Type 2 inflammation（T2 inflammation）is an immune response mediated by Th2 cells，eosinophils，type 2 innate lymphocytes，and other cells. Its pathological characteristics are manifested by the abnormally high expression of type 2 cytokines such as IL-4，IL-5，and IL-13，and it widely involves allergic diseases including asthma，allergic rhinitis，and atopic dermatitis. Ginsenosides are the core active ingredients of ginseng and the key substances for ginseng to exert its pharmacological effects. It has anti-inflammatory and immunomodulatory effects. This article reviews the mechanism of action，experimental research progress，and clinical application prospects of ginsenosides in T2 inflammation，explores the therapeutic potential and clinical challenges of ginsenosides，and proposes future research directions.

Medical microbiology experiment is faced with many problems in online teaching. This study adopts the teaching mode of online live broadcast + operation video + virtual experiment, and make up the operation gap to some extent through operation video and virtual experiment. The mode of assessment is subjective thinking question (closely following the operation process) + experiment design + literature review (focusing on the key technology or new technology of clinical assessment that cannot be carried out due to the limitation of conditions in traditional experiments, such as mass spectrometry, fluorescence quantitative PCR, and G-test), and it is helpful to understand students' mastery of teaching objectives, and the ability of comprehensive application and innovative thinking. The student questionnaire shows that most students hold a positive attitude towards the online experimental teaching mode, and the quality of students' homework shows that most students have a good learning effect.

Objective:To improve the technique of oocyte retrieval in in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) cycles and reduce the cancellation rate of IVF/ICSI cycle. Methods:The clinical data of a case of IVF/ICSI cycle using transvaginal combined with abdominal oocyte retrieval was reported.Results:The right ovary of one infertile patient who received ICSI due to male factors was not detected repeatedly by transvaginal ultrasound during ovulation induction. The right lower abdominal pain occurred day 5 after using gonadotropin. We considered that the position of the right ovary was high due to pelvic adhesion after previous appendectomy, and was further confirmed by abdominal ultrasound. Two mature oocytes were obtained from the left and right ovaries respectively. Four embryos were obtained after ICSI insemination, and biochemical pregnancy occurred after transplantation.Conclusion:Transabdominal ultrasound-guided oocyte retrieval is relatively safe and effective, and can obtain similar clinical outcomes as conventional transvaginal ultrasound-guided oocyte retrieval. For IVF/ICSI women with difficulty in obtaining oocytes through vagina, transabdominal ultrasound-guided oocyte retrieval can be tried. During IVF/ICSI treatment cycles, it is necessary to improve the quality of medical history inquiry, pre-cycle evaluation, ovulation induction monitoring, oocyte retrieval and embryo transfer.

Objective:To improve the technique of oocyte retrieval in in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) cycles and reduce the cancellation rate of IVF/ICSI cycle. Methods:The clinical data of a case of IVF/ICSI cycle using transvaginal combined with abdominal oocyte retrieval was reported.Results:The right ovary of one infertile patient who received ICSI due to male factors was not detected repeatedly by transvaginal ultrasound during ovulation induction. The right lower abdominal pain occurred day 5 after using gonadotropin. We considered that the position of the right ovary was high due to pelvic adhesion after previous appendectomy, and was further confirmed by abdominal ultrasound. Two mature oocytes were obtained from the left and right ovaries respectively. Four embryos were obtained after ICSI insemination, and biochemical pregnancy occurred after transplantation.Conclusion:Transabdominal ultrasound-guided oocyte retrieval is relatively safe and effective, and can obtain similar clinical outcomes as conventional transvaginal ultrasound-guided oocyte retrieval. For IVF/ICSI women with difficulty in obtaining oocytes through vagina, transabdominal ultrasound-guided oocyte retrieval can be tried. During IVF/ICSI treatment cycles, it is necessary to improve the quality of medical history inquiry, pre-cycle evaluation, ovulation induction monitoring, oocyte retrieval and embryo transfer.

The evolution, structure and antigenic epitopes prediction of Rana dybowskii antimicrobial peptide dybowskin-1ST were carried out using bioinformatics software available online. Its antibacterial mechanism and structural properties were analyzed, and its activity was verified by applying wound healing assay in mice and bacteriostatic assay in vitro. This provides the theoretical basis for the improvement of parental peptide and the development of novel derivative peptides. The software MEGA_X were used to conduct homology alignment and to construct a phylogenetic tree. The online software ProtParam, ProtScale, PeptideCutter, signal, TMHMM Server were respectively used to predict the physicochemical parameters, hydrophilia/hydrophobicity, shear sites, signal peptides, and transmembrane domains of dybowskin-1ST. The online software SOPMA, Jpred4, DNAstar Protean were used to predict the secondary structure of dybowskin-1ST, and SWISS-MODEL, I-TASSER were used to predict the tertiary structure. ABCpred and SYFPEITHI were respectively used to predict its B-and T-cell epitopes. The effect of dybowskin-1ST on the wound healing was observed on experimental mice. Kirby-Bauer method and dilution method were used to determine the bacteriostatic activity of dybowskin-1ST. The dybowskin-1ST consists of 59 amino acid residues, of which leucine accounts for 16.9%, with a molecular formula of C₃₁₈H₅₁₀N₈₀O₉₃S₂. Its theoretical isoelectric point is 5.10 and the charge is -2. The dybowskin-1ST and dybowskin-1CDYa are closely related phylogenetically. The secondary structure of dybowskin-1ST predicted by the three methods were similar, which consisted of α-helix (44.07%), extended strand (16.95%), β-turns (3.39%), and random coil (35.39%). The prediction of tertiary structure showed that dybowskin-1ST was mainly composed of α-helix, and it was regarded as a hydrophilic protein with signal peptide sequence. Subcellular localization analysis showed that the probability of secreting the mitochondrial targeted peptides was 0.944. Dybowskin-1ST is an extracellular protein with no transmembrane structure region, but contains seven phosphorylation sites, three T-cell epitopes and eight B-cell epitopes. The dybowskin-1ST promoted wound healing and effectively inhibited the growth of Escherichia coli and Staphylococcus aureus. However, it had limited antibacterial activity against fungi and drug-resistant bacteria. Although the structure of dybowskin-1ST is rich in α-helix, the verification experiments showed that its antibacterial ability needs to be enhanced. The reason may be that it is a negatively charged and hydrophilic protein, and amino acid modification with the aim of increasing the number of positive charges and changing the hydrophobicity may be used to obtain derived peptides with enhanced activity.
Amino Acid Sequence ; Animals ; Mice ; Phylogeny ; Pore Forming Cytotoxic Proteins ; Protein Structure, Secondary ; Ranidae

Objective:To investigate the clinical effects and pregnancy outcomes of the early follicular long-term protocol and antagonist protocol in the treatment of unexplained infertility patients.Methods:From January 2018 to January 2019, 642 cases of unexplained infertility patients with early follicular long-term protocol and antagonist protocol in in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) were collected using the clinical assisted reproductive technologies management system software database of the Department of Reproductive Medicine of the First Affiliated Hospital of Nanjing Medical University. Patients were divided into early follicular long-term protocol ( n=283) and antagonist protocol ( n=359) groups. The clinical outcomes of the two groups were analyzed retrospectively, including the implantation rate, the clinical pregnancy rate, and the live birth rate. Furthermore, the pregnancy outcomes of different age and body mass index (BMI) patients were further analyzed. Results:1) The BMI in the antagonist protocol group was higher than that in early follicular long-term protocol group [(22.17±2.96) kg/m 2vs. (21.68±2.29) kg/m 2, P=0.018]. The other based data did not exhibit remarkable difference between the two groups ( P>0.05). 2) The starting dosage of gonadotropin (Gn) in early follicular long-term protocol group was less than that of antagonist group [(149.74±36.24) IU vs. (177.97±38.85) IU, P<0.001]. While the total duration and dosage of Gn used in early follicular long-term protocol group were significantly higher than those in antagonist group [(11.93±2.26) d vs. (8.86±1.45) d, P<0.001; (1 908.35±632.36) IU vs. (1 638.57±497.23) IU, P<0.001). The cleavage embryo implantation rate, the clinical pregnancy rate and the live pregnancy rate in fresh cycle in early follicular long-term protocol group were significantly higher than those in the antagonist group [57.14% (152/266) vs. 39.53% (68/172), P<0.001; 66.48% (121/182) vs. 51.72% (60/116), P=0.011; 59.89% (109/182) vs. 40.52% (47/116) , P=0.001]. The incidence of moderate and severe ovarian hyperstimulation syndrome between the two groups were not statistically different ( P>0.05). Multivariate logistic regression analysis showed that different protocols and age were both risk factors for clinical pregnancy and live birth of cleavage embryo transfer [2.261(95% CI=1.333-3.836), P=0.002; 0.928(95% CI=0.869-0.991), P=0.026; 2.598(95% CI=1.535-4.397), P<0.001; 0.906(95% CI=0.849-0.967), P=0.003]. 3) In patients under 35 years old, the clinical pregnancy rate and the live birth rate in early follicular long-term protocol group were higher than those of the antagonist group, but there was no statistical difference (all P>0.05). The fresh embryo cycle implantation rate, the clinical pregnancy rate and the live birth rate of 35-39 years old patients in the early follicular long-term protocol group were higher than those in the antagonist group, but there was no statistical difference (all P>0.05). 4) In normal BMI group, the implantation rate, the clinical pregnancy rate and the live birth rate in early follicular long-term protocol group were significantly higher than those in the antagonist group [55.71% (122/219) vs. 37.82% (45/119), P=0.002; 63.58% (96/151) vs. 46.99% (39/83), P=0.014; 58.94% (89/151) vs. 39.76% (33/83), P=0.005]. In the overweight population, the implantation rate and the clinical pregnancy rate in early follicular long-term protocol group were significantly higher than those in the antagonist group [68.09% (32/47) vs. 43.40% (23/53), P=0.013; 81.25% (26/32) vs. 57.14% (20/35), P=0.034], but there was no statistically significant difference in the live birth rate ( P>0.05). Conclusion:Compared with the antagonist protocol, early follicular long-term protocol for unexplained infertility patients may achieve higher clinical pregnancy outcomes in IVF fresh cycle, but it could increased the duration of descending, the duration and dosage of Gn used.

Objective:To investigate the clinical effects and pregnancy outcomes of the early follicular long-term protocol and antagonist protocol in the treatment of unexplained infertility patients.Methods:From January 2018 to January 2019, 642 cases of unexplained infertility patients with early follicular long-term protocol and antagonist protocol in in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) were collected using the clinical assisted reproductive technologies management system software database of the Department of Reproductive Medicine of the First Affiliated Hospital of Nanjing Medical University. Patients were divided into early follicular long-term protocol ( n=283) and antagonist protocol ( n=359) groups. The clinical outcomes of the two groups were analyzed retrospectively, including the implantation rate, the clinical pregnancy rate, and the live birth rate. Furthermore, the pregnancy outcomes of different age and body mass index (BMI) patients were further analyzed. Results:1) The BMI in the antagonist protocol group was higher than that in early follicular long-term protocol group [(22.17±2.96) kg/m 2vs. (21.68±2.29) kg/m 2, P=0.018]. The other based data did not exhibit remarkable difference between the two groups ( P>0.05). 2) The starting dosage of gonadotropin (Gn) in early follicular long-term protocol group was less than that of antagonist group [(149.74±36.24) IU vs. (177.97±38.85) IU, P<0.001]. While the total duration and dosage of Gn used in early follicular long-term protocol group were significantly higher than those in antagonist group [(11.93±2.26) d vs. (8.86±1.45) d, P<0.001; (1 908.35±632.36) IU vs. (1 638.57±497.23) IU, P<0.001). The cleavage embryo implantation rate, the clinical pregnancy rate and the live pregnancy rate in fresh cycle in early follicular long-term protocol group were significantly higher than those in the antagonist group [57.14% (152/266) vs. 39.53% (68/172), P<0.001; 66.48% (121/182) vs. 51.72% (60/116), P=0.011; 59.89% (109/182) vs. 40.52% (47/116) , P=0.001]. The incidence of moderate and severe ovarian hyperstimulation syndrome between the two groups were not statistically different ( P>0.05). Multivariate logistic regression analysis showed that different protocols and age were both risk factors for clinical pregnancy and live birth of cleavage embryo transfer [2.261(95% CI=1.333-3.836), P=0.002; 0.928(95% CI=0.869-0.991), P=0.026; 2.598(95% CI=1.535-4.397), P<0.001; 0.906(95% CI=0.849-0.967), P=0.003]. 3) In patients under 35 years old, the clinical pregnancy rate and the live birth rate in early follicular long-term protocol group were higher than those of the antagonist group, but there was no statistical difference (all P>0.05). The fresh embryo cycle implantation rate, the clinical pregnancy rate and the live birth rate of 35-39 years old patients in the early follicular long-term protocol group were higher than those in the antagonist group, but there was no statistical difference (all P>0.05). 4) In normal BMI group, the implantation rate, the clinical pregnancy rate and the live birth rate in early follicular long-term protocol group were significantly higher than those in the antagonist group [55.71% (122/219) vs. 37.82% (45/119), P=0.002; 63.58% (96/151) vs. 46.99% (39/83), P=0.014; 58.94% (89/151) vs. 39.76% (33/83), P=0.005]. In the overweight population, the implantation rate and the clinical pregnancy rate in early follicular long-term protocol group were significantly higher than those in the antagonist group [68.09% (32/47) vs. 43.40% (23/53), P=0.013; 81.25% (26/32) vs. 57.14% (20/35), P=0.034], but there was no statistically significant difference in the live birth rate ( P>0.05). Conclusion:Compared with the antagonist protocol, early follicular long-term protocol for unexplained infertility patients may achieve higher clinical pregnancy outcomes in IVF fresh cycle, but it could increased the duration of descending, the duration and dosage of Gn used.

Objective Using of cumulative live birth rate(CLBR)per oocytes retrieved cycle,to assess the clinical outcomes of in vitro fertilization or intracytoplasmic sperm injection(IVF/ICSI),and to explore impact factors on CLBR following utilization of all fresh and frozen embryos in one complete IVF/ICSI cycle using gonadotropin-releasing hormone(GnRH)agonist, GnRH-antagonist and clomiphene mild stimulation protocols. Methods Of the patients who underwent IVF/ICSI from January 1st, 2014 to December 31st, 2015 in the First Affiliated Hospital, Nanjing Medical University, a total of 6 142 oocytes retrieved cycles were included. The clinical and laboratory parameters of different ovarian stimulation protocols, and the effects of the age, number of oocytes retrieved and number of embryos available on the CLBR of each oocytes retrieved cycle were analyzed.Results The CLBR was 69.0%(2 004/2 906)in the GnRH-agonist protocol versus 67.4%(644/955)in the GnRH-antagonist protocol (P>0.05); the CLBR of clomiphene mild stimulation protocol was 53.2%(1 215/2 281),significantly lower than those of the other two protocols (all P<0.05). The CLBR significantly decreased with age increased. When divided into four groups according to the patients′ age, we found that CLBR were not statistically significant using three different protocols in the 20-25 years old group(all P>0.05).There was a strong association between the number of oocytes retrieved and embryos available on CLBR. CLBR rose significantly with an increasing number of oocytes up to 6, then the rising trend slowed down. Patients were categorized into four groups according to the number of oocytes retrieved,CLBR was significantly higher using GnRH-antagonist protocol (50.0%)than mild stimulation protocol(37.0%)in low ovarian responder(0-4 oocytes)group(P<0.05). The CLBR were no significant difference among three protocols in normal(10-15 oocytes)and high responders(≥15 oocytes)group(all P>0.05).The incidence rate of ovarian hyperstimulation syndrome in GnRH-agonist protocols(5.2%,152/2 906)were significantly higher than those of GnRH-antagonist(4.4%, 42/955)and clomiphene mild stimulation protocols(1.5%,34/2 281;all P<0.05).Conclusions CLBR is an important index to assess the clinical outcomes of IVF/ICSI. Age, number of oocytes retrieved and embryos available could affect CLBR obviously. According to the different age and ovarian response of patients, we should design ovarian stimulation protocols based on target oocytes number in order to get higher CLBR and reduce complications.

Objective To investigate the impact of dyslipidemia on in-vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) pregnancy outcome in patients with polycystic ovary syndrome (PCOS).Methods From July 2013 to March 2016,468 PCOS patients with antagonist protocol in IVF/ICSI of First Affiliated Hospital of Nanjing Medical University,cycles were divided into dyslipidemia group (108 cases) and normol blood lipids group (360 cases) according to the serum cholesterol,triglyceride (TG),high-density lipoprotein,low density lipoprotein levels.The general condition and clinical outcomes of the two groups were analyzed retrospectively,including the implantation rate,clinical pregnancy rate,live birth rate and the incidence of moderate to severe ovarian hyperstimulation syndrome (OHSS),etc.Besides,stratified analysis and multivariate logistic regression analysis were used to correct the impact of body mass index (BMI).Results (1) Comparing the based data of dyslipidemia group and normal blood lipids group:age,years of infertility,basic FSH,basic LH,basic estradiol and other indexes had no significant differences (all P＞0.05),but BMI of dyslipidemia group was significantly higher than normal blood lipids group [(25.0±3.0) versus (23.1±3.0) kg/m2],difference had statistical significance (P＜0.01).(2) The high score embryo rate,endometrial thickness on the day of hCG injection,progesterone and LH levels on the day of hCG injection,moderate to severe OHSS rate and miscarriage rate in the two groups did not exhibit remarkable differences (all P＞0.05).However,the number of dominant follicle,retrieved oocyte number,estrogen level on the day of hCG injection,implantation rate,biochemical pregnancy rate,clinical pregnancy rate and the live birth rate in dyslipidemia group were significantly less than those of normal blood lipids group (all P＜0.05),the dose of gonadotropin (Gn) and days of stimulation were significantly higher compared with the normal blood lipids group,there were significant differences statistically (all P＜0.05).(3) Stratified analysis showed that no matter in BMI＜24 or BMI≥24 kg/m2 group,the dose of Gn and days of stimulation were significantly higher in the dyslipidemia group than those of the normal blood lipids group,the difference was statistically significant (P＜0.05).However,the number of oocytes retrieved,estrogen level on the day of hCG injection had obvious downtrend,and the difference was statistically significant (P＜0.05) in BMI≥24 kg/m2 group.Multivariate logistic regression analysis found that,even after the correction of BMI,dyslipidemia still had negative impact on implantation rate,biochemical pregnancy rate,clinical pregnancy rate and the live birth rate (P＜0.05).(4) Further analysis of the different components of blood lipids in the clinical pregnancy group and unobtained pregnancy group revealed that the level of triglyceride (TG) in the unobtained pregnancy group was significantly higher than that in the pregnancy group,and the difference was statistically significant (P＜0.05);logistic regression analysis also showed that the increase of TG levels was negatively correlated with the clinical pregnancy rate of PCOS patients (P＜0.05).Conclusions PCOS patients combined with dyslipidemia have a higher BMI,and dyslipidemia increases the dosage of Gn,reduces the implantation rate,clinical pregnancy rate and live birth rate,especially the increase of TG level,which has adverse effects on IVF/ICSI outcome in patients with PCOS.

ABSTRACT:Objective To assess the effect of prolonged laparoscopic surgery on peritoneal mesothelial cells and fibrinolysis in humans.Methods We examined prospectively 1 6 consecutive patients who underwent laparoscopic surgery (LAP)and 2 1 patients who underwent conventional open surgery (OP)for high-medium rectal cancer with curative intent.During the procedure,biopsy of the parietal peritoneum was made before operation and at 45 min,90 min,and 120 min after operation.The tissue-type plasminogen activator (tPA)and plasminogen activator inhibitor-1 (PAI-1 )were determined by enzyme-linked immunosorbent assay in peritoneal tissues.The cellular injury was detected by LDH assay.The proliferation was quantified by MTT assay.Results PAI-1 activity in the peritoneal tissue was significantly lower in LAP group than in the OP group.tPA activity decreased after 45min of open surgery,but there was no significant change in the LAP group.With time extension,the LDH activity increased and the proliferation of the mesothelial cells decreased.Conclusion Preservation of a prolonged hypofibrinolytic state by inhibition of PAI-1 up-regulation during LAP may predispose patients to less postoperative peritoneal adhesion. The cellular injury becomes apparent and the proliferation is inhibited during prolonged laparoscopic surgery.