BACKGROUND: Vitrification is a comparatively new technology which applies high concentration cryoprotectant and rapid refrigeration. By the method, the cells were quickly frozen and to avoid damage by ice crystals inside and outside. OBJECTIVE: To compare the effect of four cryoprotectants on morphology and function of ovarian tissue in rats after vitrification. METHODS: The rats were randomly assigned into six groups with 6 rats for each: DMSO + EG, DMSO + EG + sucrose, DMSC +EG + sucrose + acetamide, EG + sucrose + acetamide, ovariectomized, and normal control groups. The ovarian tissues of four freezing groups were treated with the corresponding cryoprotectants, the vitrified ovarian tissues were then resected but not frozen and transplanted; otherwise, tissues were not treated with any treatment in the normal control group. Two weeks after freezing, the tissues were thawed and heterotopic-transplanted into femoribus intemus of hind limb. At 30 days after implantation, vaginal epithelial cells and estrus cycle were observed, while after three months, blood were collected to detect the level of estradiol (E2) and the ovarian tissues were reclaimed to analyze their morphological changes. RESULTS AND CONCLUSION: All ovarian tissues were damaged after cryoprersarvation in four freezing groups. The rates ot healthy primordial follicles were 67.9%, 71.6%, 80.5%, and 59.4%, respectively, while healthy primary follicles were 41.6%, 52.3%, 55.9%, and 36.7%, respectively. In all freezing groups, the rate of the healthy follicles in DMSO + EG + sucrose + acetamide group was higher than DMSO + EG group and EG + sucrose + acetamide group (P < 0.05). No significant difference was found in the proportion of follicles at different development stages among four groups. The typical secondary follicle was not found in four groups. Damaged ovotid showed oocyte pyknosis and vacuolation in cytoplasmic area. There was not typical cell type of all freezing groups. Ovarian autografting gained visible vascularity from surrounding tissue that connected ovarian tissue to form net. There was a lot of blood capillary in transplanted ovarian tissues and clumped primordial follicles in cortical substance. The rates of primary follicles and secondary follicles were lower than primordial follicles. The level of serum estradiol was obviously decreased compared with normal control group (P < 0.01). There was significant difference between DMSO + EG + sucrose + acetamide group and other three freezing groups (P < 0.05). Four kinds of freezing methods have poor effects on different stages of follicles and the structure of ovariarn tissue. DMSO + EG + sucrose + acetamide group is an optimal protocol for cryoprerserving ovarian tissue. Freezing methods still need to explore further because the rats had not appeared disciplinary estrus cycle after ovarian autoqrafting.

The experiment was performed at the Institute of Treatment and Prevention of Tumor of Jilin Province and Department of Etiology, Jilin Medical College from October 2004 to June 2005. The normal umbilical cord blood (UCB) was provided by a healthy lying-in woman from Changchun Hospital with the permission of the pregnant woman. Full-term normal delivery and UCB of healthy pregnant women was collected by aseptic venepuncture after fetal disengagement, and with Natrium Citricum anticoagulation. After the UCB mononuclear cells were isolated by Ficoll-paque (relative volume mass: 1.077?0.22) density gradient centrifugation, long-term liquid culture system was used for UCB adherent cells, and co-cultured with UCB mononuclear cells. The adherent cells were observed and mononuclear cell cycles were tested with flow cytometer. It showed that adherent layer of UCB increased UCB mononuclear cells expansion as compared with that of non-adherent layer of UCB after co-culture for 7 days. The cell percentage in S+G2+M phase obviously increased, and there was significant difference [(42.7?1.1)%,(35.5?2.8)%,P

AIM: To study the effect of the radix sophorae flavescentis on cellular immunity in rats with Pneumocystis Carinii Pneumonia (PCP) induced by long-term use of immunosuppressant, and explore the action of traditional Chinese medicine for the immunological regulation and infectious prevention after organ transplantation. METHODS: The experiment was conducted at Department of Pathobiology, Jilin Medical College from May 2005 to March 2006. Forty adult healthy female SD rats were selected from Harbin Medical University (Certification: 02473146) and randomly divided into experiment group and control group, with 20 rats in each. The model of PCP was set up by glucocorticoid injection subcutaneously to SD rats (25 mg once, 2 times/week). The mixture of sophorae flavescentis was given to stomach with tube in experiment group (3 mL/kg, 2 times/day), and was consisted of radix sophorae flavescentis, ash bark, amur cork-tree, malt, milkvetch root and danshen root. Six weeks later, all the rats were anesthetized and broncholveolar lavage fluids were collected.①Alveolar washing fluid was concentrated 10 times and the levels of the soluble interleukin-2 receptor (sIL-2R) were examined by double antigen sandwich ELISA.②Blood was sampled from rat eyes and the count of lymphocytes in peripheral blood were detected.③The percentages of CD3+ and CD4+ subgroups were assessed with erythrocyte chaplet kit sensitized by antigen. RESULTS: All 40 rats were involved in the result analysis without drop.①The count of lymphocytes in peripheral blood in experiment group was significantly higher than that in control group (5.1?1.3)%, (0.8?0.3)%, P

Objective: To investigate the effects of chemokine like factor 1 (CKLF1) on proliferation and metabolism of chondrocytes. Methods: Cell culture, 3H TdR and 3H Proline incorporation methods were used. Chondrocytes were harvested from rabbit articular cartilage. First passage cells were seeded on 96 well plates. After synchronization,the medium was replaced by DMEM containing 5% FCS with various concentration of CKLF1 conditioned medium. The synthesis of mucopolysaccharide was detected by Saffraan O staining. The transcription of inducible nitricoxide synthase (iNOS) was detected by semi quantitative RT-PCR. Results: CKLF1 inhibited the DNA, collagen and mucopolysaccharide synthesis significantly, meanwhile, stimulated the transcription of iNOS. Conclusion: CKLF1 inhibits the proliferation and matrix synthesis of chondrocytes, which might be an important factor resulting in cartilage destructive lesions. CKLF1 may exert its effects on chondrocytes through iNOS pathway.

Objective: To characterize the expression of programmed cell death 5 (PDCD5) in normal and osteoarthritic human cartilage. Methods: Articular cartilage specimens were obtained from 20 patients with osteoarthritis and 10 with femoral neck (normal cartilage) at the time of arthroplasty. Expression of PDCD5 was detected by flow cytometry, immunofluorescence, RT-PCR and immunohistochemical analysis. Results: PDCD5 expression in osteoarthritis cartilage was significantly higher than in normal cartilage especially in the nucleus. PDCD5 positive chondrocytes were mainly observed in the superficial and deep zone of osteoarthritis tissue sections,and in contrast, in the superficial and middle regions of normal controls. Conclusion: Since apoptotic chondrocyte death occurs more frequently in osteoarthritis compared to normal cartilage and PDCD5 is an apoptosis related protein, the different expression patterns of PDCD5 in osteoarthritis and normal cartilage suggest that it might be involved in the pathogenesis of osteoarthritis.

OBJECTIVE:To investigate the effects of simvastatin intensive treatment on the Postoperative Related Indexes of patients with acute coronary syndrome(ACS) underwent percutaneous coronary intervention(PCI). METHODS:106 patients with were included in the study and randomly divided into observation group(53 cases)and control group(53 cases). Both groups were given aspirin 100 mg,qd+clopidogrel 75 mg,qd before PCI for 4 weeks;observation group was additionally given Simvastatin tablet orally 20 mg before supper 15 d before surgery. TC,TG,LDL-C,HDL-C,hs-CRP,IL-6 and IL-18 levels,LVEF,the occurrence of coronary artery restenosis were detected in 2 groups before surgery and 6 months after surgery. The occurrence of ADR was recorded during treatment. RESULTS:There was no statistical significance in the levels of TG,TC,LDL-C and HDL-C between 2 groups before surgery and 6 months after surgery (P>0.05). There was no statistical significance in hs-CRP,IL-18, IL-6 and LVEF levels between 2 groups before surgery(P>0.05). 6 months after surgery,hs-CRP,IL-6,IL-18 and LVEF levels of 2 groups were significantly higher than before treatment;hs-CRP,IL-6 and IL-18 levels of observation group were significantly lower than those of control group,and LVEF was significantly higher than control group,with statistical significance (P<0.05). The incidence of coronary artery restenosis in observation group was significantly lower than control group, with statistical significance (P<0.05). No ADR was found in 2 groups during treatment. CONCLUSIONS:Preoperative simvastatin intensive treatment can effectively reduce cardiovascular inflammation degree in patients with ACS after PCI,prevent the formation of coronary artery thrombus,and reduce the incidence of coronary artery restenosis so as to effectively improve the prognosis and don' t increase the incidence of ADR.

Objective To investigate the changes of serum concentration of amylase(AMY) and pathological of pancreas in severe acute pancreatitis of SD-rats treated with Ulinastatin combined with qingyitang and to provide the basis for clinical application.Methods Ninty male SD-rats with average weight of(271.59±11.13) g were conventional feeding for a week.Twelve hours before experiment,they were forbidden to take food.They were randomly divided into five groups(with 18 in every group):sham operation group,severe acute pancreatitis group,Ulinastatin treated group,qingyitang treated group,Ulinastatin combined with qingyitang treated group.Then eighteen rats in every group were divided randomly into 6 h,12 h and 24 h groups with six rats in each group.Severe acute pancreatitis model was created by retrograde injection of 5％ sodium taurocholate (0.1 ml/100 g) through pancreatic duct,in sham operation group,only pancreas were flipped.The three treated groups were treated with vein injection of Ulinastatin(20 000 U/kg)or qingyitang perfusion(1 ml/100 g)or Ulinastatin combined with qingyitangat 0 h,5 h and 11 h after severe acute pancreatitis model successfully created.AMY and patholigical changes of pancreas were measured.Results There was significant difference on AMY level at 6 h,12 h and 24.h between severe acute pancreatitis group and sham operation group[6 h:(16 518±327)U/L vs(1671±175)U/L,F =3296.24,P ＜0.01;12 h:(16 796±338) U/L vs(1751±223)U/L,F =3802.44,P ＜0.01;24 h:(1554±373) U/L vs(1724±187) U/L,F =2382.54,P ＜0.01].Compared with severe acute pancreatitis group,the AMY level and pathologic score both decreased significantly in Ulinastatin treated group,qingyitang treated group and Ulinastatin combined with qingyitang treated group(P ＜0.05 or P ＜ 0.01).At the same time,the AMY level and pathologic score both decreased more in Ulinastatin combined with qingyitang treated group than Ulinastatin treated group and qingyitang treated group(P ＜0.05 or P ＜ 0.01).Conclusion Combination of Ulinastatin with qingyitang in treatment of severe acute pancreatitis of rats is superior to treatment with single medicine.

Objective:To evaluate the correlation between the timed up and go(TUG)test and fall risks in elderly frail patients.Methods:From July to September 2019, elderly frail patients who were treated at the cardiovascular department of our hospital were enrolled.Basic clinical data and fall-related information of patients were collected.Patients were divided into the fall group and the non-fall group.Results on the body mass index(BMI), TUG, 4-meter maximum walking speed(4 m MWS)and Barthel index were compared between the two groups.The correlation between TUG and each indicator was examined.Multivariate Logistic regression analysis was used to analyze the correlation between the TUG and falls in elderly patients.Results:A total of 96 eligible patients were enrolled, including 35 in the fall group and 61 in the non-fall group.The average TUG time was longer in the fall group than in the non-fall group(16.45±6.44 s vs.10.17±2.91 s, t=-6.556, P<0.001). The correlation analysis results showed that the TUG was correlated with falls and 4 m MWS( r=0.582 and 0.875, both P<0.001). Multivariate Logistic regression analysis showed that the TUG( OR=1.201, 95% CI: 1.111-1.470, P=0.004)and 4 m MWS( OR=1.146, 95% CI: 1.063-1.244, P=0.015)were risk factors for falls. Conclusions:The TUG is correlated with fall risks in elderly frail patients and should be recommended as a routine test in clinical practice.

Objective:To analyze the clinical features, prevention, and treatment of diabetic ketoacidosis (DKA) in pregnancy.Methods:Clinical features, treatment, and pregnancy outcomes of ten pregnant patients diagnosed with DKA and treated at Peking University First Hospital from January 2004 to December 2021 were analyzed retrospectively using descriptive statistical analysis.Results:DKA was found in two cases in the first trimester, three in the second trimester, and five in the third trimester, respectively. Six had type 1 diabetes (including two patients with fulminant type 1 diabetes) and four patients had type 2 diabetes. Only two cases received regular pregnancy examinations, and two began prenatal care in the second trimester. The other six cases neither had prenatal examination nor regular examination. Infection, pancreatitis, and preeclampsia occurred in six, two, and one case, respectively, and among them, four had urinary system infection. Laboratory examination showed that the average pH, blood glucose, and glycated hemoglobin A1c (HbA1c) at onset were 7.06±0.16, (23.7±2.6) mmol/L and (9.2±0.8)%. All patients had different degrees of electrolyte disorder and tested positive for urine ketone bodies. After rehydration, glucose-lowering, and correction of electrolyte disorder, the acidosis was relieved, and the ketone bodies turned negative in all patients. There was no maternal death. The fetal loss occurred in five cases, including one with intrauterine fetal death in late pregnancy, one with embryo arrest in early pregnancy, one with unavoidable abortion, and two terminated on maternal request. The remaining three were term labor and two were preterm labor.Conclusions:The leading cause of DKA in pregnancy is poor control of diabetic blood glucose, followed by infection. Once DKA is complicated during pregnancy, the outcome of the mother and her baby is poor. Its occurrence should be actively prevented. For women with fulminant type 1 diabetes, DKA is easier to occur, and the prognosis is poorer, so strict management and follow-up should be warranted.

Objective : To select and obtain vaginal Lactobacillus strains with inhibitory effect on Gardnerella vagi⁃ nalis , and to explore the possible mechanism of inhibition of Gardnerella vaginalis by Lactobacillus vaginalis strains , so as to provide a prerequisite for the development of dominant strains for human disease treatment. Methods : Gardnerella vaginalis and vaginal Lactobacillus strains were isolated from vaginal secretions of patients with bacterial vaginosis (BV) and healthy women , respectively. The Lactobacillus strains with the ability to inhibit the proliferation of Gardnerella vaginalis were screened by spot on lawn. Double agar diffusion method was used to compare the inhibitory effects of Lactobacillus solutions , cell⁃free culture supernatants (CFCs) and bacteria on Gardnerella vaginalis. Results : Sixty⁃two strains of vaginal Lactobacillus strains were isolated and purified from vaginal vaginal secretions of 10 patients with bacterial vaginosis. Among the 62 strains of vaginal Lactobacillus , 18 strains could inhibit the proliferation of Gardnerella vaginalis. The vaginal Lactobacillus solutions , the CFCs , and bacteria all had inhibitory effect on Gardnerella vaginalis. The inhibitory effects of vaginal Lactobacillus solutions were higher than those of the CFCs. After the CFCs were treated with proteinase K , the inhibitory effect of eight samples disappeared , and that of 10 samples weakened obviously. Conclusion In this paper, 62 vaginal Lactobacillus strains are purified and identified , of which 18 strains can inhibit the proliferation of Gardnerella vaginalis. The production of antimicrobial protein may be involved in the mechanism that vaginal Lactobacillus inhibits the proliferation of Gardnerella vaginalis.