Objective:To analyze the impact of intrauterine infusion of autologous peripheral blood mononuclear cells (PBMCs) and enriched platelet plasma (PRP) on pregnancy outcomes in patients with recurrent implantation failure (RIF).Methods:A total of 96 patients with repeated implantation failures who underwent frozen embryo cycles at Hunan Provincial Maternal and Child Health Care Hospital from March 2021 to June 2023 were selected and randomly divided into a control group (19 cases), PBMCs group (31 cases), and PRP group (46 cases). The control group did not receive uterine cavity infusion treatment; Intrauterine perfusion of PBMCs in the PBMCs group; The uterine cavity of the PRP group was infused with PRP. We compared the general situation, endometrial thickness on the day of conversion, endometrial thickness on the day of transplantation, embryo implantation rate, and clinical pregnancy rate among three groups.Results:There was no statistically significant difference in age, body mass index (BMI), years of infertility, menstrual cycle, serum basal follicle stimulating hormone (FSH), basal estradiol (E 2), number of transfer cycles, number of transferred embryos, and number of high-quality embryos among the three groups (all P>0.05). There was no statistically significant difference in endometrial thickness on the conversion day among the control group, PRP group, and PBMCs group (all P>0.05). The endometrial thickness on the day of transplantation in the PRP group was greater than that in the control group and PBMCs group (all P<0.05), and there was no statistically significant difference in endometrial thickness on the day of transplantation between the control group and PBMCs group (all P>0.05). The embryo implantation rate and clinical pregnancy rate of the PRP group and PBMCs group were higher than those of the control group, and the difference was statistically significant (all P<0.05). There was no statistically significant difference in embryo implantation rate and clinical pregnancy rate between the PRP group and the PBMCs group (all P>0.05). The patients did not experience any adverse reactions such as infection, abdominal pain, or vaginal bleeding during intrauterine infusion therapy. Conclusions:Infusing autologous PBMCs or PRP into the uterine cavity before re embryo transfer in RIF patients can significantly improve embryo implantation rate and clinical pregnancy rate, and can improve assisted pregnancy outcomes; Intrauterine infusion of autologous PRP has no significant advantage over PBMCs in improving clinical pregnancy outcomes in patients with RIF; But it is more beneficial for improving the thickness of the endometrium.

Objective:To evaluate the changes of cortisol and sIgA in serum and saliva of rats under different stresses and pressures, so as to provide a basis for screening non-invasive stress monitoring indicators in high-pressure working environment.Methods:A total of 54 rats were divided into six groups exposed to different air pressures of 0 kPa, 175 kPa, 350 kPa, 500 kPa, 600 kPa, and 700 kPa, respectively. According to different stress conditions, another 36 rats were divided into physiological stress group (PSSG), psychological stress group (PCSG), physiological and psychological stress group (PPSG), and blank group (BG), with 9 rats in each group. At the end of the experiment, serum sample was collected and supernatant was taken from the sublingual gland tissue homogenate. Cortisol and sIgA levels in saliva and serum were determined by radioimmunoassay and enzyme-linked immunosorbent assay (ELISA).Results:The concentrations of SIgA in group 500 kPa, 600 kPa, and 700 kPa were significantly different ( F=10.961, P<0.001; F=4.693, P=0.001; F=4.353, P=0.003). The serum cortisol levels in group 175 kPa, 350 kPa, and 600 kPa were significantly higher than that in group 0 kPa, while the serum cortisol level in group 350 kPa was significantly higher than those in group 500 kPa and 700 kPa. The cortisol concentrations of sublingual gland tissues in group 350 kPa and 700 kPa were significantly higher than those in group 0 kPa, 500 kPa, and 600 kPa. Under different stress conditions, serum sIgA concentrations in PCSG, PSSG, and PPSG were significantly lower than that in BG ( F=4.852, P=0.007; F=4.918, P=0.007; F=3.967, P=0.017). The levels of serum and sublingual cortisol in PCSG and PPSG were significantly higher than those in BG, while the levels of serum and sublingual cortisol in PSSG were significantly lower than those in PCSG. Relevant research results showed that the changes of salivary gland cortisol and serum cortisol were positively correlated under different stress conditions ( r=0.609, P<0.01). Conclusion:Cortisol as a stress monitoring indicator has good effectiveness, and using saliva as an indicator to monitor stress under high pressure environment has a certain degree of feasibility.

Objective:To evaluate the changes of cortisol and sIgA in serum and saliva of rats under different stresses and pressures, so as to provide a basis for screening non-invasive stress monitoring indicators in high-pressure working environment.Methods:A total of 54 rats were divided into six groups exposed to different air pressures of 0 kPa, 175 kPa, 350 kPa, 500 kPa, 600 kPa, and 700 kPa, respectively. According to different stress conditions, another 36 rats were divided into physiological stress group (PSSG), psychological stress group (PCSG), physiological and psychological stress group (PPSG), and blank group (BG), with 9 rats in each group. At the end of the experiment, serum sample was collected and supernatant was taken from the sublingual gland tissue homogenate. Cortisol and sIgA levels in saliva and serum were determined by radioimmunoassay and enzyme-linked immunosorbent assay (ELISA).Results:The concentrations of SIgA in group 500 kPa, 600 kPa, and 700 kPa were significantly different ( F=10.961, P<0.001; F=4.693, P=0.001; F=4.353, P=0.003). The serum cortisol levels in group 175 kPa, 350 kPa, and 600 kPa were significantly higher than that in group 0 kPa, while the serum cortisol level in group 350 kPa was significantly higher than those in group 500 kPa and 700 kPa. The cortisol concentrations of sublingual gland tissues in group 350 kPa and 700 kPa were significantly higher than those in group 0 kPa, 500 kPa, and 600 kPa. Under different stress conditions, serum sIgA concentrations in PCSG, PSSG, and PPSG were significantly lower than that in BG ( F=4.852, P=0.007; F=4.918, P=0.007; F=3.967, P=0.017). The levels of serum and sublingual cortisol in PCSG and PPSG were significantly higher than those in BG, while the levels of serum and sublingual cortisol in PSSG were significantly lower than those in PCSG. Relevant research results showed that the changes of salivary gland cortisol and serum cortisol were positively correlated under different stress conditions ( r=0.609, P<0.01). Conclusion:Cortisol as a stress monitoring indicator has good effectiveness, and using saliva as an indicator to monitor stress under high pressure environment has a certain degree of feasibility.

Objective:To evaluate the effects of different high pressure on the behavior and physiological indexes of hypothalamic-pituitary-adrenal axis (HPA) in rats, so as to provide a theoretical basis for the stress protection in decompression process.Methods:The rats were divided into four groups according to the random number table method and were separately exposed to air pressure of 0 kPa, 170 kPa, 350 kPa and 700 kPa for 60 mins and given decompression for 45 mins. The behaviors of rats were analyzed by animal open field method and video analysis system of the open field immediately after they were taken from the air pressure chamber, and the contents of adrenocorticotrophic hormone (ACTH) and glucocorticoid (GC) in the serum were measured by the enzyme-linked immunosorbent assay (ELISA).Results:Under different pressure, compared with the 0 kPa group, the total distance traveled by the rats ( F=3.354), the average traveling speed ( F=3.358), the number of standing times and standing time duration ( F=3.739; F=33.332) of the rats in the other 3 groups decreased significantly. The distance traveled in the corner of the 175 kPa group and the 700 kPa group decreased significantly ( F=3.532), while their cleaning time significantly increased ( F=4.581). The standing times and the cleaning time of the 350 kPa group were significantly less than those of the 700 kPa group. All the differences above showed statistical significance ( P<0.05). After exposure to different pressure, the ACTH level of the 175 kPa group was higher than those of the 0 kPa group and the 350 kPa group ( F=5.309) with statistical significance ( P<0.05). The GC level of the 0 kPa group was significantly lower than those of the other 3 groups, and the GC level of the 700 kPa group was especially lower than that of the 175 kPa group, with statistically significant difference ( P<0.05). Conclusion:High pressure exposure at all levels can cause definite stress reaction, and the stress reactions do not correlate with the pressure gradient. Within the physiological bearable range of stress under high pressure, the individual stress level may present a certain steady state. The individual′s behavioral response, ACTH and GC levels may be used as indicators for stress monitoring.

Objective:To evaluate the effects of different high pressure on the behavior and physiological indexes of hypothalamic-pituitary-adrenal axis (HPA) in rats, so as to provide a theoretical basis for the stress protection in decompression process.Methods:The rats were divided into four groups according to the random number table method and were separately exposed to air pressure of 0 kPa, 170 kPa, 350 kPa and 700 kPa for 60 mins and given decompression for 45 mins. The behaviors of rats were analyzed by animal open field method and video analysis system of the open field immediately after they were taken from the air pressure chamber, and the contents of adrenocorticotrophic hormone (ACTH) and glucocorticoid (GC) in the serum were measured by the enzyme-linked immunosorbent assay (ELISA).Results:Under different pressure, compared with the 0 kPa group, the total distance traveled by the rats ( F=3.354), the average traveling speed ( F=3.358), the number of standing times and standing time duration ( F=3.739; F=33.332) of the rats in the other 3 groups decreased significantly. The distance traveled in the corner of the 175 kPa group and the 700 kPa group decreased significantly ( F=3.532), while their cleaning time significantly increased ( F=4.581). The standing times and the cleaning time of the 350 kPa group were significantly less than those of the 700 kPa group. All the differences above showed statistical significance ( P<0.05). After exposure to different pressure, the ACTH level of the 175 kPa group was higher than those of the 0 kPa group and the 350 kPa group ( F=5.309) with statistical significance ( P<0.05). The GC level of the 0 kPa group was significantly lower than those of the other 3 groups, and the GC level of the 700 kPa group was especially lower than that of the 175 kPa group, with statistically significant difference ( P<0.05). Conclusion:High pressure exposure at all levels can cause definite stress reaction, and the stress reactions do not correlate with the pressure gradient. Within the physiological bearable range of stress under high pressure, the individual stress level may present a certain steady state. The individual′s behavioral response, ACTH and GC levels may be used as indicators for stress monitoring.

Objective To explore causes and solutions of false estimates of elevated serum creatinine in patients with Walden‐strom′s macroglobulinemia throug wet chemical enzymatic method .Methods 5 cases of patients hospitalized in the Bethune Inter‐national Peace Hospital were enrolled as subjects from 2010 to 2012 .The large molecular proteins were removal from serum sam‐ples collected from patients with Waldenstrom′s macroglobulinemia by using centrifugal ultrafiltration tube .The serum creatinine levels were detected through using the wet chemical enzymatic method ,wet chemical picric acid method and dry chemical enzymatic method before and after ultrafiltration ,and data were compared .Results Before ultrafiltration ,the levels of serum creatinine of 2 cases of patients with Waldenstrom′s macroglobulinemia detected by using wet chemical enzymatic method differed with those de‐tected by using wet chemical picric acid method and dry chemical enzymatic method .While there were no obvious differences be‐tween levels of serum creatinine detected by wet chemical picric acid method and dry chemical enzymatic method .While ,after ultra‐filtration ,no obvious differences were founded in levels of serum creatinine detected by the thress methods .Conclusion The large molecular proteins should be eliminated when using the wet chemical enzymatic method in the detection of serum creatinine levels , in order to avoid abnormal increase .And the wet chemical picric acid method and dry chemical enzymatic method could also be uti‐lized to determine the accuracy ,and provide reliable determination results .

Objective To clone and express the Tp0453 antigen immuno-dominant epitope fragment of Treponema pallidum (Tp) in Escherichia coli,in an effort to develop serological tests with increased specificity for the diagnosis of syphilis.Methods The gene encoding Tp0453 recombinant outer membrane protein fragment was amplified by polymerase chain reaction (PCR),and inserted into expression vector pQE30 after T-A cloning,then confirmed by restriction map.The constructed recombinant plasmid pQE30-Tp0453 was transformed to E.coli M15 for expression induced by isopropyl β-D-1-thiogalactopyranoside.The expressed product was identified by Western blot,and purified by Ni2+-NTA agarose column chromatography.A double antigen sandwich enzymelinked immunosorbent assays (ELISA) was established by using the recombinant Tp0453 protein to test sera from 48 patients with positive Treponema pallidum particle agglutination test (TPPA),and 40 negative sera as control.Results The PCR amplicon of the target gene was about 490 bp.The recombinant plasmid pQE30-Tp0453 was correctly constructed and successfully expressed in E.coli M15.The expressed product,with a relative molecular of about 21 000,existed in a form of inclusion body,accounting for about 18％ of total somatic protein,and reached a purity of more than 95％ after purification.Western blot showed specific reaction of the expressed protein with Tp positive serum.The ELISA tests with the 88 clinical samples yielded a sensitivity of 97.9％ (47/48),and specificity of 100.0 ％ (40/40).The consistency of results between the ELISA test and the TPPA test was 98.9 ％ (87/88).Conclusion The expressed Tp0453 fragment has showed good immunoreactivity with serum from patients with syphilis,providing the foundation of further development of serological diagnostic kit with increased specificity for the diagnosis of TP infection.

A 31-year-old male normotensive patient with aldosterone-producmg adenoma complained of thirst,polydipsia,polyuria,and periodical paraplegia.The diagnosis is raised by signs of hypokalemia.Despite the lack of hypertension,primary aldosteronism was confirmed by persistent hypokalemia,increased urinary potassium,increased urinary and plasma aldosterone levels and suppressed plasma rennin activity (PRA).The blood pressure profile was studied by ambulatory monitoring,and the mean blood pressure of 24h was normal and the circadian rhythm remained normal. Surgical removal of the histologically typical aldosterone-producing adenomas normalized the kalemia.The patient had a marked fall in blood pressure with mean values of 21/17 mm Hg ( diurnal and nocturnal blood pressure were 19/17 and 22/17 mm Hg respectively)and recovery of normal urinary and plasma aldosterone levels and PRA 6 weeks after surgery.This suggests that excess serum aldosterone induced relative hypertension in those patients whose blood pressure was spontaneously very low.Our observations call for primary hyperaldosteronism assay in patients with hypokalemia and renal potassium leakage.

Objective To investigate the effects of propofol on the spontaneous transient outward K+ currents in mouse cerebral arterial smooth muscle cells.Methods Kunming mice of both sexes,weighing 18-22 g,were used in this study.Vascular smooth muscle cells were freshly isolated from cerebral arteries in two steps.Five cells were chosen and studied.When holding potential was - 30 mV,spontaneous transient outward K+ currents were recorded before and after the application of 56 μmol/L propofol by perforated whole-cell patch-clamp technique.The amplitude,frequency,area under the curve and half time width of spontaneous transient outward K+ currents were analyzed.Results Propofol 56 μmol/L significantly increased the amplitude,frequency and area under the curve of spontaneous transient outward K+ currents.There was no significant change in the half time width of spontaneous transient outward K+ currents after administration,Conclusion Propofol can activate spontaneous transient outward K+ currents in mouse cerebral arterial smooth muscle cells,and thus induces vascular smooth muscle relaxation.

Objective To investigate the effects of intestinal ischemia-reperfusion(I/R)on cerebral microglial activation in rats.Methods One hundred and twenty-eight healthy male SD rats weighing 250-300 g were randomly allocated to one of two groups(n =64 each):group sham operation(group S)and intestinal I/R group.Intestinal I/R was produced by occlusion of superior mesenteric stery for 90 main followed by reperfusion.Sixteen animals were sacrificed at each of the 4 time points:2,6,24 and 48 h of reperfusion in each group.Their intestines were obtained for microscopic examination.Their brains were harvested for detection of microglial activation (by immuno-histochemistry).The reactive oxygen species(ROS),MDA and NO contents and SOD,nitric oxide synthase(NOS)and inducible nitric oxide synthase(iNOS)activities in the brain were measured.Results The microglia were in quiescent condition.Ibal staining was negative or light in group S.Intestinal I/R significantly increased intestinal Chiu score,cerebral microglial activation at 6,24 and 48 h of repeffusion which peaked at 24 h of reperfusion in group I/R as compared with group S.Cerebral ROS,MDA,NO levels and NOS,iNOS activities were significantly higher while SOD activity was significantly lower in group I/R than in group S.Concluslon Intestinal I/R can activate microglia and induce the release of nitrogen and oxygen free radicals resulting in cerebral injury.