Objective To investigate the infection situation of L-form bacteria in urinary calculi from the patients with urolithia-sis in a hospital to provide a scientific basis for postoperative anti-infection and prevention of urinary stone recurrence.Methods The calculi samples in 265 cases of urinary calculi from October to December 2015 were collected and performed the culture of com-mon bacteria and L-form bacteria respectively.Culture of common bacteria and bacterial L-forms.Results Among 265 cases of uri-nary calculi ,8 cases(3% ,8/265) were L-form Bacterial combined with common bacterial infection ,only 7 cases(2.6% ,7/265) were L-form bacterial infection ,80 cases (30.0% ,80/265) were common bacterial infection.15 strains of L-form bacteria were detected and 96 strains of common bacteria were detected.The drug resistance of L-form bacteria was significantly increased compared with common bacteria.Conclusion The positive rate of L-form bacteria culture of urinary calculi is lower than other domestic reports. Adding hypertonic medium for conducting L-form bacterial isolation and culture in the patients with urinary tract infection can re-duce the false negative.

Objective To analyze the effects of fetal reduction in early pregnancy on obstetric and neonatal outcomes of spontaneously or selectively reduced multiple pregnancies produced by in vitro fertilization-embryo transfer (IVF-ET). Methods Retrospective study of 6917 clinical pregnancies from IVF-ET cycles, including 754 multiple pregnancies divided into two groups according to the remaining fetus number: reduced singleton group (n=599) and reduced twin group (n=155); and maternal and neonatal outcomes of two groups were compared to primary singleton group (n=3589) and primary twin group (n=2574). Results The rate of pregnancy complication [9.85%(59/599) versus 6.21%(223/3589)], preterm birth [19.37%(116/599) versus 10.73%(385/3589)], low birth weight [9.71%(56/577) versus 4.57%(152/3324)], perinatal death [0.69%(4/577) versus 0.12%(4/3324)] and malformation [2.95%(17/577) versus 1.02%(34/3324)] in reduced singleton group were significantly higher than those in primary singleton group (all P<0.01). There were no significant differences between reduced twin group and primary twin group (all P>0.05). In reduced singleton group, birth defect rate was 2.95%, which was higher than those of the other three groups (P<0.05), in this group spontaneous pregnancy reduction accounted for 89.3%(535/599). Conclusions (1) The rate of pregnancy complication, preterm birth, low birth weight, perinatal death and malformation in reduced singleton group are still higher than primary singletons, suggesting embryo reduction only is a compensated method in multiple pregnancies. Limiting the number of embryos transferred is the essential solution. (2) The rate of birth defect in spontaneous pregnancy reduction group is higher, so prenatal examination should be reinforced in this group.

AIM: To investigate the cell cycle arrest ind uced by hypoxia, hypoxia inducible factor-1 and their possible mechanism in huma n ovarian cancer cell line SW626. METHODS: CoCl 2, a chemical inducer of hypoxia and hypoxic cell culture chamber were used to induce chemical and physical hypoxia in human ovar ian cancer cell line SW626. The method of ‘decoy’ was used to block the functi on of HIF-1? because it acts as the core sequence of the target gene as a compe titor combined to the HIF-1?. The cells were divided into group A1 (normal oxyg en), A2 (normal oxygen plus HIF-1? decoy), B1 (CoCl 2), B2 (CoCl 2 plus HIF-1 ? decoy), C1 (hypoxia) and C2 (hypoxia plus HIF-1?). The expression of the HIF -1? protein, mRNA and cell cycle analysis were detected by Western blotting, RT -PCR and flow cytometry (FCM). RESULTS: The expression level of HIF-1? protein in group B1 (3 .75?1.31) and group C1 (3.48?1.01) was significantly higher than that in g roup A1 (0.97?0.31) (P0.05). FCM showed that the G 0/ G 1 phase was markedly increased in group B1 (81.78?24.33) and group C1 (77 .62?22.76) and was significantly higher than that in group A1 (49.49?18.54 ) (P0.05). CONCLUSION: Both CoCl 2 and physical hypoxia could distinctly i nduce cell cycle arrest in G 0/G 1 phase and the expression of HIF-1? in huma n ovarian cancer cell line SW626. HIF-1? plays an important role in cell cycle arrest induced by hypoxia in human ovarian cancer cell line SW626.

Objective To establish an aging model of rat bone marrow stromal cells (BMSCs) in vitro and in vivo, in order to study the senescence biology of aging BMSCs .Methods The control cell group ( in vitro):isolating, puri-fying and culturing BMSCs from healthy male SD rats .collecting the third generation ( P3) of BMSCs for analysis . The aging model group (in vitro):the P3 BMSCs were incubated with D-Galactose (D-Gal, 30 g/L) for 48 hours. The aging rat model group ( in vivo): the rats were given 120 mg D-Gal by the way of daily neck subcutaneous injection for 42 consecutive days .The control rat group ( in vivo):the rats were administrated with the same volume of saline for the same times .On the second day after the aging model was established , the BMSCs were collecting and culturing for study.1)The proliferative potency was detected by cell counting Kit-8(CCK-8);the distribution of cell cycle and apoptosis was detected by flow cytometry (FCM);2)the ratio of aging BMSCs was examined by the senescence-associated β-Galactosidase(SA-β-Gal) staining;3)malonaldehyde(MDA) content and total super-oxide dismutase(SOD) was examined activity by enzymatic assay; the level of reactive oxygen species (ROS) by DCFH-DA fluorescent staining was counted with FCM;4 ) the expression level of senescence-related signaling was proteins of P16 , P21 , P53 , CDK2 and cyclin D by Western blot .Results Compared with the matched control group, the BMSCs of aging model group displayed a significant decrease in proliferation; the BMSCs were held in G1 phase arrest as the proportion of the cells in G 1 phase increased , while that decreased in S phase ( P<0.05 );and the positive ratio of SA-β-Gal stained BMSCs also significantly increased ( P <0.05 ); BMSCs in the aging model group showed an increasing level of ROS and MDA , meanwhile a decline in total SOD activity was decreased (P<0.05);P16,P21 and P53 protein expression in aging BMSCs was obviously enhanced (P<0.05), at the same time the expression of CDK2 and cyclin D was also decreased ( P<0.05 ) .Conclusions D-Gal can be used to develope an aging model of BMSCs .It acts through up-regulation of expressions of aging-related proteins and in-hibition of oxidative stress injury and chronic inflammation .

Objective To explore the effective means and important significance for preventing the born of neonatal patients with severe thalassemia.Methods Among the pregnant women and spouses receiving prenatal examination in our hospital from January 2013 to December 2015 were performed the thalassemia screening and gene diagnosis,49 couples carrying the same type thalassemia were conducted the prenatal amniotic fluid thalassemia gene diagnosis and follow up after prenatal diagnosis.Results In 49 couples carrying the same type thalassemia,the main gene mutation types of α-thalassemia detected by the gene diagnosis were --SEA/aα(50.0％),-α3.7/αa (36.5％) and-α4.2/αa (11.5％),which of β-thalassemia were CD17/N(42.0％),CD41-42/N (26.0％) and IVS-Ⅱ-654/N(22.0％).The results of prenatal diagnosis showed that there were 4 cases of HbH disease,2 cases of Bart's hydrops fetus,10 cases of severe β-thalassemia,19 a-thalassemia carriers,10 β-thalassemia carriers,1 case of co-inheritance of a-and β-thalassemia,and 3 health fetuses.The follow up results were consistent with those of prenatal diagnosis.Conclusion Conducting prenatal screening and diagnosis of thalassemia in pregnant women can effectively prevent the birth of neonatal patients with severe thalassemia.

Objective:To investigate the impact of trigger timing of gonadotropin- releasing hormone (GnRH) antagonist regimen for infertility patients of various ages.Methods:This was a retrospective study, 1 529 infertility patients who receiving GnRH antagonist regimen in Chongqing Health Center for Women and Children from January 2017 to December 2018 were divided into the advance trigger group and the standard trigger group, and further divided into three subgroups according to age:<35 years, 35-40 years,>40 years. The number of retrieved oocytes and transplantable embryos, the clinical pregnancy rate and the live birth rate among patients in the advance trigger group and standard trigger group in various age subgroups were compared.Results:(1) The gonadotropin (Gn) days among the three age subgroups were significantly shorter in the advance trigger group compared to the same-aged standard trigger group (all P<0.01), but only in the 35－40 years and >40 years subgroups, the Gn doses in the advance trigger group [(2 702±551) and (2 780±561) U] were significantly less than those in the standard trigger group (all P<0.01). In the <35 years subgroup, the number of oocytes retrieved and transplantable embryos of the advance trigger group (6.6±4.8 and 2.6±2.7) were significantly less than those of the standard trigger group (all P<0.01), but there was no difference in the number of top-quality embryos ( P=0.580); however, in the 35－40 years and >40 years subgroups, there were no significant differences between advance and standard trigger groups in terms of the afore mentioned 3 indicators (all P>0.05), only the numbers of top-quality embryos in the advance trigger group (0.6±1.0 and 0.6±0.9) were significantly higher than those in the standard trigger group (all P<0.01). (2) In the <35 years and 35－40 years subgroups, no significant differences were noted between the advance trigger group and standard trigger group with regard to the clinical pregnancy rate and live birth rate (all P>0.05); but in the >40 years subgroup, the clinical pregnancy rate of the advance trigger group was significantly higher than that of the standard trigger group [33.0% (30/91) vs 19.2% (25/130), P=0.020], and there was no statistical difference in the live birth rate ( P=0.064). (3) Multivariate logistic regression analysis showed that trigger timing was an independent predictor of clinical pregnancy rate in the >40 years subgroup ( OR=0.334, 95% CI: 0.119-0.937, P=0.037), but not an independent predictor of live birth rate ( P>0.05). Conclusions:Advance trigger in the GnRH antagonist protocol for infertility patients >40 years old could effectively reduce Gn times and Gn dosage, increase the number of top-quality embryos, and improve the clinical pregnancy rate. Therefore, compared with patients ≤40 years of age, patients >40 years might benefit more from advance trigger.

BACKGROUND:In recent years,the demand for in vitro maturation of immature oocytes has increased.Oocyte maturation is affected by many factors,among which the selection of medium is particularly important,and there is currently no unified plan. OBJECTIVE:To compare the in vitro maturation of germinal vesicle stage oocytes with different maturation media and to investigate its effects on oocyte quality and developmental potential. METHODS:Germinal vesicle oocytes were matured in G-1TM PLUS medium,CZB medium and M16 medium,and mature oocytes in vivo were used as control group to compare in vitro fertilization and early embryo development among various groups.The immunofluorescence method was used to evaluate mitochondrial function in mature oocytes of each group.Calcium oscillation was detected by confocal microscopy real-time imaging system. RESULTS AND CONCLUSION:(1)There was no significant difference in the first polar body ejection rate among the three groups(P>0.05).(2)The rate of in vitro fertilization was higher in the G-1TM PLUS group(52.86±11.24)%than that in the M16 group(37.76±6.70)%and the CZB group(30.62±5.51)%.The blastocyst rate was lower in the CZB group(36.23±6.63)%than that in the control group(78.16±4.17)%,G-1TM PLUS group(55.75±7.63)%and M16 group(53.36±6.33)%.(3)Compared with the control group,the length-to-width ratio of the spindle in the CZB group increased(P<0.005).(4)The mitochondrial function of the CZB group was worse than that of the control group,G-1TM PLUS group and M16 group,and abnormal mitochondrial agglutination occurred in the CZB group.(5)The frequency of calcium oscillations in the CZB and M16 groups was significantly higher than that in the G1 and control groups.In conclusion,during in vitro maturation of mouse oocytes,in vitro maturation rate was not significantly different among G-1TM PLUS,CZB and M16 media,but the G-1TM PLUS medium had a higher rate of fertilization and blastocyst formation.

BACKGROUND:Superovulation is a common therapy in assisted reproductive technology.In clinical practice,some patients experience repeated superovulation to get pregnant. OBJECTIVE:To explore the effect of repeated superovulation on the developmental potential of oocytes in mice and humans. METHODS:Both animal experiments and retrospective clinical research were conducted.The animal study involved 90 SPF grade ICR 8-week-old female mice,who were randomly divided into three groups for 1,3,and 5 superovulations,respectively.The clinical study involved 306 patients who had undergone three consecutive in vitro fertilization cycles.The number of ovules obtained and embryonic development in different cycles were compared. RESULTS AND CONCLUSION:(1)The animal study indicated that repeated superovulation did not affect the embryonic development or developmental speed of mouse embryos.Similarly,there was no significant difference in the mouse blastocyst apoptosis,DNA damage,or the formation of inner cell mass and trophectoderm(P>0.05).(2)The clinical study also revealed no significant differences in the number of retrieved oocytes(8.60±5.04,8.58±4.87,and 8.38±4.63,P=0.81)and transferable embryos(2.42±1.99,2.40±1.92,and 2.64±2.00,P=0.26)over the three cycles.(3)In both the young group(<35 years)and the old group(≥35 years),the embryo quality was not affected by repeated superovulation(P>0.05).(4)These findings show that repeated superovulation does not affect the developmental potential of oocytes in mice and humans.

Objective: To investigate the clinical application value of single nucleotide polymorphism (SNP) haplotype analysis in the preimplantation genetic diagnosis (PGD) of monogenic diseases. Methods: The whole genome amplification products of biopsied trophectoderm cells were analyzed by SNP haplotype analysis and verified by Sanger sequencing. Results: A total of 205 embryos were performed SNP haplotype analysis and Sanger sequencing. Among them, Sanger sequencing failed in 14.63% (30/205) of embryos, and SNP haplotype analysis failed in 0.98% (2/205) of embryos. The failure rate of the latter was significantly lower than that of the former (P＜0.05). There were consistent results in 155 (75.61%) embryos, and inconsistent results in 18 (8.78%) embryos. Forty-five embryos in 41 cycles were performed embryo transplantation. The clinical pregnancy rate was 70.73% (29/41) and the implantation rate was 71.11% (32/45). The results of prenatal diagnosis of amniotic fluid during the second trimester of pregnancy were completely consistent with those of SNP haplotype analysis. Conclusion SNP haplotype analysis is accurate, and its failure rate is lower than that of Sanger sequencing. It can be effectively used in the PGD of clinical monogenic diseases.

The lower extremity exoskeleton robot is a wearable device designed to help people suffering from a walking disorder to regain the power of the legs and joints to achieve standing and walking functions. Compared with traditional robots that include rigid mechanisms, lower extremity exoskeleton robots with compliant characteristics can store and release energy in passive elastic elements while minimizing the reaction force due to impact, so it can improve the safety of human-robot interaction. This paper reviews the compliant characteristics of lower extremity exoskeleton robots from the aspects of compliant drive and compliant joint, and introduces the augmentation, assistive, rehabilitation lower extremity exoskeleton robots. It also prospect the future development trend of lower extremity exoskeleton robots.