Background and aim:Hepatitis B virus(HBV)-related gestational acute-on-chronic liver failure(ACLF)is a severe condition with limited treatment options.This study aimed to evaluate the efficacy and ideal timing of plasma exchange and continuous renal replacement therapy(CRRT)in managing pregnant women with HBV-related ACLF. Methods:This study retrospectively analyzed 51 eligible patients with HBV-related gestational ACLF between 2009 and 2020.Patients admitted to the study were divided into a conventional treatment group and a new treatment group according to whether they received the new management protocol,which included more aggressive plasma exchange(PE)and CRRT strategies.All 19 pregnant women with hepatic encephalopathy(HE)were divided into an early treatment group and a non-early treatment group according to whether PE therapy was initiated within three days.Our study had two primary objectives.Firstly,we aimed to evaluate the impact of PE and CRRT on puerperal survival.Secondly,we sought to assess the effects of early PE and CRRT regimens on puerperal survival in women with HE. Results:The levels of total bilirubin on the second day postpartum(D3),the third day postpartum(D4),and the fifth day postpartum(D6)were significantly lower in the new treatment group compared to the conventional treatment group(P=0.02,0.01,and 0.02,respectively).The ALT of D3 was significantly elevated in the new treatment group compared to the conventional treatment group(P=0.02).The incidence of HE overall increased from prenatal to postpartum D4,peaked on D4,and then gradually decreased from the fourth day postpartum(D5)(P=0.027).The first week after delivery revealed a significant difference in survival rate between the two groups,the conventional treatment group had statistically higher mortality rates compared to the new treatment group(P=0.002).Similarly,the entire puerperal period mortality rate of the conventional treatment group was statistically higher than the new treatment group(P=0.002).Moreover,among all patients with HE,the non-early treatment group showed significantly higher puerperal mortality rates compared to the early treatment group(P=0.006). Conclusions:Early PE and CRRT conducted within three days post-childbirth,enhance puerperal prog-nosis for HBV-related gestational ACLF.

Objective:To compare oxytocin combined with ergometrine with oxytocin alone in terms of primary prophylaxis for postpartum hemorrhage (PPH) at the time of cesarean section (CS).Methods:This was a multicenter double-blind randomized controlled interventional study comparing ergometrine combined with oxytocin and oxytocin alone administered at CS. From December 2018 to November 2019, a total of 298 parturients were enrolled in 16 hospitals nationwide. They were randomly divided into experimental group (ergometrine intra-myometrial injection following oxytocin intravenously; 148 cases) and control group (oxytocin intra-myometrial injection following oxytocin intravenously; 150 cases) according to 1∶1 random allocation. The following indexes were compared between the two groups: (1) main index: blood loss 2 hours (h) after delivery; (2) secondary indicators: postpartum blood loss at 6 h and 24 h, placental retention time, incidence of PPH, the proportion of additional use of uterine contraction drugs, hemostatic drugs or other hemostatic measures at 2 h and 24 h after delivery, the proportion requiring blood transfusion, and the proportion of prolonged hospital stay due to poor uterine involution; (3) safety indicators: nausea, vomiting, dizziness and other adverse reactions, and blood pressure at each time point of administration.Results:(1) The blood loss at 2 h after delivery in the experimental group [(402±18) ml] was less than that in the control group [(505±18) ml], and the difference was statistically significant ( P<0.05). (2) The blood loss at 6 h and 24 h after delivery in the experimental group were less than those in the control group, and the differences were statistically significant (all P<0.05). There were no significant differences between the two groups in the incidence of PPH, the proportion of additional use of uterine contraction drugs, hemostatic drugs or other hemostatic measures at 2 h and 24 h after delivery, the proportion requiring blood transfusion, and the proportion of prolonged hospital stay due to poor uterine involution (all P>0.05). (3) Adverse reactions occurred in 2 cases (1.4%, 2/148) in the experimental group and 1 case (0.7%, 1/150) in the control group. There was no significant difference between the two groups ( P>0.05). The systolic blood pressure within 2.0 h and diastolic blood pressure within 1.5 h of drug administration in the experimental group were higher than those in the control group, and the differences were statistically significant ( P<0.05), but the blood pressure of the two groups were in the normal range. Conclusion:The use of ergometrine injection in CS could reduce the amount of PPH, which is safe and feasible.

Objective:To observe and investigate the effect of HIF-2α in the process of neovascularization of proliferative diabetic retinopathy (PDR).Methods:Retrospective clinical study. From July 2014 to July 2015, 60 eyes of 57 PDR patients diagnosed in Ophthalmology Department of Affiliated Hospital of Qingdao University were included in the study. Twenty-eight eyes of 27 patients received intravitreal injections of 0.5 mg ranibizumab (0.05 ml) at 2-7 days before surgery (ranibizumab group) and other 32 eyes of 30 patients did not (group without ranibizumab). Eighteen eyes of 18 patients with epiretinal membranes were included as controls. Pathological specimens of PDR fibrovascular membrane and premacular membrane were obtained during vitrectomy. The immunohistochemical staining and real-time PCR (RT-PCR) were used to detecting the expression of HIF-2α, Dll4 and VEGF. Kruskal-wallis test was used to compare the expression differences of correlation factors between groups. Spearman correlation analysis was used to analyze the relationship between the two variables.Results:The immunohistochemical staining revealed that there were positive expression of HIF-2α, Dll4 and VEGF in all PDR membranes, regardless of the injection of the ranibizumab. The levels of HIF-2α, Dll4 and VEGF protein in the group without ranibizumab were higher than those of the ranibizumab group ( t=4.36, 6.01, 4.82; P=0.000, 0.008, 0.016). RT-PCR showed that the differences of the mRNA expression of HIF-2α, Dll4 and VEGF were all statistically significant among the PDR patients and controls ( H=18.81,19.60, 20.50; P=0.000, 0.000, 0.000). The expression of HIF-2α, Dll4 and VEGF in the PDR membranes was higher than that of epiretinal membranes from non-diabetic patients. In the PDR patients,the expression of HIF-2α, Dll4 and VEGF of the group without ranibizumab was higher than that of the ranibizumab group. The spearman correlation analysis showed that the expression of mRNA between HIF-2α and Dll4, HIF-2α and VEGF were both significantly correlated ( r=0.95, 0.87; P<0.05). Conclusions:The expression of HIF-2α in the PDR membranes was higher than that of the controls. It is positively correlated with the expression of the DLL4 and VEGF.

Objective To observe the expression ofprobucol on high glucose-induced specificity protein 1 (SP 1),kelchlike ECH associated protein 1 (Keap 1),NF-E2-related factor 2 (Nrf2) and glutamatecysteine ligase catalytic (GCLC) in the cultured human müller cells and preliminary study the antioxidation of the probucol on müller cells.Methods Primary cultured human müller cells were randomly divided into four groups:normoglycaemia group (5.5 mmol/L glucose),normoglycaemia with probucol group (5.5 mmol/L glucose+100 μmol/L probucol),hyperglycemia group (25.0 mmol/L glucose),hyperglycemia with probucol group (25.0 mmol/L glucose + 100 μmol/L probucol).Immunofluorescence staining was used to assess distribution of SP1,Keapl,Nrf2,GCLC in human Müller cells.SP1,Keapl,Nrf2 and GCLC messenger RNA (mRNA) expression was evaluated by quantitative real-time RT-PCR (qRT-PCR).Independent sample t test was used to compare the data between the two groups.Results All müller cells expressed glutamine synthetase (＞ 95％),which confirmed the cultured cells in vitro were the purification of generations of müller cells.The expressions of SP 1,Keap 1,Nrf2,and GCLC protein were positive in human müller cells.qRT-PCR indicated that SP1 (t=28.30,P＜0.000),Keap1 (t=5.369,P=0.006),and Nrf2 (t=10.59,P=0.001) mRNA in the hyperglycemia group increased obviously compared with the normoglycaemia group;GCLC (t=4.633,P=0.010)mRNA in the hyperglycemia group decreased significantly compared with the normoglycaemia group.However,SP1 (t=12.60,P=0.000) and Keapl (t=4.076,P=0.015) in the hyperglycemia with probucol group decreased significantly compared with the hyperglycemia group;Nrf2 (t=12.90,P=0.000) and GCLC (t=l 5.96,P＜0.000)mRNA in the hyperglycemia with probucol group increased obviously compared with with the hyperglycemia group.Conclusion Probucol plays an antioxidant role by inhibiting the expression of SP 1,Keap 1 and upregulating the expression of Nrf2,GCLC in müller cells induced by high glucose.

Objective To observe the influence of human umbilical cord mesenchymal stem cells (hUCMSC) transplanted into the tail vein of diabetic rats on apoptosis of retinal neurons and the retinal expression level of glial fibrillary acidic protein (GFAP).Methods Seventy clean male Sprague-Dawley rats were randomly divided into the normal control group (group A),diabetes mellitus (DM) only group (group B),DM + balanced salt solution (BSS) group (group C),DM + hUCMSC group (group D),with 10 rats in each group.DM rats were induced by intraperitoneal injection of streptozotocin.Apoptosis of retinal cells was assayed by dUTP nick end labeling.Immunohistochemistry and Western blot was performed to detect the retinal expressions of GFAP in rats.Results Compared with group A,large numbers of apoptotic cells could be found in the retinal ganglion cell layer (GCL) and inner nuclear layer (INL) of group B and group C,however the apoptotic cells in group D were significantly reduced than group B and C.The expression of GFAP was mainly located in the retinal GCL and retinal nerve fibre layer (RNFL) in group A,throughout the inner plexiform layer (IPL) in group B and C,only distributed in RNFL and GCL in group D.It was obvious that the expression of GFAP in group B and C was higher than group A.Compared with group B and C,the expression of GFAP in group D was significantly reduced.The difference of GFAP expression among the 4 groups was significant (F=79.635,P＜0.05).Conclusion hUCMSC could inhibit the apoptosis of retinal cells and activation of glial cells in early DM rats.

Objective To observe the effect of macular retinal thickness (CMT) on the long-term visual prognosis after intravitreal injection of Conbercept combined with retinal laser photocoagulation for macular edema (ME) secondary to branch retinal vein occlusion (BRVO).Methods A retrospective non randomized controlled study.Forty-one patients (41 eyes) of ischemic BRVO secondary ME were included in the study.Among them,there were 23 males (23 eyes) and 18 females (18 eyes).The average age was (56.49±8.94) years.The best corrected visual acuity (BCVA) and optical coherence tomography were performed.The mean logMAR BCVA was 0.82±0.41,and the mean CMT was (512.61 ± 185.32) μrn.According to the CMT reduction value at 1 month after treatment,the eyes were divided into no response group and response group,each has 15 patients of 15 eyes and 26 patients of 26 eyes respectively.The age and sex composition of the two groups were not statistically significant (t=-0.298,-1.708;P=0.767,0.096),and the difference of frequency of injection was statistically significant (t=3.589,P=0.010),and there was no statistical difference between the patients with logMAR BCVA and CMT (t=2.056,-1.876;P=0.460,0.070).The average follow-up was 8 months.The logMAR BCVA on 6 months after treatment was defined as long term vision.The changes of long term vision and CMT on 1 and 6 months of two groups after treatment were observed.Pearson correlation analysis showed that the correlation between long-term vision and age,logMAR BCVA before treatment,CMT before treatment,frequency of injection,and CMT value decreased 1 month after treatment.The correlation of long-term visual acuity with age,sex,logMAR BCVA before treatment,CMT before treatment,number of drugs before treatment,CMT reduction at 1 month after treatment,integrity of ellipsoid band and integrity of external membrane (ELM) were analyzed by multiple regression analysis.Results On 1 month after treatment,the CMT of the eyes was lower than that before treatment (231.48± 177.99) μm,and the average integrity of ELM and ellipsoid were 0.56±0.50 and 0.41 ±0.50 respectively.On 6 months after treatment,the average logMAR BVCA of the eyes was 0.48 ± 0.34.The results of Pearson correlation analysis showed that the long-term vision was positively correlated with the logMAR BCVA before treatment and the number of CMT reduction and the number of drug injection at 1 month after treatment (P＜0.05);there was no correlation with age and CMT before treatment (P＞0.05).The results of multiple regression analysis showed that the long-term vision was associated with logMAR BVCA before treatment,CMT reduction,ELM integrity,and the number of times of injection (P＜0.05),and no correlation with age,sex,CMT before treatment and the integrity of the ellipsoid (P＞0.05).On the 6 months after treatment,the logMAR BCVA in the non-response group and the response group were 0.86±0.23 and 0.26±0.14,and the average CMT was respectively (398.93±104.87) and (255.15 ± 55.18) μrn,and the average injection times were respectively (2.53 ± 1.46) and (1.31 ± 0.74) times.The average logMAR BCVA,CMT and injection times of the two groups were statistically significant (t=10.293,5.773,3.589;P=0.000,0.000,0.001).No complications related to drug or intravitreal injection occurred in all patients.Conclusion The long-term vision of ME secondary to BRVO after intravitreal injection of Conbercept combined with retinal laser photocoagulation was associated with the decrease of CMT and the integrity of the ELM after 1 month of treatment,no correlation was found between CMT and ellipsoid integrity before treatment.

Objective To observe the effect of Delta-like ligand 4 (Dll-4) on the pathological structure of retina in early diabetic rats (DM) and its relationship with vascular endothelial growth receptor-2 (VEGFR-2).Methods A total of 70 male Sprague-Dawley rats were randomly divided into normal group and DM group,with 10 and 60 rats in each group,respectively.The rats of DM group was induced by intraperitoneal injection of streptozotocin to established DM model.The rats with blood glucose recovery and death were excluded,and the final 60 rats were included in the statistics.Rats in the normal group were injected with an equal volume of citric acid-sodium citrate buffer.Rats in the DM group were divided into DM 1 month (DM lm) group,DM 2 months (DM 2m) group,DM 3 months (DM 3m) group and DM 3m + Anti group,DM 3m + phosphate buffer solution (PBS) group by random number table method,and 10 rats in each group.In the DM 3m+Anti group,4 μl ofantiDll-4 polyclonal antibody was injected into the vitreous cavity,and the antibody concentration was 0.25 mg/ml.The DM 3m+PBS group was intravitreally injected with an equal volume of PBS.Five days after the injection,the rats were sacrificed.Rats in the DM 3m group and the normal group were not treated,and were sacrificed 3 months after the model was established.The structure and microvascular changes of the retina were observed by hematoxylin-eosin staining,and the total thickness of the retina was measured.The expression of Dll-4 and VEGFR-2 in the retina was detected by immunohistochemistry and fluorescence quantitative polymerase chain reaction (PCR).One-way analysis of variance was used to compare the expression of Dll-4 and VEGFR-2 in the retina of each group.The least significant difference t test was used to compare the two groups.Results Light microscopy showed that the retinal ganglion cells layer in the DM 3m group were obviously edematous,the inner and outer nuclear layers were thinner,the number of cells was reduced,the arrangement was disordered,the edema of outer plexiform layer was obvious,and the microvessels were abnormally dilated.In the DM 3m+Anti group,the edema of outer plexiform layer was lessened than that of the DM 3m group,and the other layers were not significantly different from the DM 3m group.Compared with the normal group,the total retinal thickness of the DM 3m group,the DM 3m+Anti group and the DM 3m+PBS group increased (t=5.596,3.290,4.286;P=0.000,0.008,0.002).Immunohistochemical staining showed that a small amount of Dl14 was positively expressed in the retinal ganglion cell layer of the normal group;a small amount of VEGFR-2 was positively expressed in the ganglion cell layer and the inner and outer nuclear layers.The positive expression of Dll-4 and VEGFR-2 in retinal vascular endothelial cells of DM 3m group increased significantly.The expression of Dll-4 was significantly decreased in the retinal layers and vascular endothelial cells ofDM 3m+Anti group,while the expression of VEGFR-2 was significantly increased.There was no significant difference between the positive expression of Dll4 and VEGFR-2 in the DM 3m+PBS group and the DM 3m group.The results of real-time PCR showed that the relative expression of Dll-4 and VEGFR-2 mRNA in the DM 3m group was significantly higher than that in the normal group (t=6.705,20.871;P＜0.05).Compared with DM 3m group,the relative expression of Dll-4 mRNA in DM 3m+Anti group decreased,and the relative expression of VEGFR-2 mRNA increased (t=2.681,3.639;P＜0.05).The relative expressions of Dll-4 and VEGFR-2 mRNA in the DM 3m+PBS group and DM 3m group were not statistically significant (t=0.513,0.657;P＜0.05).Conclusions The expression of Dll-4 in retinal vascular endothelial cells is gradually increased during the early retinopathy of DM rats.The expression of Dll-4 is inhibited,the expression of VEGFR-2 is up-regulated,and the plexus edema is alleviated.

Epigenetics palys a important role in many dieases,including diabetic retinopathy (DR).Epigenetic modifications include DNA methylation,histone modifications and deployment of noncoding RNA,which are related to DR.Elucidateing the epigenetic mechanism of DR may provide a new direction for treating diabetes.

OBJECTIVE:To improve HPLC for content determination of ketoprofen in Ketoprofen enteric-coated capsules. METHODS:HPLC method was adopted. The determination was performed on Chiralpak IC column with mobile phase consisted of n-hexane (0.1% TFA)-isopropanol(90:10,V/V)at a flow rate of 0.8 mL/min. The detection wavelength was set at 268 nm,and column temperature was 25 ℃. The sample size was 10 μL. RESULTS:The linear range of ketoprofen were 0.025-0.5 mg/mL(r=0.9998). The limit of quantitation was 1.0 mg/L,and limit of detection was 0.2 mg/L. RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recoveries were 96.36%-100.32%(RSD=1.87%,n=6). CONCLUSIONS:The method is sim-ple,accurate and rapid,and can be used for the content determination of ketoprofen in Ketoprofen enteric-coated capsules.

[Objective] To investigate the mesenchymal stem cells (MSC) in treating acute lung injury (ALI) via ALI mouse model.[Methods] By monoclonal antibody Anti-GD2 of specific antigen ganglioside (GD2) only expressed on MSC as a carrier,new fluorescent molecule probe were synthesized through covalently coupling Anti-GD2 and fluorescent group CyDye mono-reactive NHS Esters (Cy7).Synthetic Anti-GD2-Cy7 and MSC were labeled by the specific binding of antigen and antibody in vitro.Total 84 balb/c male mice were selected and randomly selected 48 mice were divided into three groups:sham group (n =16),MSC+ ALI group (n =16),NS + ALI group (n =16).The lung histopathology and scores,lung W/D ratio and permeability of lung microvasculature were examined at 24 h,48 h after ALI mouse model.Other 36 mice were randomly divided into three groups:normal group (n =12),sham group(n =12),MSC +ALI group(n =12).Labeled MSC-GD2-Cy7 were transplanted into MSC+ALI group and sham group mice via tail vein injection.At 30 min,1 d,3 d,and 7 d post-MSC-GD2-Cy7 injection,the mice were sacrificed after anesthesia and then the lung was removed.Excised lung was detected on small animal fluorescent imager.[Results] Contrast to NS+ ALI group,the lung histopathology and scores,lung W/D ratio and permeability of lung microvasculature of MSC +ALI group were more greatly improved at both 24 h and 48 h.Fluorescent results showed that the signal intensity in thc lung of MSC +ALI group was significantly higher than that of sham group at each time point [(3.37 ± 0.02)× 10-4 vs (2.05 ± 0.04) × 10-4 scaled counts/s;(35.54 ± 0.47)× 10-4 vs (25.29 ± 1.48) × 10-4 scaled counts/s;(11.17 ± 0.75)×10-4 vs (6.09 ± 0.62)× 10-4 scaled counts/s;(3.10 ± 0.14) vs (0.00 ± 0.00)× 10-4 scaled counts/s;all P ＜ 0.05].[Conclusion] Our study showed that a proportion of cells migrated into normal and injured lungs 30 min after cell transplantation,and the cells started to recruit and largely gather in injured lungs at day 1 and persisted to day 7,these results suggest that MSC possess the ability to home into injured tissues.