ObjectiveTo investigate the roles of 11 β-hydroxysteroid dehydrogenase type 1 ( 11 β-HSD1 )on hippocampus of rat with chronic unpredictable mild stress (CUMS).MethodsTwenty-four male SpragueDawley rats were randomly divided into control group and depressive model group. Chronic unpredictable mild stress (CUMS) was used to make up depressive animal model.Behavioral changes were recorded by body weight measuring,sucrose consumption test (SCT) and open field test (OFT),respectively.The mRNA transcription of 11β-HSD1 in hippocampus tissues of the rats were detected by real-time RT-PCR,and the protein expression of 11β-HSD1 were detected by western blot and immunofluorescence.ResultsBcforc starting CUMS protocol,the rats exhibited equivalent weight and sucrose consumption.Twenty-eight days after CUMS protocol,behavior parameters such as body weight,sucrose consumption,nunber of crossing,and number of rearing were significantly decreased in rats exposed to CUMS group compared with control group (P ＜ 0.05,P ＜ 0.01 ).Correspondingly,realtime RT-PCR assays showed the mRNA expression of 11 β-HSD1 in the hippocampus of CUMS group,which was (31 ±9) ％ lower than that of control group.Meanwhile,the protein expression of it in CUMS group was lower than that of control group (P ＜ 0.05 ).Inmunofluorescence revealed that the number of positive 11 3-HSD1 cells was high (223 ± 13) in the control group,while the number was decreased prominently (92 ± 11 ) in the CUMS group (P ＜ 0.01 ).ConclusionDepressive behavior of rats is induced and the expression of 11 β-HSD1 in the hippocampus is decreased prominently by CUMS,the mechanism of which is at least related to the low expression of 11β-HSD1 and disturbance of glucocorticoid metabolism caused by CUMS.

Objective To study the effect of brain-derived neurotrophic factor (BDNF) on the environmental nutrition and neural differentiation of the transplanted stem cells under hypothermia.Methods The BDNF gene mediated by liposome was transfected into 293T cell line, and ELISA assay was applied to find the peak time of BDNF expression. When BDNF was highly expressed, the supernatant was collected for establishment of SD rat models of brain injury. The rats were divided into Group A (stem cell transplantation group) and Group B (stem cell transplantation and BDNF group). Rats in both groups were under hypothermia treatment for five days. Four and eight days later ( three days from rewarming), rat brain tissues were obtained to detect the expressions of proliferating cell nuclear antigen (PCNA), nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) by immunohistochemical method and to detect the apoptosis by in situ hybridization. Finally, the nerve function scores were obtained for evaluation of the nerve function. Results The ELISA showed that the high level of BDNF expression was at 48 to 60 hours after gene transfection. PCNA and nestin were highly expressed, while NES and GFAP showed nil or low level of expression in both groups at the fourth day after hypothermia, with little apoptotic cells especially in the Group B (P ＜0.05). The expressions of PCNA and nestin were decreased, but the expressions of NSE and GFAP were increased at the third day after rewarming. The positive rate of NSE expression in the Group B was much higher and the apoptotic cells were much less compared with the Group A ( P ＜ 0. 05 ). A better nerve score was obtained in the Group B. Conclusion BDNF can enhance the survival rate of the transplanted stem cells and induce their differentiation into neurons under hypothermia.

ObjectiveTo investigate whether CD4+ CD25+ CD127dim/- regulatory T lymphocytes (Treg) can induce the proliferation of hepatic stellate cells (HSC) and expression of fibrosis-related factors on HSC in vitro and further to explore the mechanism of Treg inducing fibrogenesis. MethodsHSC LX-2 cells were subcultured.CD4+ CD25+ CD127dim/- cells were purified using magnetic cell separation. The HSC were co-cultured with Treg by direct contact or by Transwell system in vitro. The HSC cultured alone was used as control. Cell proliferation was measured by CCK-8 assay.The expression of transforming growth factor (TGF)-β1 was detected by enzyme-linked inmunosorbent assay (ELISA), and the expressions of hyaluronic acid (HA) and precollagen Ⅲ (PC Ⅲ ) were detected by radio immunoassay (RIA). The data were analyzed by LSD-t test. ResultsHSC proliferation was strongest when Treg∶ HSC= 1.5∶ 1. The absorbance in direct contact co-culture group and Transwell system co-culture group was (0. 713±0. 032) cpm and (0. 735±0. 028) cpm, respectively, both of which were higher than that in control group [(0. 677 ± 0. 029) cpm](t = 5. 4003 and 8. 7878,respectively; both P＜0. 01). The concentrations of TGF-β1 in the supernatant were (781. 59 ±76.45) pg/mL and (813. 53±60. 62) pg/mL, respectively in direct contact co-culture group and Transwell system co-culture group, which were significantly higher than that in control group [(722.51±59. 66) pg/mL](t = 4.0014 and 6. 1653, respectively; both P＜0.01).The concentrations of HA were (433. 575±27.90) ng/mL and (445.40±23.73) ng/mL, respectively in direct contact co-culture group and Transwell system co-culture group, which were higher compared to that in control group [-(415. 83±19.44) ng/mL](t =3. 3124 and 5. 5231, respectively; both P＜0.01). Likewise, the concentrations of PCⅢ were (21. 93± 1.71) and (23. 125± 1.87) ng/mL in direct contact group and Transwell group, respectively compared to (20. 10± 1.49) ng/mL in control group (t = 4. 8082 and 7. 9436, respectively; both P ＜ 0.01).Furthermore, the absorbance,concentrations of TGF-β1, HA and PC Ⅲ in Transwell co-culture group were all higher compared to direct contact group (t = 3. 3875, 2.1639, 2. 2107 and 3.1354, respectively; all P＜0. 05).ConclusionsThe cell proliferation and the expressions of fibrosis-related factors in HSC increase greatly after co-cultured with CD4+ CD25+ CD127dim/-Treg. Therefore, Treg may play an important role in inducing liver fibrogenesis.

Objective To investigate the dynamic expressions of exchange protein directly activated by cyclic adenosine monophosphate (cAMP) (Epac) in rat model of hepatic fibrosis(HF).Methods Forty-two male SD rats were divided into control group (n = 6) and model group (n = 36)which was divided into six subgroups of day 4, week 1, week 2, week 4,week 6 and week 8 with six rats in each subgroup. The rat model of HF was established by intraperitoneal injection of dimethylnitrosamine (DMN). The pathological changes of liver were observed by Hematoxylin-Eosin and Masson staining. Reverse transcription-polymerase chain reaction (RT-PCR),immunohistochemistry and Western blot were employed to detect the mRNA and protein expressions of Epac1, Epac2 and transforming gronth factor (TGF)β1 during the process of modeling and localization in the liver. The statistical analysis was done using one-factor ANOVA, LSD-t test,Dunnett T3 test and Pearson linear correlation analysis. Results Rat model of liver fibrosis was established successfully. In control group, Epac1 (0. 031 28±0. 008 96) and Epac2 protein (0.034 43±0. 002 45) mainly expressed in the cytoplasm of hepatocytes. In model group, the level of Epac1 decreased at day 4 (0. 023 97±0. 003 81) and week 1 (0. 015 81±0. 002 48) ,then began to increase at week 2 of modeling and peaked at week 6 (0. 039 54±0. 001 43), which had statistical significance compared to the control group (t= 5.47,11.58 and - 6.18, respectively; all P＜0.05). Epac2 protein expression declined after modeling, reached the lowest level at week 4 (0. 011 21 ±0. 001 32), which had statistical significance compared to the control group (t= 24. 50, P＜0. 05). TGFβ1 protein expression increased after modeling and peaked at week 4 (0. 011 30±0.001 03) which had statistical significance (t= -23. 36, P＜0. 05) compared to the control group (0. 002 08 ±0. 000 18). The expressions of Epac1, Epac2 and TGFβ1 mRNA were consistent with the trend of protein levels.Correlation analysis showed that Epac1 protein was positively correlated with the course of HF (r =0. 703, P＜0.01 ), while Epac2 protein was negatively correlated (r = - 0. 409, P＜0.05). Conclusions During the progression of HF, Epac1 expression tends to decrease firstly and increase afterwards,while Epac2 expression declines continually. Epac may be involved in the pathogenesis of HF.

Objective To investigate the roles of five scoring systems including model for endstage liver disease (MELD), Child-Turcotte-Pugh (CTP), Mayo, MESO and MELD-Na scoring systems, in predicting the prognosis of patients with chronic severe hepatitis. Methods The clinical data of 213 patients with chronic severe hepatitis were retrospectively studied. The five scoring systems were applied respectively to evaluate the scores in survival group and death group. The capability of these five scoring systems to predict the prognosis of severe hepatitis were compared by the receiver operating characteristic (ROC) curve, area under curve (AUC) and cut-off value.Measurement data were compared by group t test. The comparisons of AUC among scoring systems were done using MEDCLAC software. Results The scores of death group evaluated by MELD, CTP,Mayo, MESO or MELD-Na scoring systems (30.6 ± 9.5, 11.3 ± 1.5, 10.4 ± 1.3, 2.3 ± 0.8 and 39.0 ± 11.8, respectively) were consistently higher than those of survival group (21.1± 6.8, 10.6 ±1.6, 9.0±1.5, 1.6±0.5 and 22.6±8.2, respectively) (P＜0.01). The values of AUC of these five systems were 0.810, 0.623, 0.749, 0.829 and 0.885, respectively. The Youden's indexes of these five systems were 0.507, 0.175, 0.389, 0.528 and 0.650, respectively. Conclusions The CTP scoring systems can not predict the prognosis of chronic severe hepatitis very well. The Mayo scoring systems can partially predict the prognosis. On the contrary, MELD, MESO and MELD-Na systems can successfully predict the disease prognosis, and the score of MELD-Na system shows the best correlation with the prognosis.

Objective　To evaluate the therapeutic efficiency of reversed lamellar keratoplasty for bullous keratopathy, Terrien's marginal degeneration and ocular chemical burns.Methods　Twenty eyes of patients with bullous keratopathy, twelve eyes with Terrien's marginal degeneration, ten eyes with chemical burns were recruited in this study.Diseased tissues were removed by means of dissection.Reversed lamellae corneal tissues which had been prepared by cryopreservation were thawed and rinsed. Then the prepared posterior corneal lamellae was turned over, endothelium facing upwards. The diameter of the graft should be 0.5～1.0mm larger than that of the recipient bed. The reversed donor posterior lamellae was sutured into the host keratectomy bed with 10-0 nylon sutures continuously or interruptedly.Results　Allograft reaction episode occurred only one (chemical burn) of the total 42 eyes enrolled.The grafts of the rest 41 eyes maintained clear or semitransparent for 1～5 years. Graft edema could be observed within 2～4 weeks postoperatively. Postoperative interface separation was not found. For all 20 patients with bullous keratopathy, the symptoms of pain disappeared. The visual acuity improved 2～3 lines of Snellen among eleven of twelve patients with Terrien's marginal degeneration. The visual acuity improved 2～3 lines of Snellen with four patients with chemical burns.Conclusion　The technique of reversed lamellar keratoplasty can be applied successfully for tectonic reconstruction of corneal surface such as bullous keratopathy, Terrien' s marginal degeneration and also chemical burns.The effects of therapy was satisfactory.

Objective To investigate the changes and clinical significance of the frequency of circulating CD4+ CD25+ regulatory T cells (Treg) in nonalcoholic fatty liver (NAFL) patients.Methods CD4+ CD25+ Treg in the peripheral blood from 50 NAFL patients and 50 healthy subjects were quantitatively analyzed using flow cytometry. Group t test or Mann-Whitney U test and Spearman's rank correlation test were used for statistical analysis.Results The proportion of circulating CD4+CD25+ Treg in NAFL patients was (5.39 ± 1.94)％,which was significantly higher than that in healthy controls [(4.21±1.52)％](t=3.385,P＜0.01).Further analysis revealed that the frequency of Treg was positively correlated with triglyceride (TG) level and body mass index (r=0.307 and 0.251,respectively; P=0.002 and 0.012,respectively),and negatively correlated with high density lipoproteincholesterol (HDL-C) (r=-0.306,P=0.002).Meanwhile,Treg in patients with high body mass index,high TG,low HDL-C,hypertension and metabolic syndrome (MS) were all higher than those in controls (t=2.294,2.533,3.154,2.010 and 4.454,respectively; all P＜0.05).But there was no significant difference between patients with high fasting blood glucose and controls (U=1143.500,P=0.471).Conclusion The increased frequency of peripheral Treg in NAFL patients may have some relations with the imbalance of proinflammation and anti-inflammation in NAFL patients who coexisting with MS.

Objective:To investigate the clinical characteristics and possible mechanisms of remote intracranial hematoma (RIH) in patients with intracranial aneurysm after interventional embolization.Methods:Six patients with RIH from a series of 58 consecutive patients with intracranial aneurysm, admitted to and performed interventional embolization in our hospital from January 2016 and December 2018, were chosen in our study. Their clinical data were analyzed retrospectively and compared with those without RIH at the same period.Results:In these 6 patients, 4 had history of hypertension, 5 had aneurysm located in the internal carotid artery, 5 were treated with stents combined with postoperative routine anticoagulation treatment. The remote intracranial hematoma occurred within 7 d of interventional embolization, and the hematoma was located in the cerebral hemisphere on the same side of the aneurysm; 4 patients underwent intracranial hematoma puncture catheter drainage; 1 patient was treated conservatively, and one was treated by craniotomy. After treatment, 1 patient recovered (modified Rankin scale [mRS] score of 1), 1 patient had poor prognosis (mRS scores of 5) and discharged automatically, and the rest 4 patients (mRS scores of 3-5) left some degrees of neurological dysfunction. As compared with 52 patients without RIH, 6 patients with RIH had significantly higher percentages of patients used stents and postoperatively used anticoagulation, and higher percentages of patients with poor clinical outcomes at discharge ( P<0.05). Conclusion:Stent-assisted coil embolization in patients with internal carotid artery aneurysm combined with hypertension should be highly vigilant about the possibility of RIH.

Objective:To explore the accuracy of intelligent calculation (IC) method for risk assessment of hospitalization for patients, aiming to build a more advantageous risk assessment system.Methods:The "Search Engine" program was developed based on hospital information system (HIS) of the Fifth Center Hospital in Tianjin, which automatically captured patient information and generated nutritional risk screening 2002 (NRS 2002) score, Caprini thrombosis risk assessment model and Padua thrombosis risk assessment model for venous thromboembolism (VTE), the CHA 2DS 2-VASc for predicting stroke risk stratification in atrial fibrillation and the HAS-BLED for predicting bleeding risk in anticoagulated patients with atrial fibrillation. A randomized controlled trial was conducted. According to the applicable conditions of each risk assessment, 100 risk scores from "Search Engine" program belonged to each risk assessment were randomly selected, defined as the IC group. Manual scoring with the data of the same case at the same time, defined as the traditional calculation (TC) group, compared the consistency of the scores and the difference in time-consuming between the two groups. Results:The Bland-Altman plots showed that the 95% limits of agreement (95% LoA) of NRS 2002 score, Caprini score, Padua score, CHA 2DS 2-VASc score and HAS-BLED score was -0.46 to 0.41, -0.49 to 0.52, -0.50 to 0.41, -0.67 to 0.60, -0.44 to 0.43, respectively, all P > 0.05. In this study, the Bland-Altman plot showed that 95%, 96%, 97%, 97%, 95% plots fell within the 95% LoA in NRS 2002 score, Caprini score, Padua score, wwCHA 2DS 2-VASc score and HAS-BLED score by the two methods, respectively. The all plots of 95% LoA were within the clinically acceptable range (-0.5 to 0.5 scores). The time-consuming of NRS 2002 score, Caprini score, Padua score, CHA 2DS 2-VASc score and HAS-BLED score in IC group were significantly shorter than those in TC group [0.72 (0.71, 0.73) seconds vs. 361.02 (322.41, 361.02) seconds, 0.72 (0.72, 0.73) seconds vs. 196.68 (179.99, 291.20) seconds, 0.72 (0.72, 0.73) seconds vs. 105.75 (92.32, 114.70) seconds, 0.72 (0.71, 0.72) seconds vs. 72.66 (56.24, 84.20) seconds, 0.72 (0.71, 0.72) seconds vs. 51.30 (38.88, 57.15) seconds, respectively, all P < 0.001]. Conclusion:For the above five risk assessments, the TC method and IC method has good consistency in scores, and the IC method is faster, which has good application prospect for clinical application.

Objective:To explore the characteristics of the changes in risk score for intensive care unit (ICU) patients during hospitalization by the intelligent calculation method, and to provide evidence for the risk prevention.Methods:In this retrospective study, ICU patients of the Fifth Central Hospital in Tianjin from November 3, 2021 to March 28, 2022 were enrolled and divided into ≥ 14 days group, 10-13 days group, 7-9 days group, and 3-6 days group according to the ICU length of stay. Risk scores assessed by the intelligent calculation method of the ICU patients were collected, including nutritional risk screening 2002 (NRS 2002), Caprini score and Padua score. NRS 2002 score for all patients, Caprini score for surgical patients and Padua score for internal medicine patients were selected. Trends in change of each score were compared between patients admitted to ICU 1, 3, 7 (if necessary), 10 (if necessary), and 14 days (if necessary).Results:A total of 138 patients were involved, including 79 males and 59 females, with an average age of (61.71±18.86) years and an average hospital stay of [6.00 (4.00, 9.25)] days. ① in the group with ICU length of stay ≥ 14 days (21 cases): there was no significant change in the NRS 2002 scores of the patients within 10 days, but the NRS 2002 score was significantly decreased in 14 days as compared with 1 day [3.00 (2.50, 3.50) vs. 4.00 (3.00, 5.00), P < 0.05]; both Caprini and Padua score were increased with prolonged hospital stay and compared with 1 day, the scores at the other time points were significantly increased, especially at 14 days [Caprini score: 5.00 (3.25, 7.00) vs. 2.50 (1.25, 5.50), Padua score: 6.00 (6.00, 7.00) vs. 3.00 (1.00, 3.00), both P < 0.05].② in the group with ICU length of stay from 10-13 days (15 cases): with the prolonged hospital stay, there was no significant change in NRS 2002 score, but both Caprini and Padua score were increased at 3, 7, 10 days, especially at 10 days [Caprini score: 3.00 (2.00, 4.75) vs. 2.00 (0.25, 2.75), Padua score: 5.00 (3.50, 6.00) vs. 2.00 (0.50, 4.00), both P < 0.05].③ in the group with ICU length of stay from 7-9 days (23 cases): compared with 1 day, the NRS 2002 score at 3 days and7 days were decreased, but the Caprini and Padua score were increased, especially at 7 days [NRS 2002 score: 2.00 (1.00, 4.00) vs. 2.00 (2.00, 4.00), Caprini score: 3.00 (2.00, 5.50) vs. 2.00 (0.25, 3.00), Padua score: 5.00 (4.00, 6.00) vs. 2.00 (0, 2.00), all P < 0.05]. ④ in the group with ICU length of stay from 3-6 days (79 cases): compared with 1 day, the NRS 2002 score at 3 days was decreased [NRS 2002 score: 2.00 (1.00, 3.00) vs. 2.00 (1.00, 3.00), P < 0.05], Caprini and Padua score were significantly increased [Caprini score: 3.00 (2.00, 4.00) vs. 2.00 (1.00, 3.00), Padua score: 5.00 (4.00, 5.00) vs. 2.00 (1.00, 3.00), both P < 0.05]. Conclusion:Based on dynamic assessment of intelligent calculation methods, the risk of thrombosis in ICU patients increased with hospital length of stay, and the nutritional risk was generally flat or reducing in different hospitalization periods.