Objective To evaluate the efficacy of laparoscopic Miles surgery at differented periods.Methods Radical excision(Miles) was performed under a laparoscope on 33 patients from January 2002 to August 2002(group 1),and then on 34 cases from July 2006 to May 2007(group 2) in our hospital.Clinicopathological and follow-up data of the two groups were collected and analyzed.Results No significant difference was found in blood loss(50-200 ml vs 40-210 ml,Z=1.477,P=0.787),number of resected lymph nodes(both 3-5,Z=-0.089,P=0.233),gas expel(in 1-3 days after the surgery vs 1-4 days,Z=-1.622,P=0.574),and postoperative hospital stay(4-9 days vs 5-9 days,Z=-1.834,P=0.346) between the two groups.However,the patients in group 1 had significantly longer operation time and higher medical cost than group 2(175-210 min vs 120-150 min,Z=4.238,P=0.026;23400-27500 yuan vs 19863-22744 yuan,Z=5.283,P=0.003).Conclusion After 4-year development,the operation time and cost of laparoscopic Miles surgery has been decreased.

Objective To investigate the genetic polymorphism of DXS8378 STR locus of chromosome X in Chinese Lisu,Pumi and De'ang populations in Yunnan and construct relative standard allelic ladders.Methods After being amplified by PCR,different STR allelic fragments were isolated from the PAG electrophoresis.The STR allelic fragments were extracted by kit and reamplified by PCR to obtain purified allelic fragments.Next,the purified allelic fragments were subcloned individually into the PUC plasmid vectors,and the size and structure of the inserts were confirmed by the analysis of their DNA sequences.Then we transfected it into competent E.coli DH5?TM cells,and finally,the recombinant plasmids DNA with the inserts were used as template for reamplification to generate the standard ladders.Results The standard allelic ladder for DXS8378 locus was obtained,with which the genetic polymorphisms of DXS8378 locus in three Chinese populations in Yunnan were studied.Conclusion The standard ladder made by this method is excellent,and DXS8378 is powerful for forensic practice in Chinese population.

Objective To investigate the expression levels of miRNA in kidney biopsies of child patients with nephrotic syndrome and the possibility of miRNA as potential markers in differentiating the pathologic subtypes of nephrosis.Methods Kidney biopsy specimens from 41 child patients with nephrotic syndrome,including 22 with nesangial proliferative glomeplonephritis (MsPGN),8 with minimal change disease (MCD) and 11 with endocapillary proliferative glomerulonephritis (ECPGN),were collected,and adult nephridial tissues from 8 patients without renal inadequacy were selected as controls.The expression levels of miR-191,miR-151-3p,miR-150,miR-30a-5p and miR-19b in nephridial tissues were detected by RT-qPCR,and their correlations with renal function related parameters were analyzed.Results Compared with the controls,the miR-191 levels in kidney tissues of child patients with nephrotic syndrome increased significantly (P ＜ 0.01),while the miR-151-3p levels decreased obviously (P ＜ 0.01).The expression levels of miR-150 in MCD patients were significantly lower than those in MsPGN and ECPGN patients and the controls (P ＜ 0.05).The expression levels of these miRNAs were positively correlated with serum IgG,TP and Cr levels,but negatively with serum TC levels (P ＜0.05).Conclusion The expression levels of miRNA in kidney tissues of child patients with nephrotic syndrome are related to pathological typing of nephrosis,and miR-150 may be a potential marker which may differentiate MCD from other subtypes of nephrosis.

Objective:To investigate the efficacy and safety of Rituximab (RTX) in treating children with refractory steroid-resistant nephrotic syndrome (SRNS).Methods:The clinical data of 10 children with refractory SRNS receiving RTX in the Department of Pediatrics, Jinling Hospital from September 2013 to March 2018 were analyzed retrospectively.Results:The age of onset of 10 children (including 5 males and 5 females) was (4.47±2.75) years old.The renal biopsy showed focal segmental glomerular sclerosis in 5 cases (50%), minimal change nephropathy in 3 cases (30%), IgM nephropathy in 1 case (10%), and mesangial proliferative glomerulonephritis in 1 case (10%). Ten children received RTX treatment (1 or 4 doses; 375 mg/m 2 once; maximum: 500 mg) at the age of (6.74±2.62) years old.There were 8 patients (80%) receiving a single dose of RTX, 1 patient (10%) receiving 3 doses, and 1 patient (10%) receiving 8 doses.The follow-up time was 11.93 (5.17, 25.66) months.The remission rates at the 3-month follow-up, 6-month follow-up and last follow-up were 30% (3 patients), 40% (4 patients), and 40% (4 patients), respectively.The 24-hour urinary proteinuria and serum albumin levels were improved in 10 children after RTX treatment, but there were no significant statistical difference(all P>0.05). No significant difference was found in humoral immunity and renal function before and after RTX treatment (all P>0.05). During the treatment and follow-up, 3 patients (30%) developed infusion reaction, 2 patients (20%) showed severe pulmonary infection, and 1 patient (10%) died of severe pulmonary infection. Conclusions:RTX is effective in treating some children with refractory SRNS, and a long-term follow-up should be conducted to prevent infection.

Objective Respiratory viruses are the most common pathogens to cause respiratory tract infection in infants and children.The aim of the study was to establish a luminex-based molecular assay for rapid detection of four kinds of common respiratory viruses and provide measures for effective prevention and control . Methods 120 throat swab samples from patients with acute respiratory tract infection were collected in our hospital as disease group.30 normal specimens were used as control group .Specific up-stream and downstream primers , hybridization probes and super prim-ers were designed on the basis of conserved sequences of Influenza A and B viruses( FluA, FluB), respiratory syncytial virus types A and B ( RSVA, RSVB ) from available respiratory-virus sequence data-base.Recombinant plasmid and in vitro transcription RNA positive reference substances were established respectively .The testing sys-tem of Tem-PCR combined with luminex xMAP was built by amplification and optimization of hybridization .Comparative analysis were made between the detection results of the above method and those of single viral gene real -time PCR assay and luminex xTAG assay re-spectively. Results Rapid molecular assay was established to specifically detect the four kinds of respiratory viruses (FluA, FluB, RSVA and RSVB) with the sensitivity of 10 copies/μL.Rapid molecular assay and single viral real-time PCR assay were utilized to de-tect the throat swabs ( n=120 ) from suspected patients , the positive result of the former was 31 .7% ( 38/120 ) and the latter was 29.2%(35/120).The consistency test result indicated the two methods were consistent without a significant difference (k>0.7). Several samples were detected by luminex xTAG assay simultaneously , in which good consistency and significant difference were found in two assays by statistical analysis (k>0.6). Conclusion Preliminary clinical application has confirmed the novel molecular assay is sensitive, specific and rapid in simultaneous detection of FluA , FluB, RSVA and RSVB respiratory viruses , which provides experi-mental basis for accurate diagnosis of infected pathogens at early clinical stage .

Objective To investigate the effect of ambient temperature on body mass, thermogenic activity and un-coupling protein-1 ( UCP1) content of brown adipose tissue ( BAT) in tree shrews ( Tupaia belangeri) , and to provide the-oretical basis for establishing tree shrews model of obesity.Methods Forty healthy adult tree shrews with similar body mass were uesd in our experiment.The tree shrews were divided into five groups (n=8):control group (0 d), the ani-mals were maintained under 25 ±1℃ and 12L:12D ( light : dark, lights on 08:00) photoperiod; and the animals were maintained under 5 ±1℃and 12L:12D photoperiod for 7 d, 14 d, 21 d and 28 d groups, respectively.At the end of ex-periment, the changes of body mass, nonshivering thermogenesis (NST), BAT mass and uncoupling protein 1 (UCP1) con-tent were determined.Results Compared with the control group (0 d), the body mass, NST, BAT mass and UCP1 con-tent of the cold acclimation groups were improved significantly, the BAT color also obviously deepened, and after cold accli-mation for 28 d, the body mass, NST, BAT mass and UCP1 content were increased by 26.32%, 20.65, 53.85%and 43%, respectively.Apparently, the UCP1 content was significantly positively correlated with BAT mass and NST.Conclusions BAT proliferation may be induced and UCP1 expression upregulated by cold acclimation in Tupaia belangeri, therefore, en-hancing the thermogenic activity of brown adipose tissue to increase energy expenditure.We would speculate that BAT might be used as a target organ for treatment of obesity by energetic approach in the future.

Objective To evaluate the value of low-dose spectral CT imaging combined with ASIR recon-struction in differenting peripheral lung cancer from inflammatory mass. Methods We retrospectively analyzed the imaging data of 147 cases with 165 pulmonary nodules or masses underwent enhanced CT scans with spectral imag-ing mode from June 2015 to May 2016. Spectral curves,normalized slope rates,iodine-water concentration were measured on the lesions in arteral phase images. The differences of the spectral characteristic parameters were eval-uated statistically by indepengdent samples t test. Results Among 165 pulmonary nodules or masses,including 68 inflammatory mass and 97 peripheral lung cancer. In the arterial phase,the iodine concentration,water concentra-tion and normalized spectral curves rates of peripheral lung cancer were 10.93 ± 5.12,1033.96 ± 9.74,0.998 ± 0.66,which was obviously higher than those in inflammatory mass in 5.29 ± 0.96,1028.85 ± 9.31,0.620 ± 0.16. There were significant difference between peripheral lung cancer and inflammatory mass(0.000 ,0.008 ,0.001 in P values). Conclusion The iodine concentration,water concentration and normalized spectral curves rates has high value for differential diagnosis of peripheral pung cancer and pneumonia mass in the arterial phase using low-dose spectral CT imaging combined with ASIR reconstruction.

Objective:To analyse the clinical and prognosis of C1q deposition in children with primary membranous nephropathy (PMN).Methods:A retrospective analysis was conducted in 177 children with PMN who were diagnosed by renal biopsy in the Eastern Theater Cornmand General Hospital from July 2005 to September 2013.Patients were divided into C1q deposit group and C1q non-deposit group according to the immunofluorescence staining of C1q.Clinical and pathological characteristics, treatment response, and long-term renal prognosis were compared between the 2 groups.Results:A total of 177 pediatric patients with PMN were included, involving 98 boys and 79 girls with a median age of 192.0 months.During an follow-up of (52.4±35.6) months, 7 cases(4.0%) progressed end-stage renal disease (ESRD), and 14 cases(7.9%) developed ESRD or renal dysfunction.The blood IgG level of C1q deposit group was higher than that of C1q non-deposit group [(5.10±2.51) g/L vs.(4.34±2.10) g/L, t=2.110, P=0.036]. The frequency of glomerular C4 deposits in C1q deposit group was significantly higher than that of C1q non-deposit group (34.7% vs.2.9%, χ2=32.567, P<0.001). The Kaplan-Meier survival analysis showed that there were no differences in cumulative renal survival rate of ESRD ( P=0.561) and cumulative incidence rate of remission ( P=0.291) between groups.The Logistic regression analysis demonstrated that C1q deposition was not correlated with treatment responses ( P=0.587). Univariate COX regression analysis demonstrated that the male gender ( HR=8.578, 95% CI: 1.120-65.689, P=0.039) and no remission ( HR=0.053, 95% CI: 0.017-0.171, P<0.001) were risk factors for renal dysfunction in children with PMN.Multivariate COX regression analysis reveled that no remission ( HR=21.858, 95% CI: 5.595-85.387, P<0.001) and C1q deposition ( HR=0.116, 95% CI: 0.023-0.584, P=0.009) were independent risk factors for renal dysfunction in children with PMN. Conclusions:C1q deposition was an independent risk factor for renal dysfunction in children with PMN.The classical pathway does occur in some PMN patients, which plays an essential role in mediating kidney injury.

Objective: To explore the expression of the A_2A adenosine receptor and the inflammatory cytokines interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) in human degenerative nucleus pulposus (NP) cells after they have been treated with a pulsed electromagnetic field (PEMF). Methods: Human degenerative NP cells were cultured in vitro and treated using an 0.8mT PEMF with a pulse frequency of 50Hz. The pulse width was 150μs and the exposure time was 30min, repeated 5 times at 12 hour intervals. The expression of the A_2A adenosine receptor in NP cells was determined using western blotting and reverse transcription polymerase chain reactions. The expression of the inflammatory cytokines IL-1β and TNF-α were detected using enzyme-linked immunosorbent assays (ELISA). The human degenerative NP cells were also treated with an antagonist and agonist of the A_2A adenosine receptor, and the expression of IL-1β and TNF-α were also determined using ELISA. Results: After the PEMF treatment the expression of the A_2A adenosine receptor increased significantly, while the expression of IL-1β and TNF-α decreased significantly. However, the A_2A adenosine receptor antagonist reversed the inhibitory effect of the PEMF on the expression of IL-1β and TNF-α, while the agonist played an opposite role. Conclusion A PEMF can significantly inhibit the expression of IL-1β and TNF-α in human degenerative NP cells, which could be related to up-regulation of the expression of the A_2A adenosine receptor in those cells.

Objective:To analyze the clinical and prognosis of primary membranous nephropathy (PMN) in children with positive glomerular M-type phospholipase A2 receptor (PLA2R).Methods:A total of 69 children diagnosed with PMN by renal biopsy admitted to the Department of Pediatrics of Eastern Theater Command General Hospital from January 2006 to December 2018 were retrospectively analyzed, including 40 males and 29 females, with an average age of 14.86 years.According to the immunofluorescence of renal pathology, they were divided into PLA2R positive group and PLA2R negative group.Pathological features between 2 groups were compared by the t test, Mann- Whitney U test and Chi- square test.Kaplan-Meier method was used to compare the long-term renal survival rate and cumulative remission rate between 2 groups. Results:A total of 69 pediatric PMN patients were included.The po-sitive rates of serum anti-PLA2R antibody and positive expression of PLA2R in renal tissues were 53.6% (37 cases) and 82.6% (57 cases), respectively.The proportion of children with clinical manifestations of large proteinuria [55 cases(96.5% ) vs.9 cases(75.0%), P=0.034] was significantly higher in the PLA2R positive group than that of the PLA2R negative group.Blood urea nitrogen level was significantly higher in the PLA2R positive group than that of PLA2R negative group[1.14(0.93, 1.54) mg/L vs.0.80 (0.44, 1.18) mg/L, P=0.049], while estimate glomerular filtration rate(eGFR) [162.26 (139.81, 185.53) mL/(min·1.73 m 2) vs.199.52 (157.58, 212.01) mL/(min·1.73 m 2), P=0.034] and serum IgG [3.58 (2.50, 5.43) g/L vs.5.14 (4.35, 6.03) g/L, P=0.016] were significantly lower.The cumulative remission rate was significantly higher in the PLA2R negative group than that of PLA2R positive group ( P<0.001). The 24 h urinary protein ≥50 mg/kg ( HR=0.119, 95% CI: 0.021-0.595, P=0.010)was an independent risk factor for renal prognosis, and PLA2R( HR=0.263, 95% CI: 0.125-0.551, P<0.001) and 24 h urinary protein ≥50 mg/kg ( HR=0.568, 95% CI: 0.125-0.551, P=0.041)were independent predictors of urinary protein remission.PLA2R ( HR=1.020, 95% CI: 0.698-1.682, P=0.656)was not associated with renal prognosis. Conclusions:The severity of PMN in children with positive PLA2R was higher than that in those with negative PLA2R.The long-term cumulative remission rate of PLA2R negative children with PMN was higher than that of PLA2R positive children.