediate intravenous thrombolysis, though the curative effect was poor. Excluding contraindications six hours after onset, bridging mechanical thrombectomy was performed, postoperative embolus biopsy indicating myxoma. Atrial myxoma was removed in cardiac surgery two months later, and the pathology indicated atrial myxoma. The patient recovered well during the out-of-hospital follow-up, and no further embolization occurred. Therefore, it is suggested that bridging mechanical thrombectomy may be safe and effective for acute ischemic stroke caused by atrial myxoma when intravenous thrombolysis is ineffective, and myxoma resection should be performed immediately after the disease is stable.
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Objective To analyze the inappropriate use of oral drugs in elderly patients admitted in internal medicine, and to guide clinical rational drugs use in elderly patients. Methods One hundred inpatients′ data over 65 years were collected in the department of geriatrics. The potentially inappropriate use of oral drug were analyzed according to the Beers criteria published in 2012 and drugs instructions. Results In 100 patients, (5.81 ± 2.78) kinds of oral drugs had been taken per inpatient. One patient was involved out-of-label usage. There were potentially inappropriate uses of oral drugs in 21 patients. Inappropriate use of oral drugs in three patients were not related to diagnosis or disease, and in 4 patients were related to morbid state, 14 patients used drugs that need to be used with caution in older adults. Conclusions There are off-label uses and potentially inappropriate medications in clinical practice, so clinical doctors should examine and assess the uses of drugs. Clinical doctors should strengthen the understanding and application of the comprehensive geriatric assessment.
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Objective: To investigate the stability of icariin, 2,3,5,4'-tetrahydroxy-stilbene-2-O-β-D-glucoside and hyperoside in Huangmai mixture.Methods: TLC was used to identify astrogaloside IV and Morindae officinalis Radix, and HPLC was used to determine the contents of icariin, 2,3,5,4'-retrahydroxy-stilbene-2-O-β-D-glucoside and hyperoside.The stability of Huangmai mixture under the conditions of accelerated testing and long-term testing was assessed.Results: Astrogaloside IV, Morindae officinalis Radix and icariin showed good stability during the accelerated testing and long-term testing.The contents of 2,3,5,4'-tetrahydroxy-stilbene-2-O-β-D-glucoside and hyperoside declined quickly during the stability testing.Conclusion: Hyperoside and 2,3,5,4'-retrahydroxy-stilbene-2-O-β-D-glucoside are unstable in Huangmai mixture.A novel solid dosage form of Huangmai should be developed to obtain better stability.
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Preparation and in vitro/in vivo evaluation of gestodene [GEST] intravaginal ring [IVR] formulations which can release a constant dose of GEST during 3 weeks were investigated. In present study a reservoir gestodene intravaginal ring, including a gestodene silicone elastomer core and the non-active silicone layer, was reported, which was manufactured by reaction injection moulding at 80[degree]C for 20 min. The raw materials compatibility experiments showed that the silicone elastomer core carrier wouldn't interact with drugs. In vitro release samples were determined by HPLC and the experiment was performed under sink conditions. The equation of cumulative release verse time was Y=64.76x+5.44 [r=0.9998], performing zero-order release at about the target dose of 60 [micro]g/day over 21 days. Drug release increased with temperature elevating from 45 to 55[degree]C, which could be attributed to optimizing the prescription. In addition, the pharmacokinetic and safety studies of gestodene intravaginal ring were evaluated in female New Zealand White rabbits. The GEST in plasma was analyzed by LC-MS/MS and the results proved that the correlation between in vitro and in vivo was relatively well
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Objective To compare the efficacy of intralesional glucocorticoid injection with a needle-free injector versus an ordinary injector for the treatment of keloid. Methods A total of 60 patients with keloid were enrolled and randomly divided into two groups by using a random number table to receive intralesional injection of compound betamethasone with a needle-free injector(n=31)or an ordinary injector(n=29). The injection was given at a dose of 0.2 ml/cm3 once every 3 weeks for 3 sessions. Parameters for therapeutic efficacy were assessed, adverse reactions were recorded, and clinical pictures were taken before and after each treatment. Statistical analysis was carried out by the Mann-Whitney U test and chi-square test with the SPSS 19.0 software. Results Compared with the ordinary injector group, the needle-free injector group showed significantly different injection time during the first and second treatment (U=299.000, 773.500, respectively, both P=0.000), as well as duration of pain after the first injection(U=730.000, P=0.003). After three sessions of treatment, there was a significant difference in the volume, height, hardness of keloid, scores for pain, itching and appearance, and number of injection points between the needle-free injector group and ordinary injector group (U=295.000, 336.500, 264.000, 464.000, 451.500, 308.000, 233.500, P=0.001, 0.007, 0.000, 0.041, 0.043, 0.003, 0.001, respectively). No significant differences were observed in the incidence of adverse reactions between the two groups(all P>0.05). Moreover, the interval for lesion recurrence was significantly shorter in the needle-free injector group(11.8 days, 95%CI:10.96-12.6 days)than in the ordinary injector group(21.2 days, 95%CI:13.96-28.45 days). Conclusion Compared with the ordinary injector, the needle-free injector shows better efficiency for the treatment of keloid with decreased difficulty in injection and improved compliance in patients.
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Objective To investigate the efficacy of dermabrasion combined with aminolevulinic acid-based photodynamic therapy (ALA-PDT) for the treatment of nasal nodular basal cell carcinoma (nBCC).Methods Twentyfive patients who were pathologically diagnosed as nasal nBCC with lesion area ＞ 1 cm2 but no bone or cartilage involvement,were included in this study and treated with dermabrasion combined with ALA-PDT.At first,the part of tumor protruding outside the skin was removed by artherectomy,then dermabrasion was carried out.The wound surface was topically treated with 20％ aminolevulinic acid solution for 3-4 hours away from light immediately after surgery,then irradiated with LED light at a mean dose of 100 J/cm2 for 20 minutes.ALA-PDT was performed once a week for 3 consecutive weeks.The degree of and time required for wound healing were assessed,and tumor recurrence,cicatrization and appearance outcomes were observed during 1 year after surgery.Efficacy was assessed comprehensively.Results No postoperative wound infection occurred in these patients,and the average time for wound healing was (11.2 ± 1.3) days.During 1 year after the treatment,no recurrence was found,while cicatricial contracture developed in 1 case,mild proliferative scar in 3 cases,and depressed scar in 4 cases.All the patients were satisfied with the treatment outcomes,except 1 patient who was basically satisfied.Conclusions Dermabrasion combined with ALA-PDT is easy to operate with rapid wound healing,low postoperative recurrence rate and high degree of patient satisfaction,and is worthy of clinical promotion.
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Objective To study the correlation of blood drug concentration determination of sodium valproate in children with epilepsy by using enzyme-multiplied immunoassay test (EMIT) and high performance liquid chromatography (HPLC), so as to provide basis of selection in clinical determination of sodium valproate ( VPA) .Methods 200 blood samples of epileptic children taking sodium valproate were collected, blood concentration were determined by EMIT and HPLC method, and the correlation was analyzed.Results The following regression equation was obtained by determination results of HPLC (X) and EMIT (Y): Y=1063.517X-331.351(r=0.933), which had significant correlation (P<0.05).The plasma concentration of VPA determined by EMIT was higher than that by HPLC.Conclusion The determination results of VPA concentration in epileptic children by using EMIT and HPLC has significant correlation, and it should choose the suitable method to determine the blood drug concentration of VPA based on the actual situation.
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Objective To identify differentially expressed proteins in keloid tissue and to explore the proteomic characteristics of keloid by utilizing capillary high-performance liquid chromatography (LC) and tandem mass spectrometry (MS).Methods Tissue specimens were obtained from the lesions of 5 patients with non-familial keloid and normal skin of 5 human controls.After extraction and in-solution digestion of proteins,the resultant peptides were separated and identified by LC-MS/MS.Label-free detection and quantitation of peptides were carried out by using the DeCyder MSTM software.Results A total of 570differentially expressed peptides were identified between non-familial keloid and normal tissue (all P ＜ 0.05),which correspond to 1221 proteins.A difference greater than 1.5 folds was observed in the expressions of 293 proteins,including 124 up-regulated and 169 down-regulated proteins in non-familial keloid tissue.Conclusion Differences exist in the expression of proteins between non-familial keloid and normal skin tissue.
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Objective To retrospectively analyze the therapeutic effect of wide surgical excision combined with low-dose adjuvant interferon-alpha 2b on cutaneous malignant melanoma.Methods A total of 32 patients with cutaneous malignant melanoma received wide surgical excision after preoperative examination and staging.The excisions were performed with a margin measuring 1-2 cm from the visible lesions or biopsy scars.Surgical modalities included direct suture after excision(4 patients),dactylolysis or toe amputation(6 patients),free skin grafting(15 patients),random skin flap transfer(3 patients)and pedicle skin flap transfer(4 patients).Lymph nodes were selectively dissected in 9 patients with regional transfer of lymph nodes,and inguinal lymph nodes were cleared away in 2 patients.One week after the operation,patients received adjuvant therapy with subcutaneous injection of interferon-alpha 2b(3 million IU,thrice per week)for one to three years.Results Preoperative tumor staging revealed 21 cases of cutaneous malignant melanoma at stage Ⅱ,and 11 cases at stage Ⅲ.The excisions healed by the first stage in all the patients.Up to June 2011,2 patients had been lost to follow up,5 patients with stage Ⅲ melanoma had died.Survival was observed in all of the 4 patients receiving 1-year follow up,12 of 13 patients receiving 1-3 year follow up,5 of 7 patients receiving 3-5 year follow up,and 4 of 6 receiving 5-year follow up.Of the 25 surviving patients,regional lymph node metastasis was observed in 8 patients,which developed within 2 years after the operation in 2 patients.The adjuvant therapy with interferon-alpha 2b lasted 3 years in 8 patients,and more than 1 year in 11 patients.Side effects were mild.Conclusion Wide surgical excision plus low-dose interferon-alpha 2b is effective for the treatment of stage Ⅱ and Ⅲ cutaneous malignant melanoma with lower local recurrence and higher survival rate.
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<p><b>OBJECTIVE</b>To establish a baking method of Platycladi Cacumen Carbonisatum for providing a new idea to Carbonic Herbs' research.</p><p><b>METHOD</b>Samples were prepared in an oven for different time at different temperatures separately. Then the fingerprints of the samples were determined by UPLC. According to the standard fingerprint, the similarities of the samples' fingerprints were compared.</p><p><b>RESULT</b>The similarities of 3 samples, which were baked at 230 degrees C for 20 min, 30 min and at 240 degrees C for 20 min, were above 0.96.</p><p><b>CONCLUSION</b>According to the similarities of the fingerprints and in view of the appearances, Platycladi Cacumen Carbonizing should be baked at 230 degrees C for 20 min.</p>
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Chromatography, High Pressure Liquid , Cupressaceae , Chemistry , Drugs, Chinese Herbal , Plant Extracts , Technology, Pharmaceutical , MethodsABSTRACT
0.05).One case developed hemorrhage of venous plexus in the posterior pedicle screw fixation group.And complications occurred in 6 cases(4 of superior nerve injury,1 of intra-incision hematoma and 1 of esophageal injury).There were significant differences between 2 groups(P
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[Objective] A retrospective study was performed to evaluate the outcome of three-dimensional correction for adolescent idiopathic scoliosis(AIS)and discuss biomechanics of posterior three-dimensional correction surgery and measures for the prevention of spinal imbalance.[Methods]Thirty-one patients with AIS were classified and treated with three-dimensional correction surgery from 2005 to 2007.There were 18 male and 13 female with the average age of 15.2 years(range,11 to 18 years).The cases were classified according to the Lenke surgical classification,including type I 8 cases,type Ⅱ 4 cases,type Ⅲ 8 cases,type Ⅳ 2 case,type Ⅴ 5 cases,type Ⅵ 4 cases.The major curve Cobb's angles,the distance of apical vertebra shift and trunk shift,were measured and analyzed in the reoperative,postoperative and final follow-up radiographies.The average preoperative coronal Cobb's angle was 54?,the distance of both apical vertebra shift and trunk shift was 5.8 cm and 2.6 cm respectively.[Results]The mean time of follow-up was 16 months(range 1 to 3 years).The average postoperative coronal Cobb's angle was 21?,as the correction was 61%,the lost angle at the last follow-up was 4.0?.The postoperative distance of apical vertebra shift was 2.5cm,as the correction was 56%,the lost distance at the last follow-up was 0.5 cm.The distance of trunk shift at the last follow-up was 1.0 cm.[Conclusion]Posterior three-dimensional correction surgery is a good corrective method in the treatment of AIS.The spinal mechanical balance should be noticed during the operaion.
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[Objective]To evaluate the outcome on patients with primary infection of intervertebral space of the thoracolumbar spine following combined one-stage debridement,strut grafting and internal fixation.[Method] From May 1999 to February 2006,18 patients with spontaneous infection of intervertebral space were analyzed retrospectively.These infections occurred at thoracic spine in 3 cases(16.7%),thoracolumbar junction in 2 cases(11.1%),lumbar spine in 10 cases(55.5%),and lumbosacral junction in 3 cases (16.7%).Their clinical data and outcome were studied.None of all had previous spinal surgery or spinal injection,and their diagnosis were confirmed by histopathology after surgery.All patients were treated by one-stage debridement,strut grafting and internal fixation after failure of nonsurgical treatment.Antibiotics was used following debridement in the infection sites,and was continued to be administered after surgery.Patients were evaluated before and after surgery in terms of pain,neurologic level,sagittal spinal balance,and radiologic fusion.[Result]Postoperatively,all patients experienced significant relief of symptoms,and no fever.Six patients showed improved neurological status(at least one grade improvement on Frankel's functional classification).Only 3 of 18 cases had a positive culture result that pathogen identified Staphylococcus aureus.Histological examination of the specimen after surgery revealed that inflammation occurred in all patients,but no evidence of tuberculosis.The period of follow-up ranged 13 to 45 months with a mean of 26 months.No evidence of recurrence or residual infection was observed in any patient.Postoperative radiological evaluation revealed that implants were stable,there was no phenomena of prosthesis subsidence,hook dislodgment and failure restoration of spinal segments height.Solid bony fusion was obtained in all patients.Eight patients were misdiagnosed for spinal tuberculosis.[Conclusion] One-stage eradication of the infection,strut grafting and internal fixation are effective treatments for primary infection of intervertebral space.The presence of the instrumentations at the site of infection has no negative influence on the course of infection healing.
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AIM:To develop a high performance liquid chromatography -mass spectrometry method for the determination of ?-Lipoic acid in plasma,to study the pharmacokinetics of ?-Lipoic acid and evaluate the bioequivalence of two preparations in healthy subjects. METHODS:A single oral dose of 200 mg ?-Lipoic acid capsules and tablets was given to 22 healthy male volunteers according to an open randomized 2 way crossover design. Plasma concentrations of ?-Lipoic acid were determined by HPLC-MS method. The pharmacokinetic parameters and relative bioavailability were calculated to evaluate the bioequivalence of the two preparations. RESULTS:The pharmacokinetic parameters of the two products were as follows:tmax were (16?4) and (20?20) min,?max were (1490?359) and (1537?290) ng/mL,AUC0-t were (59559?18456) and (58210?15080) ng?mL-1?min,AUC0-∞ were (60068?18556) and (58634?15126) ng?mL-1?min,respectively. The relative bioavailability of AUC0-t and AUC0-∞ were (102.97?18.28)% and (103.09?18.26)%. CONCLUSION:The result demonstrated that two formations are bioequivalent by statistical analysis of variant,two one-side t-test and 90% confidential interval.
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AIM:To establish the fingerprint for Cacumen Platycladi (carbonized) by HPLC. METHODS:The column of Lichrospher C 18 (250 mm?4.6 mm 5 ?m)was used. The mobile phase consisted of 0.5‰ trifluoroactic acid-methanol with gradient elution. The detective wavelength was at 375 nm,and the flow rate was 1.0 mL/min. Different habitats were compared by Similarity Evaluation System for Chromatographic Fingerprint of CMM Version 2004A. RESULTS:The fingerprint consisted of 14 common peaks. The mutual mode of HPLC fingerprints was set up and the similarity of the crude drugs was in the range of 0.178-0.963. The standard HPLC fingerprint of Cacumen Platycladi (carbonized) was established too. CONCLUSION:This method is accurate and reliable and provides a scientific basis for the quality control of Cacumen Platycladi (carbonized).