Objective:To prepare poly-L-lactic acid(PLLA)electrospun nanofibers carrying icariin(ICA)(ICA /PLLA)and to evaluate the effects of the ICA /PLLA on MC3T3-E1 cells.Methods:ICA solution was dispersed into PLLA solution,and electrospun fibers were fabricated by W/O emulsion method.The morphology of ICA /PLLA was observed by SEM.The in vitro release kinetics of ICA /PLLA was examined.The attachment of MC3T3-E1 cells on ICA /PLLA was examined by propidiumiodide(PI)labling and ob-served under fluorescent microscope.The proliferation of the cells was measured by MTT assay.The differentiation of the cells was ob-served by alkaline phosphatase (ALP)assay.Results:In vitro,ICA was effectively released from ICA /PLLA for 22 days,cells were attached well on the surface in all groups,ICA did not affect the proliferation of MC3T3-E1 cells(P >0.05),but increased the ALP activity(P <0.05)of the cells.Conclusion:ICA /PLLA can effectively control the release of ICA and promote the differentiation of MC3T3-E1 cells.

Objective:To construct a recombinant adenovirus containing cytosine deaminase( CD ) gene and thymidine kinase( TK ) fusion gene for the gene therapy research of malignant tumors. Methods: A recombinant cosmid containing CD and TK fusion gene was constructed, and then mixed with DNA TPC and co transfected to the 293 cells by calcium phosphate coprecipitation. Results: The results of restriction and PCR showed the insertion was right and the recombinant adenovirus generated contained the CD and TK fusion gene without replication competent adenovirus. Conclusion: The recombinant adenovirus generated is E1 and E3 deleted and contains double suicide gene needed, which can be further studied for gene therapy.

Objective To observe the changes of esophageal motility in patients with achalasia (AC) before and after peroral endoscopic myotomy (POEM ) and to evaluate the effects of POEM on esophageal motility in AC .Methods A total of 35 patients with AC received POEM .The esophageal motility in response to different food swallows (5 mL liquid and 2 cm × 2 cm × 2 cm solid food) was evaluated by high-resolution manometry (HRM) system before operation and one month after operation .The changes of parameters of esophageal body and lower esophageal sphincter (LES) were analyzed and compared before and after operation .The t-test ,non-parametric test or single factor analysis of variance was performed for statistical analysis .Results Before POEM operation ,lower esophageal sphincter pressure (LESP) of 35 patients in response to liquid swallows and solid swallows was (28 .94 ± 18 .70) mmHg (1 mmHg=0 .133 kPa) and (26 .41 ± 11 .57) mmHg ,respectively ;after operation it was (16 .02 ± 5 .46) mmHg and (15 .82 ± 5 .04) mmHg ,respectively ;and the differences were statistically significant (t= 4 .338 and 4 .726 ,both P<0 .01) .Before operation ,4 s integrated relax pressure (4 s IRP) during liquid swallows and solid swallows was (27 .18 ± 14 .63) mmHg and (28 .46 ± 11 .15) mmHg ,respectively ;after operation it was (12 .22 ± 6 .75) mmHg and (14 .54 ± 7 .83) mmHg ,respectively ;and the differences were statistically significant (t= 5 .902 and 5 .436 ,both P< 0 .01) .And after operation intra boluspressure (IBP) of liquid swallows and solid swallows also decreased compared to that before operation (t=5 .075 and 2 .944 ,both P< 0 .01) .Lower esophageal sphincter relaxation rate (LESRR) during liquid swallows and solid swallows increased compared to that before operation (t= -2 .990 , P< 0 .01 ;t=-0 .340 ,P>0 .05) .There was no difference in the distal contractile integral (DCI) and distal esophageal peristaltic amplitude in subtype Ⅰ and Ⅲ patients before and after operation (all P>0 .05 ) .All those parameters decreased in subtype Ⅱ patients (Z= -2 .704 and -2 .489 ,P< 0 .05 ;Z= -1 .929 and-0 .747 , P> 0 .05 ) . Proximal esophageal peristalsis was observed in two patients after operation , however there was no integrated esophageal body peristalsis .Clinical symptoms quickly relieved in all patients after POEM operation and clinical score significantly decreased compared to that before operation (0 .86 ± 1 .19 vs 8 .16 ± 1 .84 ,t=20 .605 , P<0 .05) .Conclusions POEM can effectively relieve LES relaxation disorder in AC patients and improve esophageal body peristalsis to a certain degree .The efficacy is regardless of AC types ,and further studies are need to shed light on the long-term efficacy .The long-term efficacy still need further follow-up study .

Objective To evaluate the effect of video- assisted thoracoscopic pulmonary segmentectomy in patients with isolated pulmonary arteriovenous fistula (PAVF). Methods A retrospective analysis was performed on 10 patients with PAVF in the department of thoracic surgery of the first affiliated hospital of Nanjing Medical University between January 2010 and December 2016. Computed tomography angiography (CTA) and three-dimensional reconstruction were performed before operation, and all patients accepted video-assisted thoracoscopic pulmonary segmentectomy. Results The diagnosis of PAVF was identified by CTA, with maximum diameter of tumor of 3.0- 5.0 cm. No perioperative mortality or postoperative complications were observed including bleeding, hemoptysis, serious air leakage, and bronchopleural fistula. The lesions were completely removed in all 10 patients, and no patients converted to open surgery intraoperatively. Blood gas analysis showed that oxygen partial pressure before operation, in the first day after operation and the third month after operation was (62.5 ± 6.7), (70.2 ± 4.8) and (75.4 ± 4.8) mmHg (1 mmHg = 0.133kPa) respectively; which was significantly increased successively (P<0.05). After a follow-up time of 3-30 months, no recurrences were observed. Conclusions Video- assisted thoracoscopic pulmonary segmentectomy guided by preoperative CTA and three-dimensional reconstruction is a very effective method for the treatment of isolated PAVF.

Objective To investigate the clinicopathological characteristics and postoperative prognosis analysis of intra-thoracic localized Castleman disease (LCD).Methods The clinical data of 9 patients with intra-thoracic LCD who accepted surgical treatment were retrospectively analyzed.There were 5 males and 4 females,with age of (32.8 ± 10.9) years.Two patients complained of chest pain,1 patient suffered from paraneoplastic pemphigus,and the rest were diagnosed by physical examination.Four cases were diagnosed with LCD by preoperative CT examination.Results All patients underwent surgical resection.Four patients were performed open surgery and 5 patients had video assisted thoracic surgery.All patients accepted radical surgery.But 2 of these patients had postoperative complications.One patient was the injury of phrenic nerve and another was pericardial effusion.Patho-histological showed hyaline vascular type of Catleman disease in all patients.All patients survived without recurrence during the follow-up for 2-53 months.Conclusions Intra-thoracic is rare and liable to misdiagnosed.For increasing the preoperative diagnosis rate of LCD,the combined application of imaging tests is important,and clinicians and radiologists should also enhance the awareness of this disease.Complete surgical resection of the tumor is the best therapeutic alternative for intra-thoracic LCD.

ObjectiveTo investigate the effect of heat treatment combined with narrow band ultraviolet B(NB-UVB) on cultured normal human melanocytes in vitro.MethodsMelanocytes were isolated from the foreskin of normal human,cullured in vitro,and irradiated with NB-UVB of different doses(20,30,50,70,90,120 and 180 mJ/cm2).Then,MTT assay was performed to evaluate the proliferation and activity of melanocytes to determine the optimal dose of UVB for the next experiment.Melanocytes were classified into 3 groups to be treated with heat at 42 ℃ for 1 hour (heat group),irradiated with UVB at 50 mJ/cm2 (UVB group),or irradiated with UVB at 50 mJ/cm2 followed by heat treatment at 42 ℃ for 1 hour (combination group),daily for 3 successive days; those receiving no treatment served as the control.After 24-hour culture following the last treatment,tyrosinase activity was evaluated with L-dopa as the substrate,melanin content was detected by NaOH assay,and cell cycle stages were determined by flow cytometry.ResultsNB-UVB irradiation decreased the viability of melanocytes in a dose-dependent manner,and the optimum dose of UVB was 50 mJ/cm2.The tyrosinase activity of melanocytes was 0.244 ± 0.018 and 0.310 ± 0.015 respectively in the UVB group and combination group,and increased by 3.8％ (P ＜ 0.05) and 31.9％ (P ＜ 0.05) respectively compared with the control group (0.235 ± 0.018); the melanin content was 0.201 ± 0.016 and 0.286 ± 0.019,respectively in the UVB group and combination group,and increased by 17.5％ (P ＜ 0.05 ) and 67.3％ (P ＜ 0.05) compared with the control group (0.171 ± 0.016).In comparison with the control group,the percentage of melanocytes in G1 phase was decreased by 23.94％ in the UVB group(P＜ 0.05) and 33.51％ in the combination group(P ＜ 0.05),while that in S phase and G2 phase increased by 15.35％ (P ＜ 0.05 ) and 11.93％ (P ＜ 0.05),respectively in the UVB group,and 17.76％ (P ＞ 0.05) and 16.08％ (P ＞ 0.05),respectively in the heat group.ConclusionHeat treatment and NB-UVB can synergistically enhance the tyrosinase activity and accelerate melanogenesis,proliferation and differentiation,of melanocytes.

ObjectiveTo explore the effects of heat treatment and ultraviolet B (UVB) radiation alone or in combination on the expression of heat shock protein (HSP) 72 in human epidermal melanocytes.Methods Melanocytes were obtained from human foreskin,and subjected to primary culture.After 3 to 5 passages,the melanocytes were classified into 4 groups:control group (receiving no treatment),heat treatment group (treated with heat at 42 ℃ for 1 hour every day for 3 days),UVB group(irradiated with UVB at 50 mJ/cm2 daily for 3days),combination group(treated with heat at 42 ℃ for 1 hour followed by irradiation with UVB at 50 mJ/cm2daily for 3 days).After another 2- to 6-hour culture following the last treatment,melanocytes were collected and subjected to real time PCR and Western blot for the detection of HSP72 mRNA and protein expression,respectively.ResultsThe mRNA and protein expressions of HSP72 were significantly higher in the heat treatment group and combination group than in the control group (mRNA:6.584 ± 0.871 and 7.269 ± 0.454 vs.0.975 ± 0.089,both P ＜ 0.001; protein:2.022 ± 0.058 and 2.080 ± 0.045 vs.0.532 ± 0.033,both P ＜ 0.001 ),but was similar between the UVB group and control group (mRNA:0.832 ± 0.084 vs.0.975 ± 0.089,P ＞ 0.05;protein:0.546±0.021 vs.0.532 ± 0.033,P ＞ 0.05).The ANOVA of factorial design showed that neither heat treatment nor UVB irradiation had interaction effect on the mRNA or protein expression of HSP72 (F =2.106,1.399 respectively,both P ＜ 0.05).ConclusionsHeat treatment can cause an increase in the expression of HSP72,which may enhance the function of melanocytes and protect melanocytes from UVB induced damage.

ObjectiveTo observe the increasing effect of infrared irradiation on tyrosinase activity and melanin content in cultured normal human epidermal melanocytes in vitro and to explore the optimal dose of infrared irradiation.MethodsEpidermal melanocytes were isolated from normal human foreskin tissue,and subjected to primary culture.Methyl thiazolyl tetrazolium(MTT) assay was performed to evaluate the effect of different doses(0,20,60,80,100,140,240,320 J/cm2)of infrared light on the proliferation of melanocytes and to select the optimal irradiation dose.Then,melanocytes were irradiated with infrared light at the optimal dose for 3 consecutive days followed by the determination of tyrosinase activity,melanin content,and cell cycle via dopa oxidation assay,NaOH solubilization method and flow cytometry,respectively.ResultsThe best intervention dose of infrared light was 80 J/cm2.The tyrosinase activity(A492 nm) and melanin content(A492 nm)were 0.3601 ± 0.0301 and 0.2748 ± 0.0243 respectively in melanocytes after irradiation with infrared light of 80 J/cm2 for 3 days,significantly higher than those in unirradiated melanocytes(0.3114 ± 0.0341,0.2325 ±0.0254,respectively,both P ＜ 0.05),with an increase rate of 15.64％ and 18.19％ respectively.Cell cycle analysis revealed a decline in cell percentage in G1 phase(P ＜ 0.01 ) but a concomitant increase in cell percentage in G2 and S phase (both P ＜ 0.05) in irradiated melanocytes compared with unirradiated melanocytes.ConclusionsThe optimal dose of infrared light is 80 J/cm2 for the irradiation of melanocytes,and this dose of infrared light can increase melanin content,tyrosinase activity,differentiation and proliferation of melanocytes.

OBJECTIVETo observe the efficacy and safety of Qizhi Jiangtang Capsule (QJC) in treating stage 3b diabetic kidney disease (DKD) patients with macroalbuminuria.

METHODSPatients who conformed to the diagnostic criteria of stage 3b DKD were randomly assigned to two groups according to random digital table, the experiment group and the control group, 84 in each group. All patients received a two-week elution period, and then were treated with basic Western therapy. Patients in the experiment group took QJC, 5 pills per time, 3 times a day, while those in the control group took Valsartan Capsule 160 mg each time, once daily. The observation period of follow-ups was limited within 6 months, and the time points were set as the baseline, 1st month, 3rd month, and 6th month. Systolic blood pressure (SBP), diastolic blood pressure (DBS), 24 h urine protein quantitative (24 h UPQ), plasma albumin (ALB), and serum creatinine (SCr) were detected and recorded, and estimated glomerular filtration rate (eGFR) was calculated. The occurrence of hypoglycemic reaction, coagulation disorder, gastrointestinal tract reaction, allergy, hyperkalemia, doubling of creatinine, and overall adverse events were observed and recorded at same time.

RESULTSFinally 81 patients in the experiment group and 80 patients in the control group were effectively included. Compared with the baseline level, SBP and DBS obviously decreased in the control group at month 1 of treatment (P < 0.05), and more significantly decreased at month 6 of treatment (P < 0.01). SBP at month 1, 3, and 6 of follow-ups; DBS at month 6 of follow-ups was lower in the control group than in the experiment group (P < 0.05). At month 1, 3, and 6 of follow-ups, 24 h UPQ of the experiment group was significantly lower than the baseline level (P < 0.01). It was also significantly lower than the level of the control group at the same time point (P < 0.05). There was no significant difference in 24 h UPQ at month 1, 3, and 6 of follow-ups between the control group and the baseline level (P > 0.05). ALB of the experiment group showed an increasing trend. It was significantly higher than the baseline level at month 6 (P < 0.05), which was also higher than that of the control group at same period (P < 0.05). There was no significant difference in the ALB level in the control group (P > 0.05). SCr of two groups showed an increasing trend. SCr of the experiment group was significantly higher at month 1, 3, and 6 follow-ups than the baseline level (P < 0.05). But the increment of SCr was higher in the control group than in the experimental group, and obviously higher than the baseline levels (P < 0.05). eGFR of both groups showed a decreasing trend. The decrement was higher in the control group than in the experimental group (P < 0.05). The proportion of progression of renal functions at month 1, 3, and 6 of follow-ups in the experimental group was 0.0% (0 case), 9.55% (8 cases), and 21.4% (18 cases), while they were 8.3% (7 cases), 21.4% (18 cases), and 40.5% (34 cases) in the control group. There was no statistical difference in the proportion of progression of renal functions between the two groups at month 3 and 6 of follow-ups (P < 0.05). There was no statistical difference in the incidence of adverse reactions between two groups (P > 0.05).

CONCLUSIONQJC could effectively reduce urinary protein of patients with stage 3b DKD, and delay the progression of renal functions.

Adult ; Albumins ; analysis ; Albuminuria ; drug therapy ; Blood Pressure ; drug effects ; Creatinine ; blood ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerular Filtration Rate ; Humans ; Male ; Middle Aged ; Tetrazoles ; therapeutic use ; Treatment Outcome ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan

Objective To analyze the correlation between the dry eye symptom and signs in middle-aged and aged people by Lipiview(R) ocular surface interferometry.Methods A total of 170 eyes from 85 middle-aged and aged people (55-80 years old) were examined with Standard Patient Evaluaion of Eye Dryness(SPEED) questionnaire and Lipiview(R) ocular surface interferometry,meanwhile obtaining the consent of the subjects.Then the correlation between the symptom and Lipid Layer Thinckness (LLT) and partial blink rate were analyzed.Results SPEED questionnaire score was 6.13 ±4.60.LLT was(74.88 ± 21.16) nm.LLT variability was 4.47 ± 3.40.The partial blink rate was 0.62 ± 0.36.The Spearman correlation analysis indicated that the nega-tive correlation was shown between the SPEED questionnaire score and LLT (r =-0.823,P =0.000),SPEED questionnaire score and LLT variability(r =-0.268,P =0.018).The positive correlation were found between LLT and Std.Dev of LLT (r =0.339,P =0.030).However,the results of the SPEED score were uncorrelated with partial blink rate (P ＞ 0.05).Conclusion The results from this study demonstrate that correlation among symptom,LLT and LLT variability,but poor correlation between symptom and partial blink rate.