High-resolution melting analysis ( HRMA/HRM ), a simple, rapid, flexible, inexpensive closed tube approach with high sensitivity and specificity has been one of the most widely used molecular diagnostic techniques in clinicalas well as in research settings .Recently, rapid development ofinstruments , DNA dyes and analysis software significantly enhance the sensitivity , specificity and accuracy of HRM,providing a fast, efficient and economic molecular diagnostic platform for molecular diagnosis of inherited disease , molecular profiling and target therapy of cancer , identification of pathogen , as well as individualized medicine.

Objective To explore the expression status of carbonic anhydrase Ⅲ (CA Ⅲ) from quadriceps femoris muscle in two kinds of muscle clinical phenotype (skeletal muscle atrophy group and skeletal muscles non-atrophy group) of chronic obstructive pulmonary disease (COPD).Methods Totally 37 inpatients from our hospital,were divided into 11 patients without COPD and 26 patients with COPD,in addition,according to body mass index,fat free mass index and quadriceps cross-section diameter,patients with COPD were divided into 14 skeletal muscles non-atrophy patients (SMNA)and 12 skeletal muscle atrophy patients (SMA).CA Ⅲ concentration of femoris quadriceps specimens was quantitatively determined using Western blot methods,CA Ⅲ mRNA expression levels of femoris quadriceps specimens were also quantitatively measured using RT-PCR,then compared among the 3 groups.Results There was significant difference in CA Ⅲ quantitative concentration and CA Ⅲ mRNA expression level in each group (P ＜ 0.05),further more,CA Ⅲ concentration expression level was significantly higher (P ＜ 0.01)in SMA group (1.260 ± 0.068) than in SMNA group (1.110 ± 0.014),the latter was significantly higher (P ＜ 0.01) than in the control group (1.000 ± O.062).CA Ⅲ mRNA expression level was significantly higher(P ＜ 0.01) in SMNA group(2.170 ±0.412) than in the control group(1.000 ±0.115),and was significantly lower than in SMA group (3.770 ± 0.788 ; P ＜ O.01).CA Ⅲ concentration and CA Ⅲ mRNA expression level increased at equal pace in SMNA group and SMA group,however,CA Ⅲ quantitative concentration and CA Ⅲ mRNA expression level were inconsistent in the two groups.Conclusion The expression status of CA Ⅲ in quadriceps femoris muscle was different in two kinds of muscle clinical phenotype of COPD.

Objective To develop a rapid method based on ARMS, named four primers allele-specific PCR(4p-AS PCR), to detect the most common non-deletional ?-thalassemia mutation Hb Constant Spring(Hb CS) by PCR technique. Methods The 4p-ASPCR was used to detect the Hb CS mutation of ?-thalassemia in 38 DNA samples of Hb H disease patients and using PCR-RE and DNA sequencing to confirm the results. Results Among the 38 Hb H disease patients 15 cases was revealed to carry Hb CS, 16 cases were deletional Hb H, and 7 cases need to be defined.Conclusion A simple, rapid and reliable method, named 4P-ASPCR, to detect Hb CS mutation have been developed. It is also may be useful in screen other point mutations such as Hb Quong Sze.

To select appropriate descriptors for response of the patient-reported outcome (PRO) scale for the main symptoms of patients with chronic obstructive pulmonary disease (COPD) complicated with pulmonary heart disease.

Objective To compare and analyze MRI characteristics of medical patellofemoral ligament(MPFL) injury patterns in adolescents and adults after acute lateral patellar dislocation.Methods MR images of 42 adolescents and 45 adults after acute lateral patellar dislocation were retrospectively reviewed.Routine MR scanning was performed in axial,sagittal and coronal planes,including T1 WI,T2 WI and fat saturation sequence.The x2-tests were performed to analyze the MPFL injury patterns between adolescent group and adult group.Results The prevalence rate of MPFL injury was 97.6％ (41/42) in adolescent group after acute lateral patellar dislocation,including 57.1％ (24/42) of complete tear and 40.5％ (17/42) of partial tear.The prevalence rate of MPFL injury was 100.0％ (45/45) in adult group,including 64.4％ (29/45) of complete tear and 35.6％ (16/45) of partial tear.There were no statistical difference between adolescents and adults in prevalence rate of MPFL injury,complete MPFL tear,partial MPFL tear (x2 =1.084,0.486,0.223 ; P ＞ 0.05).The prevalence rate of MPFL injury at femoral attachment,patellar attachment and midsubstance were 31.0％ (13/42),78.6％ (33/42) and 26.2％ (11/42) respectively in adolescent group,whereas they were 64.4％ (29/45),40.0％ (18/45) and 15.6％ (7/45) respectively in adult group.It occurred at multiple sites in 33.3％ (14/42) of adolescents,and in 15.6％ (7/45) of adults.There were significant differences between adolescents and adults in the prevalence rate of MPFL tear at femoral attachments and patellar attachments (x2 =9.759,13.324 ; P ＜ 0.05),but there was no statistical difference at its midsubstance (x2 =1.497,P ＞ 0.05).Although not statistically significant (x2 =3.749,P ＞ 0.05),MPFL tear at multiple locations were seen more in adolescents than in adults (33.3％ vs.15.6％).Conclusion The injury degree and distribution of MPFL are similar in adolescents and adults after acute lateral patellar dislocation,but there is significant difference in the site of MPFL injury distribution between adolescents and adults.MPFL is most easily injured at femoral attachment,secondly at patellar attachment in adults,while MPFL is most easily injured at patellar attachment,secondly at femoral attachment in adolescents.Adolescents are more likely to have multiple-site MPFL injury than adults.

Objective To compare HER-2 state in breast cancer tissue deteced by fluorescent in situ hybridization (FISH) and immunohistochemistry (IHC) and analyze their correlation. Methods HER-2/neu protein expression and gene amplification were detected by FISH and IHC in 56 newly-diagnosed cases of female breast cancer from July 2008 to July 2009. Results Of the 56 patients, HER-2 protein expression (-), (+), (++), (+++) was 9 cases (16.1%), 29 cases (51.8%), 11 cases (19.6%) and 7cases (12.5%) respectively; 26 cases (46.4%) had HER-2 gene amplification while 30 cases (53.6%) didnt have. Type of HER-2 gene amplification was mainly HER-2(++) and HER-2(+++), and according gene amplification rate was 72 7% and 100%. HER-2 (+) gene amplification rate was 37.9 %(11cases) and no gene amplification was found in HER-2(-) tissue. The HER-2 positive rate using two methods had significant difference(χ2=19.778,P<0.01). HER-2(-) and HER-2(+++) had good consistency with the FISH results(Kappa=0.969),but HER-2(+) and HER-2(+ +) were poorly consistent with the FISH results(Kappa=0.271). Conclusions IHC is the preliminary screening method for detection of HER-2 expression. HER-2(-) and HER-2(+++) have good consistency with the gene amplification, and can guide clinical treatment. Some patients with HER-2(+) and HER-2(++) have HER-2 gene amplification. FISH is needed for targeted therapy.

Objective To determine the clinical implications of internal mammary node biopsy for neoplasm stage,treatment,and prognosis in patients with breast cancer.Methods Internal mammary node biopsy via intercostal space was performed in 229 cases of breast cancer.Anatomical location of internal mammary nodes was recorded.Results Internal mammary node biopsy was successfully finished in 220 patients.There were 56 cases (24.45％ ) with internal mammary nodes metastasis,126 cases (55.02％ ) with axillary nodes metastasis,43 cases (34.13％ ) with regional metastases in both the axillary and internal mammary lymph nodes and 13 cases ( 12.62％ ) with internal mammary node metastasis only.Internal mammary node metastasis rate in patients with the number of positive axillary nodes ≥4 was 49.32％ (36/73).pN stage migration was seen in 56 patients with positive internal mammary nodes.There was no statistic relation between internal mammary nodes metastases and tumor location ( x2 =0.661,P =0.719).70.7％ patients with medial/central tumors and 50.7％ patients with the number of positive axillary nodes ≥4 were free from internal mammary node radiotherapy on account of internal mammary node biopsy.There was no complication such as pneumothorax or haemorrhagia.Conclusions Internal mammary node biopsy from intercostal space is a reliable surgical technique and can improve pN stage in some breast cancer patients.With internal mammary node biopsy,patients with a negative internal mammary node can be prevented from radiation to internal mammary nodal areas.

Objective To explore the best operation method in the patients with papillary thyroid microcarcinoma.Methods A total of 139 patients with papillary thjroid microcarcinoma were treated at our institute between Jan.2000 and Jan.2010.The data on the clinicopathological characteristics of patients and treatment were collected.Results The mean tumor size was (0.45 ± 0.24) cm.Of the 139 patients,30.2％ had multifocal tumors,19.4％ had bilateral tumors,42.4％ neck lymph node metastases.The number of eases of lymph node metastasis in level Ⅵ,Ⅱa,Ⅲ and Ⅳ was 58(41.7％),3(2.2％),10(7.9％),5(3.6％),respectively.Only one had lymph node metastasis in Level Ⅲ Microscopic extrathyroid extension was associated with neck lymph node metastases in papillary thyroid microcareinoma patients (x2 =38.39,P ＜0.001).No one developed permanent hypoparathyroidism.The median follow-up time was 103 (range,30-154) months,and local recurrence in the thyoid was diagnosed in 2 patients who underwent hemi-or subtotal thyroidectomy.Follow-up of 10 years was done in 16 cases,and the survival rate of 139 patients for 10-year was 100％.Conclusions It suggested that patients with papillary thyroid microcarcinoma has uniform clinicopathologic characteristics and the pattern of lymph node metastasis from those with papillary thyroid carcinoma.Total thyroidectomy plus level Ⅵ dissection is the optimal surgical treatment of papillary thyroid microcarcinoma.

Objective To explore the pattern of cervical nodal metastasis and the clinical significance of total thyroidectomy plus functional neck lymphadenectomy in papillary thyroid carcinoma patients. Methods Clinical and pathological data of 172 patients with papillary thyroid cancer who underwent total thyroidectomy plus functional neck lymph node dissection at Jinan Military General Hospital were retrospectively reviewed, including patient demographics, extent of surgery, parathyroid hormone level,recurrence, tumor pathology, such as tumor size, multifocality, capsular invasion, vascular invasion,extrathyroidal extension, and lymph node status. Results Of the 172 functional neck dissection patients (47 ambilateral), the incidence of lymphonodus metastasis in regions Ⅵ, Ⅳ and Ⅲ was 96. 3% ,78. 5%and 62.1% respectively. Rate of nodal metastasis was higher in patients with extracapsular invasion than in patients with no invasion ( P ＜ 0. 05 ). Serum parathyroid hormone levels significantly decreased immediately postoperatively in total thyroidectomy plus functional neck dissection and remained low for several weeks thereafter ( P ＜ 0. 01 ). The 5-, 10- and 15-year survival rate was ( 98. 83 ± 0. 82) %, (98. 23 ± 1.02 ) % and (96. 42 ± 1.43 )%, respectively. Conclusions Therapeutic neck lymph node dissection for papillary thyroid cancer is recommended for cervical nodal metastasis patients. Total thyroidectomy plus functional neck dissection is important in the treatment of papillary thyroid cancer.

Objective To evaluate the feasibility and clinical efficacy of intra-abdominal exposure instruments in laparoendoscopic single-port nephrectomy(LESS-N).Method From February 2012 to July 2016,61 cases of LESS-N were performed in our center.There were 34 males and 27 females with a mean age of (60.3 ± 9.4) years old (ranging 36-72 years old).There were thirty-nine cases of renal tumors and twenty two cases of nonfunctioning kidney.The patients were divided into two groups.Group A included 39 cases that underwent conventional LESS-N (22 radical nephrectomy/17 simple nephrectomy).Group B included 22 cases that underwent intra-abdominal exposure instruments assisted LESS-N (17 radical nephrectomy/5 simple nephrectomy).The perioperative and postoperative data were collected and analyzed retrospectively.Results All the procedures of these two groups were completed successfully.In Group A,four patients were added one 5 cm additional trocar and two patients were converted to open surgery.No additional trocars or conversion to open surgery were needed in Group B.For LESS radical nephrectomy,there were no significant differences of mean tumor diameter (5.7cm vs.5.4 cm,P =0.65) between two groups.The average operative time was (95.1 ± 43.9) min in Group B which was lower than that in Group A (127.4 ± 61.9) min (P ＜ 0.01).The mean renal vascular processing time was declined from (25.4 ± 10.1)rmin in Group A to (18.8 ± 8.9)min in Group B (P ＜ 0.05).The mean estimated blood loss was (128.6 ± 51.1) ml in Group A and (98.7 ±-57.6) ml in Group B (P ＜ 0.05).No severe intraoperative and postoperative complications occurred in both group.Conclusions Intra-abdominal exposure instruments are feasible and effective for LESS-N.This system may shorten the operation time,reduce the amount of bleeding and improve surgical accuracy.