Objective To study the expression of heat shock protein 47 (HSP47) and its correlation to collagen deposition in pathological scar tissues. Methods The tissues of normal skin(10 cases), hypertrophic scar(19 cases), and keloid(16 cases) were obtained. The expression ofHSP47 was detected by immunohistochemistry method. The collagen fiber content was detected by Sirius red staining and polarization microscopy method. Results Compared with normal skin tissues(Mean IOD 13 050.17±4 789.41), the expression of HSP47 in hypertrophic scar(Mean IOD -521 159.50±272994.13) and keloid tissues(Mean IOD 407 440.30±295 780.63) was significantly high(Plt;0.01). And there was a direct correlation between the expression of HSP47 and the total collagen fiber content（r＝0.386，Plt;0.05）. Conclusion The HSP47 is highly expressed in pathological scartissues and it may play an important role in the collagen deposition of pathological scar tissues.
ObjectiveTo explore the predictive value of preoperative serum heat shock protein 90α (HSP90α) level in combination with the prognostic nutritional index (PNI) for patients with hepatocellular carcinoma (HCC) after transarterial chemoembolization (TACE). MethodsThe HCC patients confirmed by histopathological examination and underwent TACE at Guigang People’s Hospital from January 2022 to June 2023 were as the observation group, the healthy individuals who underwent physical examinations during the same period and same hospital as the control group. The blood before treatment and on the day of the physical examination was collected to detected the HSP90α and albumin levels, as well as lymphocyte count. The PNI was calculated [PNI=albumin (g/L)+5×lymphocyte count (×109/L)]. The clinical outcome (tumor progression or death) was observed within one year after TACE treatment, those without tumor progression or death were defined as a good prognosis, while those with tumor progression or death were defined as a poor prognosis. Using the multivariate unconditional logistic regression analysis to identify the risk factors affecting the poor prognosis for HCC patients, and the receive operating characteristic (ROC) curve to evaluate the predictive value of serum HSP90α level in combination with PNI in distinguishing prognosis after TACE treatment.ResultsIn this study, there were 178 cases in the observation group and 100 cases in the control group. The serum HSP90α level (μg/L) in the observation group was higher than that in the control group (96.40±33.57 vs. 52.19±22.13, t=3.191, P<0.001), and the PNI value was lower than that in the control group (43.70±5.24 vs. 56.46±6.86, t=–16.144, P<0.001); Within one year after TACE treatment, there were 70 patients with poor prognosis and 108 patients with good prognosis. The serum HSP90α (μg/L) level in the patients with poor prognosis was higher than that in the patients with good prognosis (117.33±29.48 vs. 82.83±28.84, t=7.726, P<0.001), and the PNI was lower than that in the control group (40.49±4.18 vs. 45.78±4.80, t=–7.548, P<0.001). The multivariate unconditional logistic regression analysis found that the probabilities of incidence of poor prognosis after TACE treatment were higher in the patients with Chinese liver cancer staging Ⅲa–Ⅲb stage [reference: Ⅰ–Ⅱa stage, OR (95%CI)=5.332 (1.058, 26.875), P=0.043] and increased age and HSP90α level [OR (95%CI)=1.100 (1.025, 1.180), P=0.008; OR (95%CI)=1.049 (1.029, 1.070), P<0.001] , as well as decreased PNI value [OR (95%CI)=0.772 (0.686, 0.869), P<0.001]. The area under the ROC curve after TACE treatment in the HCC patients by serum HSP90α level in combination with PNI was 0.878 [95%CI=(0.820, 0.922)] in differentiating poor prognosis or not. ConclusionThe analysis results of this study suggest that preoperative serum HSP90α level in combination with PNI has a higher predictive value for prognosis of HCC patients after TACE treatment.
ObjectiveTo determine the effects of different volume fluid resuscitation on intestinal injury and the permeability of intestine in hemorrhagic shock rats.
MethodsSprague-Dawley male rats(n=72) were randomly equally divided into 4 groups after the model establishment of blood pressure-controlled hemorrhage, 45, 30, and 15 mL/(kg·h) of fluid resuscitation were performed in high dosage of resuscitation(HLR), moderate dosage of resuscitation(MLR), and low dosage of resuscitation(LLR) group respectively, but rats of Sham group didn't accept fluid resuscitation. After resuscitation, ten centimeters ileum was harvested for testing intestinal permeability. Then 6 rats of each group were sacrificed at 24, 48, and 72 hours after fluid resuscitation respectively. Over the specified time interval, blood was collected for testing levels of lactic acid and plasma tumor necrosis factor-α(TNF-α). The ileums of 3 resuscitation groups were obtained for testing the ratio of wet weight to dry weight and observing the histological changes.
ResultsAfter resuscitation, the intestinal permeability was higher in HLR group(P<0.05). At 3-8 hours after resuscitation, rats of Sham group were all died, and the other rats of 3 groups were all alive. The level of plasma lactic acid was lower in LLR group than those of other 2 groups at 24 hours(P<0.05). The levels of TNF-α were higher in HLR group than those of other 2 groups at 24, 48, and 72 hours(P<0.05), and at 48 hours, level of TNF-α in LLR group was lower than MLR group(P<0.05). At 24 hours after resuscitation, ratio of intestinal wet weight to dry weight in LLR group was the lowest, and HLR group was the highest(P<0.05). According to the histopathology, intestinal injuries of the 3 groups were tend to be remission with the time, and at 48 and 72 hours after resuscitation, intestinal villus of LLR group appeared to be normal.
ConclusionLimited fluid resuscitation of 15 mL/(kg·h) could not only decrease the levels of lactic acid and TNF-α, but also moderate the intestinal permeability and the intestinal injury in early stage after shock and surgery.
Objective
To investigate the expression of induced heat shock protein (HSP) 70 in ratprime;s retinal neurons (RNs) and Muuml;ller cells, and evaluate the protective effect of HSP 70 on RNs injured with glucose deprivation and glutamate.
Methods
Ratprime;s RNs and Muuml;ller cells cultured in vitro were treated with heat shock (42℃ for 1 hour), and duration of the expression of HSP70 was detected by immunocytochemical techniques. Viability of the cells was measured by methyl thiazolyl tetrazolium (MTT) chromatometry after incitant toxic injury with glucose deprivation (0.56 mmol/L glucose for 6 hours) and glutamate (100 mu;mol/L for 6 hours). Simultaneously, the expression was interdicted by HSP70.
Results
Hypereffective expression of HSP70 was found in cultured RNs and Muuml;ller cells after heat shock. The viability of RNs pretreated by heat shock after injured with glucose deprivation and glutamate significantly increased which could be interdicted by HSP70 antibody.
Conclusion
Hypereffective expression of HSP 70 may be induced by heat shock, which enhances the ability of tolerance of RNs to the incitant toxic injury by glucose deprivation and exitotoxicity.
(Chin J Ocul Fundus Dis, 2005,21:110-113)
Objective To analyze the protective effects of heat-shock response on the retinae of the rats after retinal ischemic reperfusion injury.Method Twenty Wistar rats (20 eyes) were divided into 4 groups: intracameral perfusion group (group P), intracameral perfusion after quercetin injection group (group P+Q), intracameral perfusion after heat shock group (group P+H), and in tracameral perfusion after quercetin injection and heat shock group (group P+Q+H ). According to the standard program established by International Society for Clinical Visual Electrophysiology, we recorded the results of the dark-adapted electroretinogram (D-ERG ),oscillatory potentials (OPs),and light-adapted ERG (L-ERG) of the rats with intraocular hypertension after induced by heat shock response. The expressions of HSP 70 of the rats in all groups were observed by Western blotting.Results The expression of HSP 70 of the rats in group P+H was the highest in all groups, but the expressions of HSP70 in group P+Q and P+Q+H were inhibited significantly. The amplitudes of a and b wave of ERG and O2 wave of OPs decreased, and the delitescence of them were delayed significantly in rats after intracameral perfusion. The amplitude of b wave of D-ERG and O2 wave of OPs in group P+H were higher than which in group P. Zero hour after perfusion, the amplitudes of all waves in group P+H increased significantly (Plt;0.05). Twenty-four hours after perfusion, the retinal functional resumption of the rats in group P+H was better than which in group P. In group P+Q and P+Q+H, the delitescences of all waves of ERG and O2 wave of OPs were the longest and the amplitudes were the lowest, and some waves even disappeared.Conclusions The heat-shock response may improve the recovery ability of the retinal cells after injury of ischemic reperfusion.(Chin J Ocul Fundus Dis，2003，19：117-120)
Objective Extracorporeal shock wave (ESW) can promote angiogenesis and tissue repair. To investigate the influence of ESW therapy on the histological features of diabetic chronic wounds and wound healing. Methods Ninety-six male Sprague Dawley rats with weight (220 ± 20) g were divided into 3 groups (n=32): diabetic control group, ESW treatment group, and normal control group. The diabetic rats were prepared in diabetic control group and ESW treatment group by intraperitoneal injection of Streptozotocin (60 mg/kg). Then a circular full-thickness skin wound of 1.8 cm in diameter was made at the back of diabetic rats to establish the diabetic chronic wound model, and the same wound was made in normal control group. In ESW treatment group, ESW (0.11 mJ/mm2, 1.5 Hz energy, and 500 pulses) was applied to treat the wound at 1 day after wounding; in two control groups, no ESW treatment was given. The wound healing and histological changes were observed by HE and Masson staining at 3, 7, and 14 days after treatment; and the cell proliferation, angiogenesis, and collagen deposition were observed by CD31 and proliferating cell nuclear antigen (PCNA) immunohistochemical staining. Results The wound closure rate in diabetic control group was lower, and the healing time was significantly longer than those in normal control group (P lt; 0.05); at 3, 7, and 14 days after treatment, the inflammatory cell infiltration in wound tissue was obvious, and the relative area density of collagen fibers, wound microvessel density (MVD), and the relative density of PCNA-positive cells were significantly lower than those in normal control group (P lt; 0.05). The wound healing time was significantly shorter and the wound closure rate was significantly higher in ESW treatment group than those in the diabetic control group (P lt; 0.05). At different time points in ESW treatment group, the inflammatory cells signficantly reduced, while the relative area density of collagen fibers, MVD, and relative density of PCNA-positive cells significantly increased when compared with those in diabetic control group (P lt; 0.05). No significant difference in MVD and relative density of PCNA-positive cells was found between ESW treatment group and normal control group (P gt; 0.05). Conclusion Low-energy ESW treatment can inhibit the local inflammatory response, promote cell proliferation, increase angiogenesis and collagen deposition, and enhance granulation tissue formation, and so it can promote chronic wound healing in diabetic rats.
Extracorporeal shock wave treatment is capable of providing a non-surgical and effective treatment modality for patients suffering from osteoarthritis. The major objective of current works is to investigate how the shock wave (SW) field would change if a bony structure exists in the path of the acoustic wave. Firstly, a model of finite element method (FEM) was developed based on Comsol software in the present study. Then, high-speed photography experiments were performed to record cavitation bubbles with the presence of mimic bone. On the basis of comparing experimental with simulated results, the effectiveness of FEM model could be verified. Finally, the energy distribution during extracorporeal shock wave treatment was predicted. The results showed that the shock wave field was deflected with the presence of bony structure and varying deflection angles could be observed as the bone shifted up in the z-direction relative to shock wave geometric focus. Combining MRI/CT scans to FEM modeling is helpful for better standardizing the treatment dosage and optimizing treatment protocols in the clinic.
ObjectiveTo observe the expression of heat shock protein 47 (HSP47) and transforming growth factor-β2(TGF-β2) in vitreous specimens and epiretinal membranes of patients with proliferative vitreoretinopathy diseases.
MethodsVitreous specimens and epiretinal membranes were obtained from 48 patients (48 eyes) with proliferative vitreoretinopathy (PVR) and 50 patients (50 eyes) with proliferative diabetic retinopathy (PDR). Vitreous specimens and internal limiting membranes were collected from 20 patients (20 eyes) with idiopathic macular hole (IMH) as control group. The expression of HSP47 and TGF-β2 in the vitreous specimens was evaluated using enzyme linked immunosorbent assay. The expression of HSP47, TGF-β2, typesⅠandⅢcollagen in epiretinal membrane and internal limiting membrane specimens were observed for immunohistochemical staining method. The correlation between the positive expression of HSP47 and TGF-β2, typesⅠandⅢcollagen in epiretinal membrane specimens of patients with PVR and PDR were analyzed.
ResultsThe expression of HSP47 in vitreous specimens of patients with PVR, PDR and IMH were (212.35±23.32), (231.30±26.79), (171.06±28.91) pg/ml, respectively. The expression of TGF-β2 in vitreous specimens of patients with PVR, PDR and IMH were (1919.96±318.55), (1939.39±177.57), (1194.61±234.20) pg/ml, respectively. The expression of HSP47, TGF-β2 in the vitreous specimens of patients with PVR and PDR were significantly increased compared with patients with IMH and the difference was statistically significant (F=12.952, 34.532;P < 0.01). The epiretinal membrane of patients with PVR and PDR showed markedly increased expression of HSP47, TGF-β2, typesⅠandⅢcollagen in the cytoplasm and extracellular matrix. The expression of HSP47 and typeⅢcollagen was negative and the expression of TGF-β2 was weakly positive and the expression of typesⅠcollagen was positive in internal limiting membrane of patients with IMH. The expression of HSP47, TGF-β2, typesⅠandⅢcollagen in the epiretinal membrane of patients with PVR and PDR were significantly increased compared with patients with IMH and the difference was statistically significant (F=13.469, 18.752, 12.875, 20.358; P < 0.01). The expression of HSP47 was positively correlated with the positive expression of TGF-β2, typesⅠandⅢcollagen in epiretinal membrane specimens of patients with PVR (r=0.475, 0.556, 0.468; P < 0.05) and PDR (r=0.484, 0.589, 0.512; P < 0.05).
ConclusionsThis study showed increased consistent expression of HSP47 and TGF-β2 in vitreous and epiretinal membrane specimens of patients with PVR and PDR. Both HSP47 and TGF-β2 were expressed in the cytoplasm and extracellular matrix. HSP47 and TGF-β2 may be involved in the pathological process of PDR and PVR by promoting collagen synthesis.
Objective To investigate the role and mechanism of heat shock protein 60 (HSP60) in induction of murine skin allograft tolerance. Methods At the age of 8-12 weeks, inbred female BALB/C (H-2d) mice (n=45) and CBA/N (H-2k)mice (n=15) were used as transplantation donors and C57BL/6 (H-2b) mice (n=60) as recipients. Recipients C57BL/6 (H-2b) mice were randomized into 4 groups (n=15). In group A, 1 cm × 1 cm Wolfe-Krause skin graft was excised from the back of BALB/C (H-2d) mice and hypoderma was scraped off aseptically, and then transplanted to the back of C57BL/6 (H-2b)mice. The method of skin transplantation in the other 3 groups was the same as to group A. In group B, C57BL/6 (H-2b) mice were treated with imcompleted Freund’s adjuvant (IFA) administration into the back 2 weeks before transplantation of BALB/C (H-2d) mice skin. In group C, C57BL/6 (H-2b) mice were administered HSP60 emulsified in IFA into the back 2 weeks before transplantation of BALB/C (H-2d) mice skin. In group D, C57BL/6 (H-2b) mice were treated with HSP60 emulsified in IFA into the back and followed by skin transplantation of CBA/N (H-2k) mice 2 weeks later. The delayed type hypersensitivity was determined 7 days after transplantation. One-way mixed lymphocyte reaction, the concentration of cytokines in the mixed lymphocyte reaction culture supernatant was determined 7 days and 25 days after transplantation. The survival time of skin allograft was observed. Results The survival time of skin allograft in groups A, B, C and D was 12.4 ± 0.5, 11.6 ± 0.8, 29.3 ± 2.6 and 27.6 ± 2.1 days, respectively. There was significant difference between groups A, B and groups C, D (P﹤0.05), while there was no significant difference between group A and group B as well as between group C and group D (P gt; 0.05). The counts of per minute impulse (cpm) of mixed lymphocyte reaction 7 days after transplantation in groups A, B, C and D was 12 836 ± 1 357, 11 876 ±1 265, 6 581 ± 573 and 6 843 ± 612, respectively. There was significant difference between groups A, B and group C and group D (P lt; 0.05), while there was no significant difference between group A and group B as well as between group C and group D (P gt; 0.05). The cpm of mixed lymphocyte reaction at 25 days after transplantation in group A, B, C and D was 13 286 ±1 498, 12 960 ± 1 376, 11 936 ± 1 265 and 12 374 ± 1269, respectively. There was no significant difference among 4 groups (P gt;0.05).The concentration of IL-10 in the mixed lymphocyte reaction culture supernatant in groups C, D were higher than that in groups A, B, and IL-2 and IFN-γ were lower than that in groups A, B 7 days after transplantation (P lt; 0.05), while there was no significant difference between group A and group B as well as between group C and group D (P gt; 0.05). There was no significant difference in cytokines among the 4 groups 25 days after transplantation (P gt; 0.05). The delayed type hypersensitivity in groups A, B, C and D 7 days after transplantation was 0.84 ± 0.09, 0.81 ± 0.07, 0.43 ± 0.05 and 0.46 ± 0.03 mm, respectively. There was significant differences between groups A, B and groups C, D (P lt; 0.05). While there was no significant difference between group A and group B as well as between group C and group D (P gt; 0.05). Conclusion HSP60 may play a role in induction and maintenance of murine skin allograft tolerance.
Objective
To study the relationship between the expression ratio of heat shock protein (HSP) 70 to C-fos in organs outside the brain after brain concussion and the time of injury in rats, in order to provide a new visual angle for determining injury time of brain concussion.
Methods
The model of brain concussion was established through free falling method. Then the rats were executed at 30 minutes, 1 hour, and 3, 6, 12, 24, 48, 96, 168, 240, 336 hours after injury. Immunohistochemistry staining of C-fos and HSP70 were used in the materials from the main organs including heart, liver, spleen, lung and kidney. All related experiment results were studied by using a microscope with image analytical system and homologous statistics.
Results
From 30 minutes to 6 hours after injury, the proportion of HSP70 immuno-positive cells increased slowly, while the proportion of C-fos immuno-positive cells increased rapidly, and the ratio of HSP70/C-fos positive cells was on the decline. From 6 to 12 hours after injury, the proportion of HSP70 immuno-positive cells rose continuously, while the proportion of C-fos immuno-positive cells started to decrease, and the HSP70/C-fos ratio showed a rising tendency. From 12 to 336 hours after injury, the proportion of HSP70 immuno-positive cells decreased slowly, while the proportion of C-fos immuno-positive cells decreased rapidly, and the HSP70/C-fos ratio was still on the rise.
Conclusions
The proportion of positive cells and ratio of the two markers in the main organs including heart, liver, spleen, lung and kidney are similar to those in the brain of rats after brain concussion. Observing the proportion of positive cells of the two markers together with their ratio in the main organs outside the brain may provide a reference for the determination of injury time after brain concussion.
