AIM: To investigate the feasibility to inhibit the expression of MHCⅡ by special siRNA targeting class Ⅱ major histocompatibility complex (MHC Ⅱ) transactivator (CⅡTA), which might regulate MHC Ⅱexpression for suppressing immune rejection. METHODS: Five different siRNA were designed, synthesized and transfected into freshly isolated rat corneal keratocytes. At 24 hours posttransfection, the changes of MHC Ⅱexpression were detected by flowcytometry, and the mRNA abundance of CⅡTA and MHC Ⅱ was measured by FQ-PCR after inducing with recombinant rat interferon-gamma (IFN-?). RESULTS: Different siRNA showed different reduction in MHC Ⅱ and CⅡTA expression compared with the control (P

Objective To investigate the surgical exposure of the subtemporal approach and explore clinical indications using a minimal-access.Methods Ten adult cadaveric heads fixed with formalin were used in this study.The holes with 3.0 cm × 2.5 cm of zygomatic arch vertically were operated on each head with subtemporal approach.During the anatomical procedures,measured the maximal exposure lengths of tentorial margin,posterior cerebral artery,and anterior border of brain stem,vertical distances between highest structure in the field of view and posterior clinoid process,the shortest distances form the zygomatic arch 1/3 to tentorial edge,sulcus lateralis mesencephali and anterior clinoid process; After tentorium of cerebellum was cut,measured the straight distance form the internal carotid artery to the optic nerve and form the posterior communicating artery to the tentorial edge.Results Oculomotor,trochlear nerve,tentorial edge,superior cerebllar artery.,P1-P2 segment of posterior cerebral artery,ventrolateral surface of mesencephalon and pon higher than root of trigeminal nerve,anterior and posterior clinoid process,posterior communicating artery and anterior choroidal artery,and superior portion of pituitary stalk could be observed via subtemporal approach using a minimal-access.Conclusion 1.The subtemporal approach using a minimal-access can protect the superficial temporal artery and the facial nerve branches especially in the process of the flap formation.It does little damage to the temporallis,reduces the invalid exposure of brain tissue,farthest lowers the damage to the scalp,skull and adjacent tissue ; 2.The subtemporal approach using a minimal-access can obtain the exposure rang.It can see the upper pons,petroclival region,tentorial notch area,ventrolateral brain stem.

Purpose To observe therapeutic effect of electroacupuncture Neixiyan (Ex-LE 4 ) and Dubi (ST 35) in treating osteoarthritis of knee joint. Method All the 120 cases were randomly divided into electroacupuncture and control groups, 60 cases in each group,and they were given electroacupuncture and Ritalin slowreleased tablet respectively, and pain, mobility and swelling degree of knee joint were observed before and after treatments. Results In treatment group, the average score increased by 18, while in control group, it increased by 12.33 ( P < 0. 05 ) after treatment. Conclusion Therapeutic effect of electroacupuncture Neixiyan (Ex-LE 4) and Dubi (ST 35) in treating osteoarthritis of knee joint was better than that of administration of Ritalin slow-released tablet.

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective To evaluate the clinical application of SWI and DTI of MRI in the diagnosis and prognosis of diffuse axonal injury (DAI).Methods A retrospective case series study was conducted on the clinical data of 16 patients with DAI admitted from January 2015 to December 2017.There were nine males and seven females,aged (56.3 ± 4.1) years.According to Glasgow Coma Scale (GCS),there were seven patients with 3-8 points,eight with 9-12 points,and one with 13 points.All patients received head CT examination on admission and then received head MRI examination within one week to record the number of lesions on T1WI,T2WI,DWI,and SWI in CT and MRI examination.On the DTI sequence,five regions including the subcortical white matter,the corpus callosum,the thalamus,the cerebellum,and the brain stem were selected for measurement of the apparent diffusion coefficient (ADC) and partial fraction of anisotropy (FA) values.The Glasgow outcome scale (GOS) was evaluated 6 months after injury.The linear correlation between ADC,FA values,GCS,and GOS on admission and after 6 months were analyzed.Results The statistical analysis of CT,T1WI,T2WI,DWI and SWI in 16 patients showed that the detection rates of DAI lesions were 25.6％ (43/168),30.4％ (51/168),44.0％ (74/168),51.8％ (87/168),and 100％,respectively (P ＜0.01).The ADC values of the subcortical white matter,the corpus callosum,the thalamus,the cerebellum,and the brain stem were 0.830 ± 0.148,0.536 ± 0.169,0.838 ± 0.596,0.708 ± 0.157,and 0.713 ± 0.135,respectively,and FA values were 0.487 ± 0.103,0.142 ± 0.040,0.293 ± 0.089,0.212 ± 0.045,and 0.366 ± 0.797,respectively.The GCS on admission was (8.9 ± 3.3)points,and GOS was (4.2 ± 1.0)points six months after injury.The correlation analysis showed that the ADC value and FA value of subcortical white matter and cerebellum were not related to GCS and GOS (P ＞ 0.05).The correlation strength of ADC values in each region with the GCS score in descending order was the thalamus,the corpus callosum,and the brain stem (P ＜ 0.05 or 0.01);for ADC with the GOS score,it was the corpus callosum,the thalamus and the brain stem (P ＜0.05 or 0.01);for FA with GCS and GOS scores,it was thalamus,corpus callosum,and brainstem (P ＜ 0.05 or 0.01).Conclusion The SWI has better sensitivity to detect DAI lesions than CT and conventional MRI sequences.DTI can accurately,objectively and visually detect the integrity of cerebral white matter fibers.Both SWI and DTI can help make early diagnosis and evaluate the prognosis of DAI patients accurately.

Objective To explore the localization and guidance value of infraorbital/maxillary nerve in endoscopic transmaxillary approach, and to provide anatomical data for clinical operation. Methods The eight adult cadaver heads were dissected by means of the endoscopic transmaxillary approach to expose the infraorbital/maxillary nerve, and relevant data were collected. Results In 8 cases, 16 maxillary nerves arose from the trigeminal ganglion of the trigeminal nerve and emerged from the foramen rotundum, transited to the infraorbital nerve at the infraorbital fissure, then traveled in the infraorbital canal and out of the infraorbital foramen. The infraorbital/maxillary nerve could be divided into four segments according to the foramen rotundum, infraorbital groove and infraorbital foramen: the terminal segment, the infraorbital nerve and its terminal branches to the face, distal to the infraorbital foramen; the orbitomaxillary segment and the orbitomaxillary segment of the infraorbital nerve within the infraorbital canal from the infraorbital foramen along the infraorbital groove (length 11.7 ±2.5 mm ), which was readily identified in the roof of the maxillary sinus in all specimens. The pterygopalatine segment, the pterygopalatine segment within the pterygopalatine fossa, which started at the infraorbital groove to the foramen rotundum (length 13.4±2.1 mm); The intracranial segment, the intracranial segment from the foramen rotundum to the trigeminal ganglion(length 15.2±3.9 mm). Conclusion The infraorbital nerve can serve as a anatomical landmark for endoscopic transmaxillary approach to get access to infratemporal fossa, pterygopalatine fossa, trigeminal ganglion and lateral wall of the cavernous sinus.

Objective To investigate the risk factors for contralateral hematoma progression of bilateral chronic subdural hematomas (bCSDHs) after initial unilateral evacuation,and finally develop a prognostic scoring system.Methods Sixty-one patients with bCSDHs underwent initial unilateral evacuation in our hospital from October 2012 to March 2019 were chosen in our study.During follow up,CT examination was used to determine whether the patients had developed eontralateral hematoma.The clinical data of patients from the progressive group and non-progressive group were retrospectively analyzed and compared.Multivariate Logistic regression was used to analyze the independent risk factors for postoperative contralateral hematoma progression.Receiver operating characteristic (ROC) curve was established to predict the progression of contralateral hematoma for each risk factor.A prognostic grading system was developed on the basis of independent risk factors and cut-off value.All patients were scored according to the scoring system and the progression rate of different scores were re-analyzed.Results As compared with the non-progressive group,the progressive group had higher proportions of patients with extensive contralateral hematoma distribution or low density of contralateral hematoma,higher amount of preoperative contralateral hematoma and postoperative hematoma,with statistically significant differences (P＜0.05).Multivariate Logistic regression analysis showed that extensive contralateralhematomadistribution (OR=16.726,95％CI:2.034-137.557,P=0.009),and hematoma volume after contralateral hematoma surgery (OR=1.044,95％CI:1.012-1.078,P=0.007) were independent risk factors for contralateral hematoma progression.ROC curve showed that areas under the curve for contralateral hematoma distribution and hematoma volume after contralateral hematoma surgery were 0.682 and 0.737,respectively.Limited type of contralateral hematoma was set as 0 score,and extensive type was set as one score;hematoma volume after contralateral hematoma surgery＞40 cm3 was set as one score,and that≤40 cm3 was set as 0 score.All patients were scored,and the scoring system was ranged from 0 to 2 scores;the contralateral hematoma progression rate of 0,1,and 2 were 0％,23.81％,and57.69％,respectively,with significantdifferences (P＜0.05).Conclusion Extensive contralateral hematoma distribution and hematoma volume after contralateral hematoma surgery are independent risk factors for contralateral hematoma progression of bCSDHs after initial unilateral evacuation;the prognostic scoring system is simple and practical,which can serve as part of clinical references.

Objective To construct a CaME141G fusion protein-expressing plasmid,and to express,purify,and identify the activity of the recombinant protein. Methods The 141st site of the wild type CaM,E (GAG),was mutated to G (GGG),using site-specific mutagenesis technology. Escherichia coli BL-21 was transformed with the mutant plasmid. The GST-CaME141G fusion protein was mass-cultured and induced for expression. Subsequently,the GST-CaME141G fusion protein was purified using GS-4B beads. PreScission protease was applied to remove the GST,the Bradford method used to determine the concentration of purified protein,and SDS-PAGE used to detect its relative molecular weight and purity. The GST pull-down assay was used to study the protein's biological activity. Results The CaME141G protein was successfully purified at a high concentration and purity. The protein could interact with PreIQ protein fragments from the myocardial CaV1. 2 calcium channel C terminal,in a CaME141G concentration-dependent manner. Therefore,CaME141G has the ability to bind with the CaV1. 2 calcium channel. Conclusion This study successfully constructed a CaME141G fusion protein-expressing plasmid and purified the CaME141G protein. This lays a foundation for regulating the function of CaM mutations in the myocardial CaV1. 2 calcium channel,and for the study of its relationship with diseases of the cardiovascular system.

Objective To investigate the effect of Ghrelin on gastrointestinal motility after traumatic brain injury (TBI).Methods A total of 72 adult male SD rats were randomly divided into sham operation group (n =8),TBI group (n =32) and Ghrelin group (n =32),according to the random number table.In the sham operation group,the scalp was sutured after craniotomy and sterilization,without any strike.In the TBI group,after intraperitoneal anesthesia,the skull was opened and the electric cortical contusion impactor was used to strike the center of bone window at the depth of 3 mm and the rate of 5 m/s.The duration of hitting the lowest point was 200 ms.In the Ghrelin group,20 μg/kg of Ghrelin was injected into the rat via the tail vein 30 minutes after injury.The modified neurologicalseverity score (mNSS),percentage of water content in feces and percentage of gastric contents in body weight at 6,24,48 and 72 hours after operation in each group were measured.The stomach,the small intestine 15 cm from ileocecal junction,ileocecal junction (about 3 cm in the proximal ileal loops,about 3 cm in the distal ileal loops,and 3 cm colon loops) were taken out to prepare the electron microscopy section and observe the microscopic changes of the gastrointestinal mucosa.Results The mNSS in the TBI and Ghrelin groups was higher than that in the sham operation group after 24,48 and 72 hours (P ＜0.01).The mNSS in the TBI group was higher than that in the Ghrelin group after 24,48 and 72 hours (P ＜0.01).In the sham operation group,the intestinal wall was pink.In the TBI group,gastric dilatation and thinner wall with pale or dark red color were seen,and small intestine cavity expansion with dark color and even congestion were observed.There was much mucus in the intestinal wall.The Ghrelin group improved obviously than the TBI group after 6 hours.Compared with the Ghrelin group,the percentage of fecal water content in the TBI group decreased significantly after 24 hours (P ＜ 0.05),and the decrease rate dropped with time.Obvious delayed gastric emptying occurred (P ＜ 0.05),and the percentage of gastric contents in body weight demonstrated downtrend.The changes of gastric mucosa were as follows:the chief cells in the gastric glands were observed 72 hours after TBI in the TBI group,and scattered short microvilli were seen in the cell surface.The cytoplasm protruded into the glandular cavity,and a large number of rough endoplasmic reticulum could be seen in the cytoplasm,with irregular arrangement.Medullary bodies could be seen inside the mitochondria which swelled locally.Abundant endocrine granules were seen in the cytoplasm.Mitochondria were scattered and swollen,and mitochondria cristae became shorter and fewer,which contained medullary bodies.The Ghrelin group improved obviously than TBI group after 72 hours in terms of gastric mucosa changes.With respect to cecum mucosa,in the TBI group 72 hours after TBI,severe edema of the cecum absorption epithelium,obvious dilation of the rough endoplasmic reticulum,expansion of the free water gap inside the cell,and local decrease of the microvilli at the top of the cell were observed.Abundant microvilli were seen in the cecum absorption epithelium and cell top.The connection complex composed of tight connections,intermediate connections,and bridging particle connections could be seen between cells.The Ghrelin group improved obviously than TBI group after 72 hours in terms of cecum mucosa changes.Conclusions Ghrelin can improve gastrointestinal motility and protect gastrointestinal mucosa in rats after TBI.

Objective:To explore the quality differences of systemic biopsy specimens from different regions in prostate biopsy.Methods:The data of 806 patients who underwent transperineal prostate biopsy from May 2013 to December 2020 in Northern Jiangsu People’s Hospital were retrospectively reviewed. The median age of the patients was 72 (66, 77) years old, median PSA was 18.4 (10.3, 34.2) ng/ml, and prostate volume was 43 (32, 56) ml. Tissue quality were graded from low to high as follows. One score means multiple fragments with fragmented tissue ≤5 mm. Two scores means at least one fragment >5 mm and ≤10 mm. Three scores means at least one fragment >10 mm. The prostate specimens fragmentation scores and the length of the specimens in different regions of the prostate were collected to analyze.Results:A total of 806 patients were included in our study. The number of tissues was 8 866, and the mean length of tissues was 1.2 (1.0, 1.5) cm. The tissues of different region were scored according to the scoring criteria, of which 618 (7.0%) prostate tissues were scored as 1 score, 2 720 (30.7%) tissues were scored as 2 scores, and 5 528 (62.4%) tissues were scored as 3 scores. In the prostate apex, tissue quality of 1 score accounted for 11.7%(94/806), 2 scores accounted for 34.7%(280/806), and 3 scores accounted for 53.6%(432/806). While in the prostate base, tissue quality of 1 score accounted for 6.5%(524/8 060), 2 scores accounted for 30.3%(2 449/8 060), and 3 scores accounted for 63.2%(5 096/8 060)( H=35.850, P<0.05). The mean length of the prostate apical tissue was 1.0 (0.8, 1.3) cm, which was significantly shorter than prostate basal tissue of 1.2(1.0, 1.5) cm ( Z=-11.353, P<0.05). Conclusions:In transperineal prostate biopsy, the apical tissue was more fragmented and shorter, prostate apex should be concerned.