In order to establish a simple and useful way for preimplantation genetic diagnosis (PGD) of chromosomal diseases in general IVF laboratory, the methods that are most commonly used in the embryo biopsy, fixation of blastomere and fluorescence in situ hybridization were compared. The three aspects of PGD were analyzed respectively. There was no significant difference in further development capacity of embryos between mechanical (79.7%) and chemical biopsy group (78.6%) (P>0.05). In this study, more cells were successfully fixed with the Tween/HCL method (93.8%) than with the methanol/acetic acid method (80.5%, P<0.05). There was no significant difference in cytoplasm remains between methanol/acetic acid method and Tween/HCL method (P>0.05). The hybridization efficiency of fluorescence in situ hybridization was 89.5% in successive denaturation method and 90.9% in codenaturation method with the difference being not significant (P>0.05). In conclusion, the mechanical or chemical method, Tween/HCL fixation method and codenaturation fluorescence in situ hybridization method can constitute a simple and useful way for PGD of chromosomal diseases.

Objective: To analyze the influence of semen processing methods on the proportion of the aneuploid sperm by detecting the sperm's chromosome X, Y, 18 using fluorescence in situ hybridization. Methods: Ten patients with mild ohgoasthenosperia, who were received ICSI treatment, were included in this study. Five semen samples of the patients were randomly selected to detect using Swim-up method (A group) and 5 using sperm-grad double-density centrifugation method (B group).Another 5 patients with mild oligo-asthenosperia were as control (C group). The CEPX / Y and CEP18 probe was used to detect the sperm of these 15 patients by fluorescence in situ hybridization. The proportion of aneuploid sperm was compared in three groups. Results: The sex chromosome aneuploid rates were (4.21±2.49)%, (3.24～1.49) % and (2.62±0.89) % in control, A and B groups. The rates of aneuploid chromosome 18 were (3.00±1.22)%, (2.00～1.22)% and (2.00±1.22)% in control, A and B groups. There were no significant differences in three groups (P＞0.05). Conclusion: The results showed that the methods of Swim-up and Sperm-Grad double-density gradient centrifugation could select sperms in motility potential and teratospermia,but not in normal chromosome sperms.

Objective To analyze clinical manifestation and investigate therapy effect of re-operation of tricuspid regurgitation (TR) after mitral valve replacement with rheumatic heart disease.Methods Seventeen cases with rheumatic heart disease recurred TR after mitral valve replacement surgery,underwent tricuspid valve surgery again in the Third People's Hospital of Nanchang from January 2000 to December 2011.Of 17 cases,10cases underwent tricuspid valve annuloplasty including 1 case for pure De Vega plasty,9 cases for the valve leaflets forming + artificial valve ring forming.Another 7 cases underwent tricuspid valve replacement surgery including 4 cases for biological valve replacement and 3 cases for mechanical valve.Retrospective analyzed the clinical manifestations,treatment process and condition of prognosis.Results One case was with early postoperative deaths (5.88％,1717),and died of postoperative left ventricular failure.Three cases were postoperative low cardiac output syndrome,2 cases were renal insufficiency,and 2 cases were respiratory insufficiency,all those cases were successfully cured.Sixteen cases were followed up from 3 months to 9 years and 2 cases were lost.Of 14,2 cases were NYHA class Ⅰ,8 cases for grade Ⅱ,4 cases for grade Ⅲ.Conclusion After mitral valve replacement in patients with rheumatic heart disease,TR in patients with reoperation is a suitable choice.Reasonable surgical indications,timing of surgery and good perioperative management are the keys to improve the success rate of surgery.

BACKGROUNDIntrathoracic lymph node metastasis including hilus and mediastinum is one of important prognosis factors of lung cancer. The aim of this study is to investigate the characteristics and patterns of lymph node metastasis in non-small cell lung cancer and to provide evidence for determining the range of lymph node dissection.

METHODSOne hundred and fifty-five patients with non-small cell lung cancer received complete resection combined with systematic lymph node dissection according to the mapping system developed by Naruke.

RESULTSA total of 1553 lymph nodes were dissected from 155 lung cancer patients. The positive ratios of N1 and N2 were 20.0% (31/155) and 38.7% (60/155) respectively. Fifteen patients (9.7%) were found with skipping N2 which were located in lymph node groups 2, 4, 5, 6 and 7 respectively. There was close correlation between lymph node metastasis and T stage (P < 0.01). The metastatic rate of lymph node in different histology had no significant difference. Skipping mediastinal lymphatic metastasis was found more frequently in lung cancer.

CONCLUSIONSLymph node metastasis of non-small cell lung cancer may occur in multiple groups and regions, even in a skipping pattern. Systematic lymph node dissection may be routinely performed in pulmonary resection for non-small cell lung cancer except stage T1.

OBJECTIVETo establish a novel HLA genotyping method for preimplantation genetic diagnonis (PGD) using multiple displacement amplification-polymerase chain reaction-sequencing based technique (MDA-PCR-SBT).

METHODSPeripheral blood samples and 76 1PN, 2PN, 3PN discarded embryos from 9 couples were collected. The alleles of HLA-A, B, DR loci were detected from the MDA product with the PCR-SBT method. The HLA genotypes of the parental peripheral blood samples were analyzed with the same protocol. The genotypes of specific HLA region were evaluated for distinguishing the segregation of haplotypes among the family members, and primary HLA matching was performed between the embryos.

RESULTSThe 76 embryos were subjected to MDA and 74 (97.4%) were successfully amplified. For the 34 embryos from the single blastomere group, the amplification rate was 94.1%, and for the 40 embryos in the two blastomeres group, the rate was 100%. The dropout rates for DQ allele and DR allele were 1.3% and 0, respectively. The positive rate for MDA in the single blastomere group was 100%, with the dropout rates for DQ allele and DR allele being 1.5% and 0, respectively. The positive rate of MDA for the two blastomere group was 100%, with the dropout rates for both DQ and DR alleles being 0. The recombination rate of fetal HLA was 20.2% (30/148). Due to the improper classification and abnormal fertilized embryos, the proportion of matched embryos HLA was 20.3% (15/74),which was lower than the theoretical value of 25%.

CONCLUSIONPGD with HLA matching can facilitate creation of a HLA-identical donor (saviour child) for umbilical cord blood or bone marrow stem cells for its affected sibling with a genetic disease. Therefore, preimplantation HLA matching may provide a tool for couples desiring to conceive a potential donor progeny for transplantation for its sibling with a life-threatening disorder.

Blastocyst ; cytology ; metabolism ; Female ; Genotype ; Genotyping Techniques ; methods ; HLA Antigens ; genetics ; HLA-DQ beta-Chains ; genetics ; HLA-DRB1 Chains ; genetics ; Humans ; Polymerase Chain Reaction ; methods ; Pregnancy ; Preimplantation Diagnosis ; methods ; Reproducibility of Results ; Sequence Analysis, DNA ; methods

Objective:To investigate the effect of body mass index (BMI) on clinical pregnancy and neonatal outcomes in patients with polycystic ovary syndrome (PCOS) during frozen-thawed embryo transfer.Methods:A total of 650 patients with PCOS who received routine in vitro fertilization or intracytoplasmic sperm injection treatment for frozen-thawed embryo transfer from June 2014 to June 2019 in Tianjin Central Hospital of Gynecology Obstetrics were retrospectively analyzed. According to BMI, PCOS patients were divided into group A (18.5≤BMI<23 kg/m 2, n=253), group B (23≤BMI<25 kg/m 2, n=167), and group C (BMI≥25 kg/m 2, n=230). The general information, clinical pregnancy outcomes, pregnancy complications, the incidence of macrosomia and low-birth-weight infants were compared in the three groups, and the influencing factors of neonatal birth weight were analyzed. Results:The embryo implantation rate, clinical pregnancy rate, and ongoing pregnancy rate all showed downward trend with the increase of BMI, but the differences were not statistically significant (all P>0.05). The live birth rate in group C [47.0% (108/230)] was significantly lower than those in groups A and B, with statistical significance ( χ2 =7.43, P=0.024). The late miscarriage rate was higher in group C [9.4% (13/139)] than in groups A and B ( χ2 =7.66, P=0.022). The birth rates of macrosomia in groups B [22.2% (16/72)] and group C [21.1% (16/76)] were significantly higher than that in group A, and the difference was statistically significant ( χ2 =14.15, P=0.001). There was no statistically significant difference in the incidence of gestational diabetes between the three groups ( χ2 =3.81, P=0.149). The incidence of hypertension disorders complicating pregnancy increased with the increase of BMI, and the difference was not statistically significant ( P>0.05). Regression analysis showed that macrosomia was significantly associated with maternal pre-pregnancy BMI and gestational weeks, and the risk of macrosomia increased by 15% (95% CI: 3%-28%) for every increase in maternal BMI. Conclusions:The embryo implantation rate, clinical pregnancy rate, and ongoing pregnancy rate of PCOS patients in frozen-thawed embryo transfer cycles show downward trend with the increase of BMI. Obese patients with PCOS have a significant increase in late miscarriage rate and a significant decrease in live birth rate. The incidence of hypertension disorders complicating pregnancy in PCOS patients in the obese group has an increasing trend, and the birth rate of macrosomia has increased significantly. Therefore, it is recommended that obese women with PCOS lose weight scientifically before pregnancy to improve pregnancy and neonatal outcomes.

In order to establish a simple and useful way for preimplantation genetic diagnosis (PGD)of chromosomal diseases in general IVF laboratory, the methods that are most commonly used in the embryo biopsy, fixation of blastomere and fluorescence in situ hybridization were compared. The three aspects of PGD were analyzed respectively. There was no significant difference in further development capacity of embryos between mechanical (79.7%) and chemical biopsy group (78.6%)(P＞0.05). In this study, more cells were successfully fixed with the Tween/HCL method (93.8%) than with the methanol/acetic acid method (80.5%, P＜0.05). There was no significant difference in cytoplasm remains between methanol/acetic acid method and Tween/HCL method (P＞0.05). The hybridization efficiency of fluorescence in situ hybridization was 89.5% in successive denaturation method and 90.9% in codenaturation method with the difference being not significant (P＞0.05). In conclusion, the mechanical or chemical method, Tween/HCL fixation method and codenaturation fluorescence in situ hybridization method can constitute a simple and useful way for PGD of chromosomal diseases.

Objective:To explore any effect of brain aging on the brain′s walking network and its mechanism.Methods:Twenty healthy elderly people and 22 healthy young adults formed an elderly group and a youth group. All were evaluated using the Mini-Mental State Examination (MMSE), the Timed Up and Go test (TUGT), the 10-metre walk test (10MWT), the functional near infrared spectroscopy walking synchrony test and GaitRite gait parameters. The intensity of functional connections and the gait parameters of the prefrontal cortex (PFC) and the primary motor cortex (MC) were compared between the two groups.Results:Compared with the youth group, the average cadence of the elderly group was significantly faster. The FC value of the RPFC in the homologous ROI, as well as those of the RMC-RPFC and RPFC-LPFC in the heterologous ROI of the elderly group were significantly lower than in the youth group.Conclusions:Lower FC values in the RPFC and its associated brain regions in the elderly during normal walking may be what activates the brain′s walking network in the early stage of brain aging.

Objective To investigate the effect of the number of previous spontaneous abortions on the first in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) cycle. Methods A retrospective case?control study was conducted to analyze the clinical data of 1 279 patients who received IVF/ICSI treatment for the first time from July 2014 to July 2018 in Tianjin Central Hospital of Gynecology Obstetrics;they were divided into 0 time group (group A, n=924), 1 time group (group B, n=267) and 2 times group (group C, n=88) for comparison, according to the previous frequency of spontaneous abortions. Results There were no statistically significant differences in age, basal testosterone, estradiol, progesterone, prolactin and embryo quality in group A, B and C (all P>0.05). The biochemical pregnancy rate of group C (9.1%) was higher than those of the other two groups (4.1% and 4.1%; all P>0.05). The clinical pregnancy rate of group A (42.5%) [>group B (40.4%) and>group C (35.2%)] was not statistically significant(P>0.05).Early abortion rate in group A (8.9%) was0.05). Conclusions Women with a history of one? or two?time spontaneous abortion have no obvious effect on embryo quality, and have a negative impact of clinical pregnancy rate, early abortion rate, live birth rate in the first IVF/ICSI cycle; especially for patients with a history of two times spontaneous abortion, early abortion rate has a significant increase, live birth rate decreases significantly, suggesting that patients with a history of two times of spontaneous abortion should find the causes of miscarriage according to the process of recurrent spontaneous miscarriage and receive treatment if necessary before subsequent pregnancy.

Objective: To investigate the effect of the number of previous spontaneous abortions on the first in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) cycle. Methods: A retrospective case-control study was conducted to analyze the clinical data of 1 279 patients who received IVF/ICSI treatment for the first time from July 2014 to July 2018 in Tianjin Central Hospital of Gynecology Obstetrics;they were divided into 0 time group (group A, n=924), 1 time group (group B, n=267) and 2 times group (group C, n=88) for comparison, according to the previous frequency of spontaneous abortions. Results: There were no statistically significant differences in age, basal testosterone, estradiol, progesterone, prolactin and embryo quality in group A, B and C (all P>0.05). The biochemical pregnancy rate of group C (9.1%) was higher than those of the other two groups (4.1% and 4.1%; all P>0.05). The clinical pregnancy rate of group A (42.5%) [>group B (40.4%) and >group C (35.2%)] was not statistically significant(P>0.05).Early abortion rate in group A (8.9%) wasP<0.05).The live birth rate of group C (21.6%) was significantly lower than that of group A (35.5%; P<0.05), but the difference between group C and group B (30.7%) was not statistically significant (P>0.05). Conclusions Women with a history of one- or two-time spontaneous abortion have no obvious effect on embryo quality, and have a negative impact of clinical pregnancy rate, early abortion rate, live birth rate in the first IVF/ICSI cycle; especially for patients with a history of two times spontaneous abortion, early abortion rate has a significant increase, live birth rate decreases significantly, suggesting that patients with a history of two times of spontaneous abortion should find the causes of miscarriage according to the process of recurrent spontaneous miscarriage and receive treatment if necessary before subsequent pregnancy.