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 explore the correlation between blastomere count variations “skip value” which extracted from by time-lapse technology (TLT) combined with artificial intelligence (AI) and morphological features of in vitro fertilization (IVF) embryo, and to test its feasibility in clinical applications.Methods:This study was a diagnostic experiment (AI reassessment of embryo transferred patients), a total of 6 545 embryos from 1 226 patients who underwent IVF at the Women and Children′s Hospital of Chongqing Medical University from December 2020 to December 2021 were retrospectively analyzed, of which 2 869 embryos were attempted to cultured to blastocyst stage by TLT. The embryo dynamic map (EDM) was drawn by Embryo Viewer, a TLT recording software, based on embryo developmental kinetics. The self-developed AI embryo evaluation software identified and recorded the number of cleavages in real time during embryonic development, and compared with the EDM, the correlation between the skip value formed by the change of cleavage sphere counts and the outcomes of the embryos was analyzed. The correlation among skip value, morphological score of embryo, implantation rate and live birth rate were performed by Spearman and step-up logistic regression. The receiver operating characteristic (ROC) curve was selected for reporting there relationship of skip value and morphology. Finally, predicting power of skip value for implantation and live birth rate were performed by ROC analysis.Results:The total skip values extracted from the blastomere count of embryos (72 hours post-fertilization) were negatively correlated with abnormal cleavage, blastocyst formation rate, day 3 (D3)-cell score, uneven size and fragmentation (the β values were -0.268, -0.116, -0.213, -0.159 and -0.222, respectively; all P<0.001); positively correlated with D3-cell number ( β=0.034; P<0.001); negatively correlated with blastocyst formation rate and implantation rate ( OR=0.97, 95% CI: 0.93-0.99, P=0.034; OR=0.96, 95% CI: 0.93-0.98, P=0.044). The power of predicting implantation were similar between the order selection of skip values and traditional morphology criteria [area under curve (AUC): 0.679 vs 0.620]. Live birth rate were negatively correlated with female age ( OR=0.91, 95% CI: 0.88-0.93; P<0.001), D3 general score ( OR=0.77, 95% CI: 0.59-0.99; P=0.045) and order selection of skip values ( OR=0.98, 95% CI: 0.96-0.99; P=0.038), while positively correlated with retrieved oocyte number and endometrial thickness in embryo transferred ( OR=1.08, 95% CI:1.05-1.11, P<0.001; OR=1.09, 95% CI:1.06-0.12, P<0.001, respectively) from multivariate regression analysis, and the power of predicting live birth was 0.666 for AUC. Conclusions:The skip value and its order form is a systematic quantification of embryo development, correlated with embryo developmental quality and clinical outcome. It could be an addition parameter for embryo culture and selection.

Objective:To investigate the knowledge of Sixth Chinese national consensus report on the management of Helicobacter pylori infection ( treatment excluded) (hereinafter referred to as sixth national consensus) and 2022 Chinese national clinical practice guideline on Helicobacter pylori eradication treatment (hereinafter referred to as the guideline)among medical staff from general hospitals in Hainan. Methods:From February 20 to May 7, 2023, a questionnaire survey on the diagnosis and treatment of Helicobacter pylori ( H. pylori) infection was conducted among 1 463 medical staff from 15 general hospitals in Hainan Province. The questionnaire was drawn up according to the sixth national consensus and the guideline, covering knowledge of 6 sections, induding H. pylori related diseases, detection of H. pylori, eradication, prevention and influence factors of eradication of H. pylori, etc. Chi-square test was used for statistical analysis. Results:A total of 1 463 valid questionnaires were collected with the effective responsive rate of 100.00%.The 1 463 subjects included 225 gastroenterologists and 1 238 other medical staff(including 503 physicians from other departments, 264 surgeons and 471 medical technologists and pharmacists). About 78.67%(177/225)of gastroenterologists agreed that the overall infection rate of H. pylori in China was more than 20%, the awareness rate was higher than that of other medical staff (physicians from other departments 65.41%(329/503), surgeons 61.74%(163/264), medical technologists and pharmacists 60.30%(284/471); the following datas were sorted by this position), and the difference was statistically significant ( χ2=30.97, P<0.001). About 51.11%(115/225) of gastroenterologists considered that H. pylori serological antibody test could not be used as a diagnostic method for current infection, the awareness rate was higher than that of other medical staff(22.07%(111/503), 14.02%(37/264), 12.31%(58/471)), and the difference was statistically significant( χ2 =152.66, P<0.001). Proton pump inhibitor and potassium-competitive acid blocker should be discontinued for 2 weeks, and antibiotics and bismuth should be discontinued for 4 weeks before urea breath test, and the awareness rates of gastroenterologists were higher than those of other medical staff (38.67%(87/225) vs. 23.26%(117/503), 19.70%(52/264), 18.47%(87/471); 60.89%(137/225) vs. 26.64%(134/503), 25.76%(68/264), 23.78%(112/471)), and the differences were statistically significant ( χ2 =133.70 and 165.51, both P<0.001). For refractory H. pylori infection, 98.67%(222/225)of gastroenterologists agreed with the individualized diagnosis and treatment of H. pylori infection should be guided by bacterial culture, antibiotic susceptibility test or drug resistance gene test, and the awareness rate was higher than that of other medical staff (91.85%(462/503), 93.56%(247/264), 93.21%(439/471)), and the difference was statistically significant( χ2=20.55, P=0.002). About 70.67% (159/225) of gastroenterologists recommended a bismuth containing quadruple regimen, 80.44% (181/225) supported a 10 to 14 day H. pylori eradication course, and the awareness rates were higher than other medical staff (46.92%(236/503), 33.33%(88/264), 32.91%(155/471); 67.20%(338/503), 59.09%(156/264), 53.93%(254/471)), and the differences were statistically significant ( χ2=111.25 and 59.99, both P<0.001). The understanding rates of the sixth national consensus and the guideline in gastroenterologists was 85.33% (192/225), which was higher than that of other medical staff (64.21%(323/503), 66.67%(176/264), 57.96%(273/471)), and the difference was statistically significant ( χ2=85.47, P<0.001). Conclusions:Gastroenterologists from general hospitals in Hainan Province have a better understanding of the sixth national consensus and the guideline than other medical staff. However, there is still a lack of deep understanding of the sixth national consensus and the guideline, and it is necessary to further strengthen the learning and application of the sixth national consensus and the guideline.

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.

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.

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 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.