ObjectiveTo investigate the change in the activity of hepatitis B virus （HBV）-specific CD8⁺ T cells after pegylated interferon-α-2b （PEG-IFN-α-2b） therapy in HBeAg-negative patients with chronic HBV infection. MethodsA total of 53 HBeAg-negative patients with chronic HBV infection who attended The First Affiliated Hospital of Xinxiang Medical University and Tangdu Hospital of Air Force Mdical University from April 2020 to June 2022 were enrolled and treated with PEG-IFN-α-2b （180 μg/week， subcutaneous injection） antiviral therapy. The study endpoint was HBsAg clearance （course of treatment<48 weeks） or 48 weeks （course of treatment≥48 weeks）. Peripheral blood mononuclear cells were isolated at baseline and study endpoint， and peripheral blood T cell counts were measured. Enzyme-linked immunospot assay was used to measure the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ. A total of 17 HLA-A^*02-restricted patients were selected， and CD8⁺ T cells were purified to establish direct- and indirect-contact co-culture systems for HBV-specific CD8⁺ T cells and HepG2.2.15 cells. The level of lactate dehydrogenase in supernatant was measured to calculate the mortality rate of HepG2.2.15 cells， and the levels of HBV DNA， cytotoxic molecules， and cytokines in supernatant were also measured. Flow cytometry was used to measure the expression of apoptosis ligands， and the cytotoxicity of HBV-specific CD8⁺ T cells was evaluated. The independent samples t-test or the paired t-test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test or the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsThe HBsAg clearance rate at study endpoint was 30.19% （16/53）. There were no significant differences in peripheral blood T cell counts （CD3⁺， CD4⁺， and CD8⁺ T cells） between baseline and study endpoint （P>0.05）. At study endpoint， there was a significant increase in the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ （U=177.50， t=11.90， U=186.50， all P<0.001）， and the patients with HBsAg clearance had a significantly higher frequency of such HBV-specific CD8⁺ T cells than those without HBsAg clearance （U=120.50， t=2.73， U=121.50， all P<0.01）. In the direct- and indirect-contact co-culture systems at study endpoint， HBV-specific CD8⁺ T cells induced a significant reduction in HBV DNA in the supernatant of HepG2.2.15 cells （all P<0.001） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （all P<0.05）； in the direct-contact co-culture system， HBV-specific CD8⁺ T cells induced significant increases in the mortality rate of HepG2.2.15 cells （13.62%±3.27% vs 11.39%±2.40%， t=2.27， P=0.030） and the secretion of perforin and granzyme B （t=72.50， U=52.50， both P<0.05）. In the direct- and indirect-contact co-culture systems， compared with HBV-specific CD8⁺ T cells from the patients without HBsAg clearance， the HBV-specific CD8⁺ T cells from patients with HBsAg clearance had a significantly greater reduction in HBV DNA （P<0.05） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （P<0.05）. ConclusionPEG-IFN-‍α‍-2b therapy can help to achieve a relatively high HBsAg clearance rate in HBeAg-negative patients with chronic HBV infection， and the activity of HBV-specific CD8⁺ T cells is significantly enhanced， which is closely associated with HBsAg clearance.

Objective To predict the potential geographic distribution of Oncomelania hupensis in Yunnan Province using random forest (RF) and maximum entropy (MaxEnt) models, so as to provide insights into O. hupensis surveillance and control in Yunnan Province. Methods The O. hupensis snail survey data in Yunnan Province from 2015 to 2016 were collected and converted into O. hupensis snail distribution site data. Data of 22 environmental variables in Yunnan Province were collected, including twelve climate variables (annual potential evapotranspiration, annual mean ground surface temperature, annual precipitation, annual mean air pressure, annual mean relative humidity, annual sunshine duration, annual mean air temperature, annual mean wind speed, ≥ 0 ℃ annual accumulated temperature, ≥ 10 ℃ annual accumulated temperature, aridity and index of moisture), eight geographical variables (normalized difference vegetation index, landform type, land use type, altitude, soil type, soil textureclay content, soil texture-sand content and soil texture-silt content) and two population and economic variables (gross domestic product and population). Variables were screened with Pearson correlation test and variance inflation factor (VIF) test. The RF and MaxEnt models and the ensemble model were created using the biomod2 package of the software R 4.2.1, and the potential distribution of O. hupensis snails after 2016 was predicted in Yunnan Province. The predictive effects of models were evaluated through cross-validation and independent tests, and the area under the receiver operating characteristic curve (AUC), true skill statistics (TSS) and Kappa statistics were used for model evaluation. In addition, the importance of environmental variables was analyzed, the contribution of environmental variables output by the models with AUC values of > 0.950 and TSS values of > 0.850 were selected for normalization processing, and the importance percentage of environmental variables was obtained to analyze the importance of environmental variables. Results Data of 148 O. hupensis snail distribution sites and 15 environmental variables were included in training sets of RF and MaxEnt models, and both RF and MaxEnt models had high predictive performance, with both mean AUC values of > 0.900 and all mean TSS values and Kappa values of > 0.800, and significant differences in the AUC (t = 19.862, P < 0.05), TSS (t = 10.140, P < 0.05) and Kappa values (t = 10.237, P < 0.05) between two models. The AUC, TSS and Kappa values of the ensemble model were 0.996, 0.954 and 0.920, respectively. Independent data verification showed that the AUC, TSS and Kappa values of the RF model and the ensemble model were all 1, which still showed high performance in unknown data modeling, and the MaxEnt model showed poor performance, with TSS and Kappa values of 0 for 24%(24/100) of the modeling results. The modeling results of 79 RF models, 38 MaxEnt models and their ensemble models with AUC values of > 0.950 and TSS values of > 0.850 were included in the evaluation of importance of environmental variables. The importance of annual sunshine duration (SSD) was 32.989%, 37.847% and 46.315% in the RF model, the MaxEnt model and their ensemble model, while the importance of annual mean relative humidity (RHU) was 30.947%, 15.921% and 28.121%, respectively. Important environment variables were concentrated in modeling results of the RF model, dispersed in modeling results of the MaxEnt model, and most concentrated in modeling results of the ensemble model. The potential distribution of O. hupensis snails after 2016 was predicted to be relatively concentrated in Yunnan Province by the RF model and relatively large by the MaxEnt model, and the distribution of O. hupensis snails predicted by the ensemble model was mostly the joint distribution of O. hupensis snails predicted by RF and MaxEnt models. Conclusions Both RF and MaxEnt models are effective to predict the potential distribution of O. hupensis snails in Yunnan Province, which facilitates targeted O. hupensis snail control.

Objective To analyze the values of serum tumor necrosis factor-α stimulated gene 6 protein (TSG-6) and type ⅩⅥ collagen (col-16) in predicting the prognosis of patients with ulcerative colitis (UC). Methods A total of 160 patients with UC were enrolled as UC group, another 160 volunteers were selected as control group, and the levels of serum TSG-6 and col-16 were compared between the two groups. According to the modified Mayo scoring criteria, patients in the UC group were divided into mild group (n=63), moderate group (n=52), and severe group (n=45), and the levels of serum TSG-6 and col-16 were compared among the three groups. Based on the colonoscopy follow-up results, patients in the UC group were divided into good prognosis group (n=121) and poor prognosis group (n=39), and the levels of serum TSG-6 and col-16 were compared between the two groups. Spearman correlation analysis was used to explore the correlations of serum TSG-6 and col-16 with the modified Mayo score. Logistic regression analysis was used to investigate the impact of serum TSG-6 and col-16 on the prognosis of patients in the UC group. Receiver operating characteristic (ROC) curve was used to assess the values of serum TSG-6 and col-16 in predicting the prognosis of patients in the UC group. Results The levels of serum TSG-6 and col-16 in the UC group were significantly higher than those in the control group (P < 0.01). The levels of serum TSG-6 and col-16 in the moderate and severe groups were significantly higher than those in the mild group, and the levels of serum TSG-6 and col-16 in the severe group were significantly higher than those in the moderate group (P < 0.01). The levels of serum TSG-6 and col-16 in the poor prognosis group were significantly higher than those in the good prognosis group (P < 0.01). Spearman correlation analysis showedthat serum TSG-6 and col-16 were positively correlated with the modified Mayo score (r=0.691, P < 0.05; r=0.635, P < 0.05). Logistic analysis results showed that serum TSG-6 and col-16 were risk factors for poor prognosis (P < 0.001). ROC curve analysis showed that the areas under the curve for predicting poor prognosis in patients with UC were 0.773 and 0.765 for serum TSG-6 and col-16, respectively. Conclusion The levels of serum TSG-6 and col-16 are abnormally elevated in patients with UC, and there is a certain correlation of the two indexes with the severity and prognosis of disease, and the two indexes can be used as predictors of poor prognosis.

ObjectiveTo evaluate the efficacy of sphenopalatine ganglion stimulation with acupuncture for moderate-to-severe perennial allergic rhinitis.MethodsA total of 50 patients were recruited into a sphenopalatine ganglion stimulation group and a routine acupuncture group according to order of presentation, with 25 in each group. The sphenopalatine ganglion stimulation group received sphenopalatine ganglion stimulation with filiform needle, 1-2 sessions/week for 4 weeks. The routine acupuncture group received traditional acupuncture, withyingxiang(LI 20),yintang(GV29),fengchi(GB20),fengfu(GB16),zusanli(ST36) as the maln points, andyingxiang(LI 20),yintang(GV29),fengchi(GB20),fengfu(GB16),zusanli(ST36) as the adjunct points, 1-2 points from both the maln and adjunct points in each session, 2 sessions/week for 4 weeks. The nasal symptom score (2004 version), the total nasal symptom score (TNSS) and the total non-nasal symptom score (TNNSS) were used to evaluate symptom improvement. The Rhinoconjunctivitis Quality of Life Questionnalre (RQLQ) was used to assess the patients’ quality of life. The time to symptom alleviation, duration of symptom alleviation in every session and the recurrence duration during 1 month after the treatment were compared between the two groups.Results After the treatment, the score of the nasal symptom score (99.74 ± 31.89vs.196.83 ± 31.22;t=-4.912,P=0.001), TNSS (33.63 ± 12.37vs.71.82 ± 19.21;t=-3.463,P=0.003), TNNSS (33.63 ± 12.37vs.71.82 ± 19.21,t=-3.463,P=0.003) in the sphenopalatine ganglion stimulation were significant lower than those in the routine acupuncture group. Compared with the routine acupuncture group, the time to symptom alleviation was significant shorter (13.85 ± 4.21 minvs.45.63 ± 7.87 min;t=-1.763,P=0.008), while the duration of symptom alleviation was significant longer (37.92 ± 9.94 hvs.3.35 ± 1.23 h;t=7.637,P<0.01) after each session in the sphenopalatine ganglion stimulation group. Four weeks after the treatment, RQLQ score in the sphenopalatine ganglion stimulation group was significant lower than that in the routine acupuncture group (8.48 ± 3.71vs.37.68 ± 12.46;F=-7.312,P<0.01). The recurrence duration during 1 month after the treatment in the sphenopalatine ganglion stimulation group was significant longer than that in the routine acupuncture group (4.12 ± 2.15 dvs.23.53 ± 4.63 d;t=-8.879,P=0.003).ConclusionSphenopalatine Ganglion stimulation is superior to routine acupuncture in treatment of patients with moderate-to-severe perennial allergic rhinitis.

Objective To retrospectively analyze computerized tomography (CT) imaging features of colorectal cancer with chronic schistosomiasis in order to improve the diagnostic accuracy.Methods Eighty patients whose diagnosis was pathologically confirmed as colorectal cancer with schistosomiasis were collected from Tongji Hospital and Kunshan Chinese Medicine Hospital from January 2007 to December 2012.All the patients underwent abdominal plain CT and contrast-enhanced CT scan.The lesion location,morphology,size,calcification features,enhancement patterns and cancer metastasis were evaluated and compared by two radiologists who were blind to the diagnosis.Twenty colorectal cancer cases without schistosomiasis from the same area were also collected as controls.Results CT imaging showed that the tumors all occurred in the colon and rectum in 80 patients,mainly in colon descendens,colon sigmoideum and rectum.The lesion was characterized by irregular bowel wall thickening with soft tissue masses,and the average length of impaired intestine was (21.35 ± 4.50) cm.The lesions were solitary in 68 cases (85.0％) and were multifocal in 12 cases (15.0％).Linear,spotty and small patchy calcifications were seen in all the patients,with margins unclear in 59 patients (73.8％) and margins clear in 21 patients (26.2％).Early phase enhancement was seen in 68 cases (85.0％),of which 45 cases (56.2％) had homogenous enhancement and 23 cases (28.8％) had heterogeneous enhancement.Late phase enhancement was seen in 73 cases (91.2％) and necrosis was hardly seen.Only two cases (2.5％) had liver metastasis without lymphatic metastasis.All the 20 colorectal cancer cases without schistosomiasis presented with single lesion,which was characterized by irregular bowel wall thickening and local ulcer.The average length of impaired intestine was (6.90±3.40) cm.No calcification was seen in these lesions.All cases had early phase lesion with remarkable heterogeneous enhancement with low density necrotic area.The late phase enhancement was decreased.Four cases (20.0％) had liver metastasis.Compared to cases without schistosomiasis,cases with schistosomiasis tended to have multifocal (x2=100,P=0.000) and longer lesion (t=45.506,P=0.000),and more calcification (x2=100,P=0.000).The early phase heterogeneous enhancement was less frequent (x2 =88,P=0.000) and late phase enhancement was more frequent (x2=100,P=0.000).The liver metastasis rate was significantly lower (x2 =8.688,P =0.014).Conclusions The CT imaging of colorectal cancer with schistosomiasis is characterized by calcifications inside the tumor with obscured margins and multiple intestinal segments involvement.Hematogenous metastasis and lymphatic metastasis are rarely seen.

Objective To label neural stem cells (NSCs) with superparamagnetic iron oxides (SPIO) and to explore the tropism of NSCs after transplantation into the hippocampus of APP/PS1 AD mice by MRI.Methods NSCs from C57BL/6 mouse were cultured and identified.Feridex and Poly-L-Lysine were added into the medium to be co-cultured to make magnetic labeled NSCs and transmission electron microscopy was used to identify the iron particles in NSCs.Transgenic (tg) and wild-type (wt) mice at 12 months of age were divided into three groups: SPIOs labeled NSCs group (A and C),unlabeled NSCs group(B).Feridex-labeled NSCs were migrated into the hippocampus of APP/PS1 AD mice to monitor in vivo by MRI.After 1,2,4 and 6 weeks,the mice were sacrificed and their brain tissues were sectioned to investigate the migration of SPIO labeled NSCs and compared with MRI.Results NSCs of C57BL/6 mice were cultured successfully.Transmission electron microscope showed visible iron granules in cytoplasm.MRI detection of labeled cells: T2WI and T2* WI showed remarkable low signal intensity at the hippocampus injection points 1 week after transplantation,particularly on T2* WI.Area of low signal intensity enlarged increasingly along the injection points after 2 weeks.At 4 weeks,area of low signal intensity spread throughout the hippocampus,but intensity shadowed Six weeks later,low signal intensity almost disappeared.There was no obvious low signal change in unlabeled cell transplantation group.For wt mice,size and location of low signal did not appear obvious change at all designated time points.Prussian blue positive cells were observed in the hippocampus,indicating that NSCs labeled with SPIO could survive,migrate and differentiate in the brain of the APP/PS1 AD mice.Changes of pathology were well correlated with the area where a signal intensity loss was observed in MRI 1,2,4 and 6 weeks after transplantation Conclusions Diffuse migration of transplanted NSCs labeled with SPIO is observed in the hippocampus in APP/PS1 tg mice,and MRI technique is an ideal method for tracking labeled stem cells after grafting in vivo.

Objective To explore the effect of neural stem cell(NSCs) transplantation on proton magnetic resonance spectroscopy (1H-MRS) and the behavior in APP/PS1 double transgenic AD mice.Methods NSCs from C57BL/6 mice were cultured and amplified.APP/PS1 double transgenic AD mice (n=30) aged 12 months were used as the study group,and mild-type mice (n=15) were used as the control group(group C).Animals in the study group were randomly divided into two subgroups:one receiving NSCs (group A) and the other receiving PBS transplantation (group B) in bilateral hippocampal CA1 of the AD model mice.Animals in the group C were not treated.1 H-MRS and Morris water maze (MWM) were performed before transplantation and 4 weeks after transplantation,and compared with the histopathological results.Results 1H-MRS showed that there was no significant change in NAA/Cr(1.01±0.08 and 1.03±0.05) and mI/Cr (0.69±0.05 and 0.71±0.06) ratios between group A and group B before transplantation (P＞ 0.05),but the changes were significant compared with the group C (NAA/ Cr:1.21±0.05; mI/Cr:0.58±0.06) (P＜0.05).Four weeks after transplantation,NAA/ Cr ratio(1.18± 0.09) was increased and mI/Cr ratio (0.53±0.04) was decreased in group A.The difference was significant compared with the group B at the same time points (P＜0.05).MWM showed the escape latency in group A was significantly shorter than that in group B after transplantation (P＜0.05).In addition,group A also showed an exclusive preference for the target quadrant,and spent more time ((35.21±5.44) s) in the 3rd quadrant compared with group B (P＜0.05).For number of platform crossings,similar results were also shown (5.75± 3.23).Nissl's staining showed that the number of neurons in the hippocampal area increased more significantly in group A than those in group B(P＜0.05).Conclusion NSCs transplantation can improve spatial learning and memory via neurons regeneration in APP/PS1 double transgenic AD mice,and 1H-MRS is able to display intracranial metabolite changes after NSCs transplantation.

ObjectiveInvestigate the value and feasibility of image registration with reverse rotation in lateral cerebral DSA.Methods ( 1 ) Experimental study:the target images were subtracted directly,and subtracted again after reverse rotation.Software of registration and subtraction with reverse rotation edited by the author utilizing Visual Basic.The function of the automatic angle detection by the software were evaluated to see whether it detected the angle of line.The subtraction function of DSA by the software was evaluated.(2) Clinical retrospective study:the untreated mask and target inages of 15 patients with motion along vertical axis during lateral cerebral DSA were uploaded to the software.The target images were processed with and without the software to get two sets of images.( 3 ) Evaluation:four experienced radiologists read and compared the two sets of the images,and graded their findings.Results ( 1 ) The automatic detection by the software suggested that the target images should be rotated counterclockwise 1.3°. The subtraction result of the software was satisfactory.(2)In the 15 sets of images,there were only three sets of images deemed optimal after traditional subtraction.After reverse rotation,artifacts were significantly reduced and the image sharper. There were ten cases with significant artifacts after traditional subtraction,and those images were sharper and showed more peripheral vessels after reverse rotation. The traditional subtraction images of two sets could not be interpreted,the reverse rotation registration images reached the diagnostic quality.(3)Subjective evaluation: there were more information and less noise and distortion in the registration images with reverse rotation than in the traditional subtraction. But the image resolution decreased slightly after reverse rotation registration.ConclusionThe registration of digital angiography with reverse rotation can improve the image quality in lateral cerebral DSA.

Objective To study the asymmetry of the soft tissue thickness of upper eyelids in Shanghai female undergraduates undergoing double eyelid surgery.Methods Data were collected in 565 female students from Shanghai universities.Before surgery,oblique sagittal view images of the upper eyelids by 3.0T MRI were obtained and the following parameters were measured: soft tissue thickness at upper tarsal plate margin and upper central tarsal plate,sub-eyebrow and intra-orbital septum fat pad areas,and whole upper eyelid soft tissue areas.The resected orbicularis and intra-orbital septum fat were weighed by a highly accurated electronic balance.The eyelid asymmetry index (EAI) was calculated.Results The MRI (U/U2) central tarsal plate soft tissue thickness were:4.46±0.90 in the right side and 3.78±1.01 in the left; the intra orbital septum fat areas were:172.33±49.29 in the right and 136.34±37.42 in the left; the whole tissue areas were: 697.13±146.99 in the right and 500.66±158.87 in the left (P＜0.01 for all).The weight of the resected orbicularis oculi muscle and intra orbital septum fat pad were (0.18±0.05) g and (0.17±0.06) g for the right side,and (0.15±0.04) g and (0.06±0.05) g for the left side (P＜0.01 for all),respec tively.The orbicularis EAI was 0.17±0.06,and the intra orbital septum fat EAI was 0.41 ±0.08.Conclusions The asymmetric phenomenon of the upper eyelids' soft tissue thickness is commonly found in the Shanghai female undergraduates,and the main manifestation is that orbicularis and intra orbital septum fat in right side are thicker than that in the left.

Objective To explore the value of 1H-MRS on the evaluation of Alzheimer's disease (AD) with neural stem cells (NSCs) transplantation in an APP-PS1 double transgenic (tg) AD mouse model.Methods NSCs from C57BL/6 mice were cultured and amplified.APP-PS1 tg mice (n =30) aged 12 months were used as the study group,and mild-type mice (n =15) were used as the control group.Animals in the study group were randomized into two subgroups,the AD mice in one subgroup received NSCs transplantation (NSCs group) and in another subgroup received phosphate buffer saline (PBS,PBS group)in bilateral hippocampal CA1.Animals in the control group were not treated.Using a 7.0 T high-fieldstrength MR imager,1H-MRS was performed before and 6 weeks after transplantation to measure the area under the peak of n-acetyl aspartate (NAA),glutamate (Glu),myo-inositol ( mI),choline (Cho) and creatine (Cr) in the hippocampal area,NAA/Cr,Glu/Cr,mI/Cr and Cho/Cr ratio were calculated and compared with histopathological results (including Nissl's staining and electron microscope examination).Comparisons among NSCs,PBS and control groups were conducted by one-way ANOVA.Results NSCs from C57BL/6 mice were cultured successfully. Before transplantation,the mean NAA/Cr,Glu/Cr and mI/Cr in NSCs,PBS and control groups were 0.89 ± 0.05,0.88 ± 0.04 and 1.15 ± 0.05,0.40 ± 0.03,0.39 ± 0.03 and 0.45 ± 0.05,0.67 ± 0.05,0.67 ± 0.05 and 0.52 ± 0.04,respectively,and differences were statistically significant (F =148.918,7.529,59.468,P ＜ 0.01 ). There were no significant differences in NAA/Cr,mI/Cr and Glu/Cr ratios between NSCs and PBS groups before transplantation (t =0.147,0.096,0.207,P ＞ 0.05 ),but the differences were significant compared with the control group (t =0.255,0.467,0.171 and t =0.269,0.527,0.151,P ＜0.05).Six weeks after transplantation,the mean NAA/Cr,Glu/Cr and mI/Cr in three groups were 1.13 ±0.07,0.86 ±0.05 and 1.14 ±0.05,0.45 ± 0.04,0.38 ± 0.02 and 0.44 ± 0.03,0.58 ± 0.04,0.67 ± 0.04 and 0.53 ± 0.04,respectively,and differences were statistically significant ( F =112.092,23.076,44.367,P ＜ 0.01 ).NAA/Cr and Glu/Cr ratios were increased and mI/Cr was decreased in NSCs group,and the difference was significant compared with PBS group at the same time point ( t =0.271,0.071,0.089,P ＜ 0.05 ).There were no significant differences in NAA/Cr and Glu/Cr ( t =0.013,0.012,P ＞ 0.05 ),but there was a significant difference in mI/Cr between NSCs and control groups ( t =0.046,P ＜ 0.05).There were no significant differences in Cho before and after transplantation among the three groups (P ＞ 0.05 ). Nissl's staining showed that the number of neurons in the hippocampal area increased more significantly in tg mice receiving NSCs than that without receiving NSCs.Electron microscopy showed that most hippocampal NSCs in NSCs group were morphologically normal with abundant organelles,while hippocampal NSCs in PBS group were swollen with sparse synapses.Conclusion 1H-MRS is able to display intracranial metabolite changes before and after NSCs in APP-PS1 double transgenic AD mice and has an applicable value in evaluating the therapeutic effect of NSCs on AD.