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Objective:To compare the effects of different intraocular infusion solutions on histology and function of retina.Methods:Human corneal endothelial cells (HCEC), human retinal pigment epithelium (HRPE) cells and rat retinal ganglion cells (RGC) were divided into normal control group, balanced saline solution (BSS) group and compound electrolyte intraocular irrigating solution (CEIIS) group, and the cells were cultured in 10% DMEM/F12 medium, BSS and CEIIS for 12, 24 and 48 hours, respectively, according to grouping.The proliferation absorbance value of cultured cells was measured by cell counting kit-8 (CCK8) method.The expression of apoptosis related proteins in cultured cells was detected by cellular immunofluorescence staining.The cell apoptosis rate and cell cycle were measured by flow cytometry.The mitochondrial damage was detected by lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) quantitative detection kit.Fifteen New Zealand white rabbits were randomly divided into control group ( n=3), BSS group ( n=6) and CEIIS group ( n=6). The left eyes were taken for vitrectomy and different intraocular perfusion fluids were used during vitrectomy according to grouping.The retinal function of operative eyes was measured by flash electroretinogram (ERG) before operation and 24 hours after operation, and the structural changes of each layer of retina were detected by optical coherence tomography (OCT). The early apoptosis of retinal cells was detected by TUNEL staining.The expressions of cytochrome C and bax protein in retina were detected by immunohistochemical staining.The ultrastructural changes of retina were observed under a transmission electron microscope.The use and care of animals complied with the ARVO statement.This study protocol was approved by an Ethics Committee of Peking University People's Hospital (No.2019PHE059). Results:The three kinds of cultured cells in BSS and CEIIS groups were damaged in various degrees.With the extension of culture time, proliferated cells were decreased and the number of apoptotic cells was increased.Compared with the BSS group, cultured cells in the CEIIS group were dense and in orderly arrangement with uniform morphology and size.The apoptosis rates of HRPE cells and RGC in the BSS group were (37.157±6.918)% and (29.993±12.330)%, respectively, which were significantly higher than (4.163±1.310)% and (6.337±1.903)% in the CEIIS group ( P=0.003, 0.045). There was no significant difference in G0/G1+ S phase ratio of HCEC and HRPE cells among the normal control group, BSS group and CEIIS group (HCEC: F=2.226, P=0.189; HRPE: F=2.634, P=0.151), and the proportion of G2/M division arrest phase of RGC in the BSS group was significantly higher than that in the normal control group and CEIIS group ( P=0.047, 0.024). The proliferation absorbance values of HCEC, HRPE cells and RGC in the CEIIS group were significantly higher than those in the BSS group at each culture time point (all at P<0.05). The fluorescence intensity of cytochrome C, bax, caspase-3 and caspase-9 proteins in the BSS group was stronger than that in the normal control group and CEIIS group, and the fluorescence intensity of bcl-2 was weaker than that in the CEIIS group, and the fluorescence intensity of zonula occluden-1 (ZO-1) was weaker than that in the normal control group and CEIIS group.The release level of LDH in the BSS group was significantly higher than that in the CEIIS group at different time points (all at P<0.001). After 48 hours of culture, the release level of SDH in the BSS group was significantly higher than that in the CEIIS group ( P<0.05). No retinal histological abnormalities was found through OCT examination of rabbit eyes after vitrectomy in the two groups, but transmission electron microscopy showed that there were different degrees of loose arrangement of retinal photoreceptor cells, a large number of photoreceptor outer membrane discs falling off and vacuolar degeneration in the two groups, especially in the BSS group.TUNEL staining showed that the apoptotic cells were mainly located in the inner nuclear layer and RGC layer.The number of apoptotic retinal cells was (135.2±22.8)/high-power field of vision in the BSS group, which was significantly higher than (81.3±17.7)/high-power field of vision in the CEIIS group ( t=4.175, P=0.002). Full field flash ERG showed that the amplitudes of scotopic 3.0 ERG a- and b-wave in the CEIIS group after operation were significantly lower than those before operation, but the differences were not statistically significant (all at P>0.05). The amplitudes of scotopic 3.0 ERG a- and b-wave in the BSS group after operation were significantly lower than those before operation ( P=0.026, 0.010). Conclusions:In vivo and in vitro research results show that compared with BSS, there were few apoptotic cells in retinal tissue after vitrectomy perfused by CEIIS.
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Objective To explore the expression of N-myc downstream regulated gene 1 (NDRG1) in gastric carcinoma and the relationship with clinicopathological parameters and prognosis. Methods The expression of NDRGI was detected by immunohisto chemistry in formalin-fixed and paraffin-embedded sections with a total of 220 specimens including 110 gastric carcinoma and 110 corresponding paraneoplastic tissue. The correlation between clinicopathological parameters and the expression of NDRG1 in gastric carcinoma were also analyzed. Results Low expression of NDRG1 was detected in most gastric carcinoma sections. Among the gastric cancer tissues, NDRG1 protein expression was significantly lower in tumors with more advanced pathological stage, local tumor invasion and lymphatic metastases. There was no significant difference in sex, age, tumor differentiation and gross types of the tumor. The 1-, 3- and 5 year survival and disease free survival in patients with low NDRG1 protein expression was 84.2%, 53.9%, 21.1%, and 60.5%, 31.6%, 19.7%, respectively, which was signifivantly poorer when compared with patients with high NDRG1 protein expression. Conclusion The expression of NDRG1 is low in the majority of patients with gastric carcinoma, which was in a close relationship with advanced stage, local invasion and lymphatic metastases of gastric carcinoma. NDRG1 may be a candidate metastasis suppressor gene.
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Objective To investigate The effect of Heat shock protein 70(HSP70) antisense oligonucleotides (ASO)to bladder carcinoma in mouse loaded with tumor.Methods The 40 mice loaded with tumor subcutaneously were established by cultured BIU-87 cells,and divided into 4 groups randomly when the subcutaneous neoplasms grew to about 100 mm3,namely,HSP70 mRNA ASO plus mitomycin C(MMC)group;HSP70 mRNA ASO group;MMC and blank control.HSP70 mRNA ASO were injected into neoplasms,10mmg/kg weight,twice every week,and MMC 0.1mg/kg weight,twice every week,and the above schemes were replaced with normal saline to blank.The neoplasms were peeled off,photograghed and weighed in 30 days.HSP70 expressions were examined with reverse transcription polymerase chain reaction (RT-PCR),mierovaseular density(MVD)was evaluated by immunohis to chemical staining and the tumor cells apoptosis was detected by terrainal deoxynucleotidyl transferase(TdT)-mediated dUTP-biotin nick end labeling technique (TUNEL).Results The tumor inhibition rate in ASO+MMC surpassed 50%.more than ASO or MMC respectively,and the differences were significantly(P＜0.05).The ASO and MMC exceeded blank group respectively(P＜0.05).The ASO was the same as the MMC(P＞0.05).The apoptotic index(AI)in ASO+MMC surpassed the other three groups (P＜0.05).The difference between ASO and MMC was not significant (P＞0.05),while the A1 of ASO or MMC was more than blank respectively(P＜0.05).The results of MVD were in accordance with the above results.Conclusion The injection of HSP70 mRNA ASO in tumor locally can inhibit neoplasm growth,and this effect might correlate with the inhibition of apoptosis and microvascular forming resulting from the ASO.
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<p><b>OBJECTIVE</b>To explore the pathogenic changes of myocardial apoptosis in heart hypertrophy during hypertension and evaluate the anti-apoptosis effect of Valsartan.</p><p><b>METHODS</b>Thirty spontaneously hypertensive rats (SHRs) were divided into two groups: 15 treated with Valsartan (20 mg x kg(-1) x d(-1)) (SHR + Valsartan group), the others with placebo (SHR + placebo group), with 15 normal Wistar rats as control. Systolic blood pressure was measured by the tail-cuff method. The observation period was from 8 to 16 weeks of age. Cardiac apoptosis was evaluated by a Terminal Deoxynucleotidyl Transferase-Mediated dUTP-biotin Nick End Labeling (TUNEL) assay.</p><p><b>RESULTS</b>Mean blood pressure values were 127 +/- 2 mm Hg in controls, 163 +/- 6 mm Hg in the SHR + Valsartan group and 193 +/- 7 mm Hg in the SHR + placebo group at 16 weeks of age, whereas the blood pressure in 8-week-old SHR and Wistar rats were 175 +/- 3 mm Hg and 125 +/- 5 mm Hg, respectively. The ratio of the heart weight over body weight declined in Wistar (3.07 +/- 0.03 mg/g) and SHR + Valsartan groups (3.22 +/- 0.19 mg/g) compared with the SHR + placebo group (4.02 +/- 0.31 mg/g) (P < 0.05). The density of TUNEL-positive cells in Wistar and SHR +/- Valsartan groups was 23.3 +/- 3.3 nuclei/HPF and 35.0 +/- 1.3 nuclei/HPF, both of which were significantly less than that of the SHR + placebo group (116.7 +/- 11.3 nuclei/HPF).</p><p><b>CONCLUSIONS</b>In response to chronic pressure overload, cardiomyocyte-specific apoptosis contributes to the transition from compensatory hypertrophy to decompensation. Apoptosis may be effectively inhibited by Valsartan in the early stage of hypertension.</p>
Subject(s)
Animals , Rats , Antihypertensive Agents , Pharmacology , Apoptosis , Cardiomegaly , Pathology , Hypertension , Pathology , Myocardium , Cell Biology , Rats, Inbred SHR , Rats, Wistar , Tetrazoles , Pharmacology , Valine , Pharmacology , ValsartanABSTRACT
ObjectiveTo investigate the clinicopathological features of the needle biopsies of prostatic lesions and to evaluate the value of 34?E12 immunostaining for the diagnosis of prostatic carcinoma.MethodsThe clinical data,levels of serum PSA and HE slides of 103 cases of the needle biopsies of prostatic lesions were reviewed.Immunohistochemical stains for 34?E12 were performed.ResultsThe morphological features revealed 36 cases of benign prostatic hyperplasia(BPH),9 cases of low grade prostastic intraepithelial neoplasia (PIN I)and 34 cases of prostatic carcinoma(PC),24 cases of high grade PIN(PINⅡ and Ⅲ) have been suspected of carcinoma.With immunohistochemical stain,all cases of BPH were strongly reactive for 34?E12 while 32 cases of prostatic carcinoma were negative;14 of 24 cases of high grade PIN were negative for 34?E12 and could be diagnosed as PC.The rest 10 of high grade PIN were interruptedly positive in the basal layer for 34?E12.In the 30 cases of prostastic carcinoma,serum PSA levels of 25 cases was over 10 ng/ml and over 50 ng/ml in 15.ConclusionsThe pathologic diagnosis should based on gland architectures and cytologic features.34?E12 negative expression and elevation of serum PSA are also the important criteria for the diagnosis of prostatic carcinoma.
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Objective To evaluate the relationship between Helicobacter pylori (Hp) infection and pathohistological presentation of gastric and deuodenal mucosa.Methods The gastritis biopsies were taken through endoscopy from each gastric body,antrum and duodenum in 60 patients with gastritis and 22 patients with duodenal ulcer respectively for the examination of histology.Result The positive rate of Hp was nearly 100% in active chronic gastritis which was significantly higher than that of active duodenitis.The total detecting rate of Hp infection and incidence rate of active inflammation in antrum were obviously higher than that of gastric body and duodenal bulb.The detecting rate of Hp infection of duodenal bulb in 22 cases of duodenum ulcer was 54% which was significantly lower than that of antrum.Conclusion The Hp infection is a very important pathogenic factor of active inflammation in gastric mucosa on the basis of which the severe inflammation of mucosa and ulcer development.Therefore the early elimination of Hp is very important.
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Objective To investigate the expression and pathological diagnostic utility of ?-methylacyl-coenzyme A racemase(P504S)in prostate cancer(PC). Methods The specimens of 42 cases of PC,18 cases of prostatic intraepithelial neoplasia(PIN)and 25 cases of benign prostatic hyperplasia(BPH)tissues were immunohistochemical stained with P504S,34?E 12 and vascular endothelial growth factor(VEGF)antibody,respectively. Their staining extent and intensity were analyzed and compared. Results Of the 42 PC cases,33(79%)showed positive P504S staining with moderate to strong staining intensity, which was significantly stronger than that of PIN and BPH cases,while adjacent normal glands were all negative. Weakly positive P504S staining was found in 6(33%)of 18 PIN cases and 2(8%)of 25 BPH cases,while it was negative in others.The specimens of BPH cases had a complete basal cell layer and were positive for 34?E 12 ;while 40 cases of PC had no basal cell layer and only 2 cases had discontinuous basal cell layers with weak staining.The basal cell layers of PIN cases were positive for 34?E 12 and the staining was a little weaker than that of BPH cases.All the specimens of BPH,PIN and PC cases showed positive staining for VEGF,the extent and staining intensity had no significant difference among these groups. Conclusions P504S has high sensitivity and good specificity in the diagnosis of PC,and is helpful for differentiating the suspected cases when combined with 34?E 12.
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Objective To clarify the role of a cell cycle regulator, cyclin D1 and CDK6 in patients with gastric carcinoma. Method Tissue samples from 48 patients with gastric carcinoma were included in the current study. Expression levels of cyclin Dl and CDK6 in samples of normal mucosa and tumor tissue were analyzed by Western blot. Result Overexpression of cyclin Dl and CDK6 protein were demonstrated in 58% and 69% of gastric cancer tissues, respectively. Several clinicopathologic parameters, including depth of tumor invasion, pathologic lymph node status and tumor stage ( P
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Objective To investigate the relationship between HPV infection and human colorectal carcinogenesis. Methods Colorectal carcinoma specimens from 72 Chinese patients were studied. DNA extracted from colorectal tissue was screened for HPV L1 by polymerase chain reaction (PCR), HPV subtype 6,11, 16, and 18 were detected by PCR using specific primers and in situ hybridization using specific probe. Results Twenty-four specimens out of 72 (33%) colorectal cancer were HPV L1 positive. The normal colorectal mucosa was HPV L1 negative. The location, infiltration and metastasis of colorectal carcinoma were all significantly related with HPV infection. The predominant HPV subtype was HPV 16,which was found in 58% of all HPV-positive colorectal carcinoma. Conclusion The presence of HPV DNA suggests that HPV may be involved in the carcinogenesis of colorectal carcinoma. HPV infection is closely related with the malignant potential of colorectal cancer.
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Objective To explore the mechanism of HPV infection in carcinogenesis and progression of colon cancer. Methods Human colon cancer cells, HCT116 (with wild-type p53) and SW480 (with mutant-type p53), were transfected by HPV16 E6E7 oncogenes using a recombinant adeno-associated virus vector system. The transfection efficiency was determined by flow cytometry. The expression of HPV16 E6 genes was determined by Western blot. The cell proliferation and cell cycle was studied by MTT method and flow cytometry. Results Western blot confirmed the expression of E6 gene in colon cells that were infected by rAAV-E6E7. The population doubling time of HCT116 cell, which was more than 48 hours at control group, decreased to 33 hours. HPV16 E6E7 increased cell percentage of S phase and decreased cell percentage of G0/G1. The population doubling time of SW480 cell was 77.06% decreased and the OD540 was 47.18% increased with interference of HPV16 E6E7 gene. Conclusion HPV16 E6E7 oncogene precipitates the proliferation and positively controls cell cycle of HCT116 and SW480 human colon cancer cells. HPV infection may closely relate to the carcinogenesis and progression of colon cancer.