Objoctive:To investigate the expression of transforming growth factor-β(TGF-β)isoforms in colonic mucosa of ulcerative colitis(UC),and its role in the incidence of UC thereof.Methods:The expression of TGF-β isoforms was detected in colonic mucosa by immunohistochemical Envision method in UC patients,and the patients with irritable bowel syndrome (IBS)were used as controls.The relationship was analyzed between the expression of TGF-β isoforms and the degree of severity and extent of the disease.Results:There were statistical differences in the expression of TGF-β1 and TGF-β2 between the active stage UC,remission stage UC and IBS groups(P ＜ 0.01).The expression was significantly higher in active stage UC group than that in remission stage UC and 1BS groups(P＜ 0.01).The expression was significantly higher in remission stage UC group than that in IBS group(P ＜ 0.01).However,there was no statistical difference in the expression of TGF-β3 between active stage UC,remission stage UC and IBS groups(P＞ 0.05).The expression of TGF-β1 and TGF-β2 had positive correlation with the degree of sevefity(P ＜ 0.05).However,the expression of TGF-β3 had no linear correlation with the degree of severity(P ＞ 0.05).There was no linear correlation on the expressions of TGF-β1,TGF-β2 and TGF-β3 with extent of the disease(P ＞ 0.05).The degree of severity had positive correlation with extent of the disease(P ＜ 0.05).Conclusion:The expressions of TGF-β1 and TGF-β2 were enhanced in colonic mucosa of UC,and were correlated with the pathogenesis of UC.The expressions of TGFβ1 and TGF-β2 may serve as markers for assessing disease activity of UC.

The goal of this work was to explore the prospect of standardized application of an in-vitro bioactivity assay for recombinant human follicle-stimulating hormone based on a reporter gene. The relative accuracy, intermediate precision, linearity and applicable range of the method were validated according to the General Rules of Chinese Pharmacopoeia 2020 edition Volume IV (9401). Three laboratories used this method to determine the in-vitro biological activities of six batches of drug product and three batches of drug substance manufactured by two different companies. The consistency of the potency determined by three laboratories, the intra-laboratory precision and inter-laboratory precision were analyzed. The method was optimized during the collaborative validation. The results of method validation meet the requirements of the General Rules of Chinese Pharmacopoeia 2020 edition Volume IV (9401). Aiming to resolve the problems found in the collaborative validation, the medium for cell seeding, the pre-diluted buffer solution of standard and sample, and the means of removing and discarding supernatant after stimulation were optimized. After optimization, there was no significant difference in the bioactivity among the different laboratories (P > 0.05), indicating statistical equivalency. Intra-laboratory and inter-laboratory precision were good and the geometric coefficient of variation (GCV%) were both less than 15%. In conclusion, the reporter gene assay has good intra-laboratory repeatability and inter-laboratory reproducibility and is suitable for analyzing recombinant human follicle-stimulating hormone drug product and drug substance by different manufacturers. It is expected to be used as a standardized method for the determination of the in-vitro bioactivity of such products.

Objective To investigate the clinical value of mean channel of neutrophil volume (MNV), mean channel of neutrophil conductivity (MNC) and mean channel of neutrophil scatter (MNS) in predicting acute bacterial infection.Methods Peripheral blood samples from 112 patients with positive blood cultures for bacteria,70 healthy subjects and 45 non-infectious subjects with high white blood cell count(WBC) were studied using the Coulter LH 750 hematology analyzer.MNV, MNC, MNS and neutrophil volume distribution width (NDW) were retrospectively analyzed and compared with total WBC, percentage of neutrophils,neutrophil left-shift and CRP.112 blood bacterial infections were grouped according to WBC count (A:WBC ＜11.0×109 /L;B:11.0×109/L≤WBC＜15.0×109 /L;C:WBC≥15.0×109 /L) and neutrophil rate (NE ＜ 0.85 and NE ≥ 0.85 ).Results MNV and NDW increased significantly in septic patients (154.17 ± 10.08,24.36 ± 4.14 ) compared with those of healthy control group (142.09 ± 4.13,19.04 ± 1.97) and non-infectious patients with high WBC group ( 150.63 ± 8.14,20.19 ± 4.73 ).There was statistically significant difference (F value were 20.738 and 28.190 respectively,P ＜ 0.01 ). On the contrary, MNS decreased significantly in septic patients (137.15 ± 7.61 ) compared with that of healthy group (144.51±4.36) and nonspetic patients with high WBC group (142.45±7.11) ,there was significant statistical difference (F=5.217,P＜0.01).The MNV, NDW and MNS of A group were 148.09±5.76,22.39±1.97,140.07±6.11 respectively.The MNV, NDW and MNS of B group were 152.83±5.75,24.14±1.35,141.44±5.35 respectively.The MNV, NDW and MNS of C group were 164.28±6.49,29.42±5.93,134.27±9.61 respectively. There was statistically significant difference compared with healthy group (F value were 24.720,31.642,7.931, P ＜ 0.01).The MNV, NDW and MNS in the group with NE ＜0.85 were 149.17±9.06,22.59±2.73,141.19±4.34 respectively.The MNV, NDW and MNS in the group with NE≥0.85 group were 159.03±10.23,27.64±4.51,135.62 ± 8.95 respectively.There was statistically significant difference compared with healthy group ( F value was 23.970,51.309,19.792,P＜0.01).With a cut-off of 150 for the MNV, a specificity of 90% and sensitivity of 70% were achieved.NDW was associated with neutrophil left-shift (r=0.33,P＜0.01).With a cut-off of 23 for the NDW, a specificity of 100% and a sensitivity of 72% were achieved.The sensitivity of the MNV and NDW was better than total white blood cells count (with a cut-off≥ 11.0×109 /L, the sensitivity was 57% ), percentage of neutrophils( with a cut-off≥0.85, the sensitivity was 44% ) and neutrophil shift to left ( with a cut-off ＞5%, the sensitivity was 66% ) and CRP (with a cut-off ≥10 mg/L, the sensitivity was 65% ).Conclusions The MNV and NDW of the neutrophil can reflect the morphologic change of neutrophil sensitively and specificialy in acute infection. As quantitative, objective and more sensitive parameters, MNV and NDW may have a potential role for predicting the acute bacterial infection.

ObjectiveTo compare tongue positions at rest between normal occlusion people and patients with anterior cross-bite malocclusions,and to explore whether there is any abnormality in the tongue position of the anterior cross-bite malocclusions.Methods Twenty adults of normal bite (8 males and 12 females,with an average age of 23.8 years) and 20 adults of anterior cross-bite (1 1males and 9 females,with an average age of 24.6 years) were involved in this study.The position habits of tongue were investigated by a questionnaire.The tongue positions,which were enhanced by applying barium to tongue and palatal surface,were compared by cephalometric roentgenogram.Two groups were compared by t-test to determine the variation of anterior cross-bite. Results In normal occlusion group,the dorsum of tongues was sealed with all palatal surfaces except uvula site.In anterior cross bite group,tongues were far from palate,and were difficult to lift.The profile of tongue showed standing upright forward and up in normal occlusion group,while inclining back and low in anterior cross-bite group.The tops of tongue dorsum sites were at the border of soft and hard palate in normal occlusion group.While in anterior cross bite group,the tops of tongue dorsum sites were at the border of uvula and soft palate.In anterior cross-bite group,the tongue-to palate distance was bigger than that of normal occlusion group (P＜0.01).However,both length and height of the tongue had no significant differences between two groups (P＞0.05).ConclusionsIn anterior cross-bite patients,tongue positions were lower than that of normal occlusion people.

Objective To study the effect of radiotherapy and prognostic factors for patients with post-operative loco-regional recurrence of esophageal carcinoma. Methods From 2000 to 2005,91 patients with esophageal carcinoma who developed post-operative loco-regional recurrence were analyzed retrospectively. The interval between surgery and recurrence was 1 -35 months, with a median interval of 11.1 months. There were 4 patients with anastomosis relapse, 6 with anastomosis and mediastinal lymph nodes relapse,4 with abdominal lymph node relapse, 20 with supra-clavicular lymph node relapse, 34 with mediastinal lymph node relapse, and 23 with mediastinal and supra-clavicular lynph node relapse. There were 56 and 35 patients who received conventional and three-dimensional conformal radiation therapy respectively, with a total radiation dose of 50 -70 Gy. Sixty-eight patients received adjuvant chemotherapy.Results The follow-up rate was 95%. The 1-,2-and 3-year overall survival rates were 52%, 20% and 14%, respectively. In univariate analysis, the recurrence time (P = 0. 001), postoperative stage (P =0. 000), radiation dose (P =0. 001) ,overall response rate (P =0. 000) ,T stage (P =0. 028), and N stage (P = 0. 003) were related with prognosis. Multivariate analysis showed that the recurrence time (P =0. 014), postoperative stage (P = 0. 006), radiation dose (P = 0. 009), overall response rate (P = 0. 000)were independent prognostic factors for survival. Conclusions Radiotherapy may improve the survival of esophageal carcinoma patients with postoperative recurrence. Patients with long recurrence-free time, early stage, high response rates, and radiation dose of higher than 60 Gy have better prognosis.

Objective:To observe the effect of CD40 siRNA on the changes in kidney,urinary protein and complement C3 of MRL/Lpr mice and explore its therapy on lupus nephritis.Methods: 16 female MRL/Lpr mice were randomly divided into control group,empty vector group,CD40-siRNA1 group and CD40-siRNA2 group.The vectors expressing siRNA against CD40 were injected by tail veil into MRL/Lpr mice,while MRL/Lpr mice in control group and empty vector group were injected with the same dose of PBS and pGFP-V-RS vector respectively.There were six times injection and every one day.The 24 hours urine output was gathered 24 hours before mice were killed.Fourteen days following administration,these mice were killed,tissue sections of kidney were observed if the signal of siRNA were expressed in kidney.The expression levels of CD40 mRNA and protein in kidney tissue of MRL/Lpr mice were detected by RT-PCR and immunohistochemistry methods respectively.At the same time,the pathological changes of the kidney were observed by haematoxylin-eosin ( HE) staining method.The 24 h urinary protein content was detected using the method of coomassie brilliant blue and the expression levels of complement C3 in serum were detected by Immunoturbidimetric assays.Results:The vector of CD40-siRNA was expressed in kidney of MRL/Lpr.The expression levels of CD40 mRNA and protein in kidney were lower in CD40-siRNA1 group and CD40-siRNA2 group than control group and empty vector group on the 14th day after last injection ( P<0.05).The inflammatory cells infiltration of kidney and some glomerular volume were significantly reduced in CD40-siRNA1 group and CD40-siRNA2 group than control group and empty vector group.The renal tubular swelling was alleviated in CD40-siRNA1 group and CD40-siRNA2 group than control group and empty vector group.The levels of 24 hours urinary protein were lower and the levels of complement C3 were higher in CD40-siRNA1 group and CD40-siRNA2 group than control group and empty vector group ( P<0.05). Conclusion:CD-40 siRNA can suppress the expression levels of CD40 mRNA and protein and decrease inflammatory cells infiltration in kidney of MRL/Lpr.Meanwhile after suppressing expression of CD40 mRNA and protein, it can reduce the content of 24 hours urinary protein and elevate the level of complement C3 in serum,which CD-40 siRNA can delay progress of the disease and protect kidney,so that it has therapy effect on lupus nephritis.

Objective:To study the effect of the Shengmai injection combined with norepinephrine on cellular immunity in rats with multiple organ dysfunction syndrome .Methods:40 healthy male rats were randomly divided into the control group ,the Shengmai injection group ,the norepinephrine group and the Shengmai injection combined with norepinephrine group of 10 rats in each group after MODS rat model were established by intraperitoneal injection of zymosan .The plasma level of TNF-α, IL-1β, IL-12 and IL-2 were detected by ELISA ,and the protein expression of caspase-3,caspase-9 and caspase-8 in spleen were tested by IHC during 6 h,24 h,48 h.And the lymphocyte number of apoptosis and the ratio of Th 1/Th2 were tested by flow cytometry.Results: ① Compared with the control group, the plasma concentrations of TNF and IL-1βwere not significant increased in the Shengmai injection group , the norepinephrine group and the Shengmai injection combined with norepinephrine group ,as the extension of treatment time ,the plasma concentration of TNF and IL-1βwere significant lower in treatment groups than control group .While the plasma concentration of IL-12 and IL-2 were significantly increased in treatment groups , and the combined treatment group was more obvious .② Compared with control group,the protein expression of caspase-3,caspase-9 and caspase-8 in spleen were decreased in the Shengmai injection group , the norepinephrine group and the Shengmai injection combined with norepinephrine group ,and the combined treatment group was more obvious.③The lymphocyte number of positive apoptosis were significantly decreased , and the Th1/Th2 ratio were increased in the Shengmai injection group ,the norepinephrine group and the Shengmai injection combined with norepinephrine group ,and the combined treatment group was more obvious .Conclusion: The Shengmai injection combined with norepinephrine can reduce the inflammatory reaction and improve the immune function in the rat with multiple organ dysfunction .

ObjectiveTo investigate portal hemodynamic features and its changes following devascularization in cirrhotics. MethodsHemodynamics of portal trunk (PT), right anterior branch (RAB) and splenic vein (SV), including maximum cross-sectional velocity(CS-Vmax), flow volume and congestion index(CI), were assessed in 69 cirrhotics and 46 normal volunteers by using a recently-developed color Doppler velocity profile(CDVP). Of 28 patients undergone devascularization procedure, portal hemodynamics were studied and compared before and after operation. ResultsCSVmax of PT and RAB was significantly lower in cirrhotic group than normal group;PT and SV flow volume and the ratio of SV to PT flow volume(SV/PT)were significantly greater in cirrhotic group compared with those of normal group;CI of PT, RAB and SV was significantly higher in cirrhotic group than normal group. Postoperative PT flow volume was significantly reduced( P ＜0.01),and the reduction was closely related to preoperative SV flow volume( r = 0.65, P ＜0.001). CS-Vmax and flow volume in RAB were decreased significantly following operation( P ＜0.01), and the reduction of RAB flow volume was highly related to preoperative RAB flow volume( r =0.74, P ＜0.001). After operation, free portal pressure(FPP)was declined by (6 ± 5)cmH2O[ (0.59 ± 0.49)kPa] ( P ＜ 0.001 ). There were no significant changes in CS-Vmax, CI in PT and CI in RAB following operation. ConclusionIn cirrhotics with portal hypertension, portal venous system coexists the elevated vascular resistance and hyperdynamics, but with different redorainance at different portion. SV hyperdynamics is the main source of increased portal blood flow. Devascularization procedure could markedly relieve portal hyperdynamics by elimination of SV inflow, which is one of the main mechanisms in obmining therapeutic goal. But the operation has no favorable effects on the increased portal resistance,and portal perfusion to the liver would be further declined after relieving portal hyperdynamics, which is unfavorable to maintenance of liver function.

OBJECTIVETo understand the possible pathogenesis of sporadic multiple system atrophy (MSA).

METHODSThe immunoreactivity and ultrastructural features of glial cytoplasmic inclusions (GCIs) in 12 autopsy patients with MSA and 4 normal control groups were studied. All regional sections from each subject were evaluated with HE staining, Klüver-Barrera (KB), Holzer's, modified Gallyas-Braak's (GB) methods and immunohistochemical staining with alpha-synuclein and ubiquitin antibodies. Pontine white matter with abundant GCIs from case 1 was examined, using conventional electron microscopy, Gallyas-Braak's electron microscopy and immunoelectron microscopy.

RESULTSThe presence of GCIs as constantly demonstrated in all MSA patients. Strong alpha-synuclein immunoreactivity was observed in all of the ubiquitinated GCIs. However, the density of alpha-synuclein positive GCIs differed from case to case, and there was no relationship between the density of GCIs and age, sex, or MSA subtype. Ultrastructural features indicated that argyrophilic granule-associated filaments of about 25 nm in diameter were the predominant constituents of GCIs, and the anti alpha-synuclein antibody selectively labeled in these filaments. No GCIs and alpha-synuclein immunoreaction were found in control brain tissues.

CONCLUSIONSGCI was a pathognomonic change in sporadic MSA patients. Accumulation of alpha-synuclein in GCIs may occur during the early stags of MSA. Seletcive alpha-synuclein positive abnormal microtubules in GCIs therefore play an important role in the pathogenesis of MSA.

Aged ; Aged, 80 and over ; Female ; Humans ; Immunohistochemistry ; Inclusion Bodies ; ultrastructure ; Male ; Microscopy, Immunoelectron ; Middle Aged ; Multiple System Atrophy ; etiology ; metabolism ; pathology ; Nerve Tissue Proteins ; analysis ; Neuroglia ; ultrastructure ; Synucleins ; alpha-Synuclein

BACKGROUND: As a pain-related peptide, substance P is closely related with sustained pain in low back and lower extremities caused by lumbar spinal stenosis. However, the expression of substance P in the rat with cauda equina compression rat is not reported.OBJECTIVE: To investigate the content of substance P in spinal cord of rat with lumbar spinal stenosis at different stages.DESIGN: A randomized and controlled experimental study.SETTING: The Orthopedic Department of the First Affiliated Hospital of China Medical University.MATERIALS: The trial was conducted in the Experimental Animal Center of China Medical University within the period from February 2003 to December 2004. Thirty mature male SD rats were randomly and averagely assigned into five groups. One group served as control and in the other four experimental groups the content of substance P of the rats were examined in the 2nd, 4th, 8th and 12th respect week respectively.METHODS: The model of lumbar spinal stenosis was established by compressing cauda equina in rats. The spinal cords were harvested in the 2nd,4th, 8th and 12th respect week respectively for immunohistochemical determination of the expression of substance P (SP) in the posterior horn.MAIN OUTCOME MEASURES: The expression of substance P in the posterior horn at different times. Deeply-stained neurons in the 2nd week group. The mean positive neuron group varied within a certain range and were less than that of the 2nd week group. But they were all more than that of the controls[(5.08 ±0.41),(4.02±0.35), (2.58±0.33), (3.45±0.22) vs(0.85±0.23), t=6.85,5.90,3.49,6.85,P ＜0.05 or P ＜0.01].CONCLUSION: The expression of substance P increases when the cauda equina is compressed. Whether this is because the harmful stimuli increases SP expression and the latter in turn protects spinal cord by reducing the edema of spinal cord warrants more studies.