Objective To resolve the problem of the accuracy and standardization of STR-PCR typing in forensic practice,construct DXS6804 allelic ladder by molecular clonning and apply them in a population study on the Pumi population in Yunnan,China.Methods PCR was used to produce several different allelic fragments of the locus.After cloning the PCR products,the recombinant plasmids were sequenced.Then we denominated them and used them as template for re-amplification to generate the locus standard ladder.Results The sequencing results confirmed that the size and the construction of the inserts were correct.The genetic polymorphisms of this locus in Yunnan Pumi population of China were studied.Two off-ladder alleles of DXS6804 locus were found.Conclusion This method is of high value for forensic DNA typing to construct standard ladders.DXS6804 is robust for genetic research and forensic application.

Objective To examine the association between the polymorphisms of brain-derived neurotrophic factor (BDNF)gene with heroin dependence.Methods Genomic DNA was isolated from the venous blood leukocytes of 308 unrelated patients with heroin dependence and 31 7 healthy individuals.Seven single nucleotide polymorphisms (SNPs)were genotyped using MassARRAY system.Data were analyzed using HaploView 4.0 and SPSS 20.0 software.Results There was a significant difference in the genotype frequency of rs6265 between heroin dependence group and healthy control group (χ2 =1 5.1 5 1,P =0.001).The rs6265 G allele was significantly higher than in controls (χ2 =9.864,P =0.002,OR =1.429,95% CI =1.143 -1.786).Furthermore,there was also a significant difference in the genotype frequency of rs13306221 between heroin dependence group and control group (χ2 =7.699,P =0.006).The rs13306221 G allele was significantly higher than in controls (χ2 =7.137,P =0.008,OR =0.539,95% CI =0.340-0.853).Strong linkage disequilibrium (LD)was observed in one block (D’> 0.9;r 2 >0.8),and significantly less G-G haplotype frequency of block 1 (χ2 =4.546;P =0.033)was found in heroin dependence group. Conclusion Our findings support the role of BDNF rs6265 and rs13306221 polymorphisms in heroin dependence and may guide future studies to identify other genetic risk factors for heroin dependence.

Objective To study the effects of exogenous phosphocreatine (CP) on brain injury after cardiopulmonary resuscitation (CPR) in rats.Methods A total of 160 male adult SD rats were randomly ( random number) divided into 4 groups:sham-operation control group ( group A),CPR group ( group B),low-dose CP group ( group C),high-dose CP group ( group D),and each group was further divided into 5 subgroups (n =8) as per study at different intervals,0.5,3,6,12 and 24 h after restoration of spontaneous circulation (ROSC) in groups B,C and D or after tracheotomy in group A.Cardiac arrest (CA) was induced by using asphyxia to establish CPR model in group B,C and D.The CP0.5 g/kg used in group C or CP 1.0 g/kg used in group D was injected into femoral vein at beginning of ROSC.Rats in each subgroup were sacrificed and the tissues of frontal lobe of brain of rats were taken at different intervals.The levels of adenosine triphosphate (ATP),adenosine diphosphate (ADP) and adenosine monophosphate (AMP) in cerebral cortex were measured by high performance liquid chromatography (HPLC),and values of total adenine nucleotides ( TAN ) and energy charge ( EC ) were detected.The activities of Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase in cerebral cortex were assayed by spectrophotometric method. The pathological changes of cerebral cortex were observed under optical microscope.The experimental data were processed with analysis of variance by using SPSS 16.0 package. Results Compared with group A,the levels of ATP,TAN,EC,Na + -K + -ATPase,Ca2+ -Mg2+ -ATPase were lower ( P ＜ 0.05 or P ＜ 0.01 ) at each interval in groups B and C,and at intervals of 0.5,3,6,12 h in group D,and the levels of AMP were higher (P ＜ 0.01 ) at each interval in group B and at intervals of 0.5 h and 3 h in groups C and D.Compared with group B,the levels of ATP,TAN,EC,Na + -K + -ATPase,Ca2 + -Mg2+ -ATPase were higher ( P ＜ 0.05 or P ＜ 0.01 ),and the levels of AMP were lower ( P ＜ 0.05 or P ＜ 0.01 ) at intervals of 6,12 and 24 h in groups C and D.Compared with group C,the levels of ATP at interval of 24 h and TAN,Na +- K + -ATPase,Ca2 + -Mg2 + -ATPase at intervals of 6,12 and 24 h were higher in group D ( P ＜ 0.05 or P ＜ 0.01 ).There were severe pathological changes in cerebral cortex in group B,and mild changes in groups C and D. Conclusions There was obvious energy metabolism disorder after CPR in rats.Treatment with exogenous CP could increase the levels of ATP and activities of ATPase,alleviate pathological changes,especially in high-dose,and mitigate injury in cerebral cortex after CPR in rats.

Objective To observe the expression changes in apoptosis-related microRNA(miRNA) in cerebral cortex after cardiac arrest-cardiopulmonary resuscitation(CA-CPR)in rats and explore the factors that may affect the mechanism of CPR. Methods 24 clean male Sprague-Dawley(SD)rats were randomly divided into three groups,the normal control group,sham operation group and CA-CPR group(each n=8). The animal model of CA induced by asphyxia was established and CPR was performed. In the normal control group,no special management was performed. In the sham operation group,only abdominal cavity anesthesia,tracheotomy,vascular puncture and electrocardiogram(ECG)were performed without clamping the trachea and resuscitating. Normal feeding in normal control group and 24 hours after tracheotomy in sham operation group,at 24 hours after recovery of spontaneous circulation(ROSC)in CA-CPR group,cerebral cortex specimens were obtained for detection of the expression of miRNA by using real time fluorescence quantitative reverse transcription - polymerase chain reaction(RT-PCR). Flow cytometry(FCM)was used to detect the neurocyte apoptotic rate. Results Compared between normal control and sham operation groups,there were no significant differences in the expression of apoptosis-related miRNA and neurocyte apoptosis rate of cerebral cortex(both P>0.05). Compared with sham operation group,in CA-CPR group, 16 miRNA expressions were up-regulated,including Let-7c,miR-15a,miR-21,miR-24,miR-29,miR-29b, miR-34a, miR-103, miR-200a, miR-200b, miR-200c, miR-210, miR-326, miR-338-3p, miR-494 and miR-497,and there were 22 down-regulated,being Let-7a,Let-7b,Let-7d,Let-7e,miR-19a,miR-19b-1, miR-20a,miR-20b,miR-23a,miR-23b,miR-25,miR-98,miR-107,miR-122a,miR-125a,miR-125b, miR-145,miR-181a,miR-181c,miR-335,miR-384-5p and miR-422a. Eight miRNA had significant changes at 24 hours after ROSC,in which miR-15a,miR-21,miR-34a,miR-497 were up-regulated respectively for 6.831±2.625,8.122±3.442,5.349±2.010,6.590±3.689 times,and miR-125b,miR-145,Let-7a,Let-7e were down-regulated respectively for 0.122±0.039,0.199±0.096,0.191±0.069,0.160±0.082 times. The apoptosis rate of cerebral cortex was increased significantly in CA-CPR group〔(32.23±5.31)%〕compared with that in normal control group〔(3.66±1.34)%〕and sham operation group〔(4.98±1.84)%,both P<0.01〕. Conclusions In early period after CA-CPR,obvious neurocyte apoptosis may be found in brain tissue of rats,and in the mean time, changes in apoptosis-related miRNA expression in cerebral cortex occur. The various types of miRNA with significant changes possibly play important roles in cerebral protection after CA-CPR in rats.
down-regulated respectively for 0.122±0.039,0.199±0.096,0.191±0.069,0.160±0.082 times. The apoptosis rate of cerebral cortex was increased significantly in CA-CPR group〔(32.23±5.31)%〕compared with that in normal control group〔(3.66±1.34)%〕and sham operation group〔(4.98±1.84)%,both P<0.01〕. Conclusions In early period after CA-CPR,obvious neurocyte apoptosis may be found in brain tissue of rats,and in the mean time, changes in apoptosis-related miRNA expression in cerebral cortex occur. The various types of miRNA with significant changes possibly play important roles in cerebral protection after CA-CPR in rats.

Objective To investigate the nonsentinel lymph nodal (NSLN) status in 1-2 sentinel lymph node positive early breast cancer and to compare with that in Z0011 ALND group.Methods A total of 234 T1,T2 invasive breast cancer with 1-2 positive sentinel lymph nodes were collected in Henan Tumorr Hospital from Jul 2010 to Aug 2014.The data was compared with that of Z0011 ALND group.Results 234 female patients were enrolled.76 cases had non sentinel lymph node metastases.The proportion of 1 and 2 sentinel lymph node metastases was 75.2％ and 24.8％,respectively.The number of positive sentinel lymph nodes and that of negative ones were independent predictors of the non sentinel lymph node status (all P ＜ 0.05).Menstrual status,hormone receptor,Ki67 level and HER2 were not associated with nonsentinel lymph node metastasis (all P ＞ 0.05).They had nonsentinel lymph node positive rate (P ＞ 0.05) similar to that of Z0011 ALND group.Conclusious The number of sentinel lymph nodes and negative sentinel lymph nodes were associated with nonsentinel lymph node metastasis in the 1-2 sentinel lymph positive early stage breast cancer.Those patients had similar positive nonsentinel lymph node rate to that of Z0011 ALND group.

Objective:To investigate the correlation between ipsilateral metastatic supraclavicular lymph nodes(ISLN) pathologically complete response (spCR) and the prognosis in breast cancer after neoadjuvant chemotherapy(NAC) and lymphadenectomy.Methods:Clinical data of 234 breast cancer patients with ISLN at the time of diagnosis and receiving supraclavicular lymph node dissection following NAC were retrospectively analyzed.Results:All patients were female, with a median age of 52 years. The spCR rate was 52.6%. Multivariate analysis showed that the expression level of Ki67, the pathological status of breast and axillary nodes and the dissection number of supraclavicular lymph node were independent relate factors for spCR (all P<0.05). After a median follow-up of 16.6 months, the risk of recurrence and metastasis in the spCR group was 51%. It was lower than that in the non-spCR group ( HR=0.49, 95% CI 0.27-0.89, P=0.020). It was mainly manifested in hormone receptor negative patients. Supraclavicular spCR was an independent prognostic factor for DFS. Conclusions:The expression level of Ki67, pathological state of breast and axillary node and the dissection number of supraclavicular lymph node were independent related factors of spCR, which was also an independent predictor of DFS.

OBJECTIVETo investigate the predictive factors affecting sentinel lymph node status in early breast cancer patients.

METHODSClinicopathological data of 1 038 patients with early breast cancer, who underwent sentinel lymph node biopsy in Henan Tumor Hospital between July 2010 and August 2013, were reviewed. Logistic regression analysis was performed to identify the relevance of clinicopathological features with sentinel lymph node metastases.

RESULTSThis group was consisted of 1 038 female patients with an average of 48.6 years. Positive sentinel lymph nodes were found in 22.9% (238/1 038) of the patients. The average number of sentinel lymph nodes removed by surgery was 3.8. Tumor size, tumor location, histopathology, ER/PR status and Ki-67 level were significantly correlated with SLN metastasis(P < 0.05 for all). All the above factors but Ki-67 level were significant independent predictors for SLN metastasis(P < 0.01 for all).

CONCLUSIONNegative hormone receptor status, invasive cancer of non-specific histopathological type, tumor size >2 cm, and tumor location in the outer upper quadrat are independent predictive factors of sentinel lymph node metastasis in patients with early breast cancer.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; surgery ; Female ; Humans ; Ki-67 Antigen ; metabolism ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Retrospective Studies ; Sentinel Lymph Node Biopsy ; Tumor Burden ; Young Adult

Objective: To investigate the influence of lumpectomy on axillary lymph node status of breast cancer patients. Methods: The clinical data of 738 invasive breast cancer patients with non-palpable axillary lymph node and sentinel lymph node (SLN) biopsy from November 2011 to August 2013 in Henan Provincial Cancer Hospital were collected and retrospectively analyzed. Among them, 136 patients underwent preoperative lumpectomy (lumpectomy group) and 602 patients underwent puncture biopsy only (biopsy group). The difference of axillary lymph node status and positive ratio of SLN detected by color Doppler ultrasound were compared between these two groups. Results: Among the 738 breast cancer patients, the axillary lymph nodes of 444 (60.2%) cases could be detected by ultrasound. Among them, 92 cases belonged to lumpectomy group, significantly less than 352 cases of biopsy group (P=0.048). Among the patients with ultrasound-visible lymph nodes, the proportion of the biggest diameter of axillary lymph node >1 cm of lumpectomy group or biopsy group was 58.7% (54/92) or 52.8% (186/352), respectively, without significant difference (P=0.316). The proportion of patients with the ratio of long diameter to short diameter <2 of lumpectomy group or biopsy group was 37.0% (34/92) or 38.6% (136/352), respectively, with marginal difference (P=0.768). The positive rate of SLN of lumpectomy group or biopsy group was 23.5% (32/136) or 26.9% (162/602), respectively, without significant difference (P=0.419). The incidence rate of the ultrasound visible axillary lymph nodes of patients whose postoperative time ≤ 7 days or > 7days was 71.1% (64/90) or 60.9% (8/46), respectively, without significant difference (P=0.227). However, the positive rate of SLN of these two groups was 28.9% (26/90) and 13.0% (6/46), respectively, with significant difference (P=0.039). The number of ultrasound visible axillary lymph nodes, the biggest diameter of axillary lymph nodes and the ratio of the long diameter to short diameter <2 were substantially correlated with the positive rate of SLN (P<0.05). Conclusions The incidence rate of ultrasound visible axillary lymph node in the patients with lumpectomy is higher than that of patients with puncture biopsy only. The positive rate of SLN of the patients with a long postoperative time is lower than that of patients with a short postoperative time, even though the axillary lymph nodes are ultrasound visible.