Endoplasmic reticulum stress,an critical component of cell stress,is a protective response for eukaryotic cells. The change of partial pressure of oxygen (for instance hypoxia) is a primary condition for cell stress,significantly impacting the biological phenotype and behavior of cells (including survival,migration and invasion et al). Through endoplasmic reticulum stress,intracellular concentration of unfolded protein is reduced,and agglutination inhibited. Recently,endoplasmic reticulum stress has been demonstrated to play an important role in pathogenesis of some diseases such as cancer,diabetes and inflammation,which are characterized by the micro-environment hypoxia in local tissue. Therefore,investigations on the mechanism of endoplasmic reticulum stress caused by hypoxia may promote the novel therapeutic strategy for tumors,cardiovascular diseases and diabetes.

Objective: To screen for the differentially expressed proteins during rat liver regeneration after partial hepatectomy (PH) by proteomics technique. Methods: Healthy male SD rats were randomly divided into 2 groups: hepatectomy group and sham operated group. The hepatectomy model was produced by 70% PH (n=35) and the sham operated rats (n=5) underwent the same surgical protocol without hepatectomy. Rats were executed at 2, 12, 24, 36, 48, 72 and 168 h after partial hepatectomy (each time 5 rats) and the right lobes were harvested. The total protein was extracted and analyzed by two-dimensional gel electrophoresis and mass spectrometric analysis. The differential proteins were then analyzed by Western blotting. Results: The spots of differential protein began to rise at 2 h after PH and peaked at 36 h after PH. A total of 78 protein spots were identified and 35 significant protein spots were found by mass spectrometric analysis. The 35 protein sports fell into 5 types according to their dynamic changes: 3 up-regulated and 2 down-regulated (with different regulation time periods and amplitudes); and their functions involved oxidative stress response, acute reaction, lipid and energy metabolism, intracellular signaling transduction, cell proliferation, etc., with some having unknown functions. Western blotting analysis showed that the prohibitin protein began to increase 2 h after PH and decreased to the normal levei after 48 h. Conclusion: It is indicated that many proteins and signal transduction pathways participate in the liver regeneration after partial hepatectomy.

AIM: To establish the quality control for Lidanzhitong Tablet (Radix Paeoniae Rubra,Fructus Aurantii,Rhizoma Corydalis,etc.). METHODS:Radix Paeoniae Rubra, Fructus Aurantii and Rhizoma Corydalis were identified by TLC, and the content of paeoniflorin in Radix Paeoniae Rubra was determined by HPLC. RESULTS: Paeoniflorin showed a good linear relationship at a range of 0.597-4.997 ?g, r=0.999 9. The average recovery was 99.06%, and RSD was 3.4%. CONCLUSION: The method is accurate and reliable, and can be used to control the quality of this preparation effectively。

Objective To study the effect of Chinese medicine combined with large dose interferon in the postoperative adjuvant therapy of malignant melanoma of the skin.Methods A high dose of interferon alpha-2b (GanLeneng) (1200 IU/per day) was used in 20 cases of malignant melanoma of skin for 7 days.One course of treatment was 3 months,8 consecutive courses of treatment and traditional Chinese medicine (0.25 g) were administered 1 hour before meals,2 times a day,taking three weeks per month.Results One patient in 20 cases of patients did not receive timely treatment and resulted in death.The remaining patients were were still live from 2012 to the present.Local and distant metastasis lesions disappeared during the course of treatment.Conclusions Traditional Chinese medicine combined with high-dose interferon in cutaneous malignant melanoma as adjuvant treatment plays an important role.

BackgroundHypoxia and hyperglycemia are the common causes of retinal neovascularization.In these states,H+ accumulates because of the elevated glycolysis and failure of retinal circulation,thus the retinas readily acidified. ObjectiveThe present study was to explore whether retinal acidosis independently regulates the production of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) and whether the regulation is related to oxidative stress.Methods The retinas from 2-week-old male SD rats were cultured with explant method in DMEM modulated by NaHCO3,and culture retinas were randomly divided into pH 7.2,6.8 and 6.5 groups for 24 hours.In addition,after 24 hours of culture as above described,retinas were washed using PBS two times and then followed by again culture in DMEM with pH 7.2 for another 24 hours.Also,antioxidant was added in different pH values of DMEM for culture as above described.The retinal samples were prepared for histopathological examination.The expressions of VEGF and PEDF proteins and their mRNA in retina tissue were detected by Western blot and fluorescence quantitative polymerase chain reaction (PCR) respectively.Results The retina showed the clear structure and morphology in pH 7.2 group and pH 6.8 group,but retinal vacuoles change was seen in pH 6.5 group after culture for 24 hours.No significant difference was seen in the expressing level of VEGF mRNA in retina between normal group and pH 7.2 group( 112％ ±11％ vs 100％ ±7％ ) (P=0.55),but those in pH 6.8 group and pH 6.5 group were significant increased in comparison with pH 7.2 group( 196％ ±43％ vs 100％ ±7％ ;251％ ±29％vs 100％ ±7％ )( P＜0.05 ).The expressing level of PEDF mRNA in retina in normal group was similar to that of pH7.2 group(86％ ±19％ vs 100％ ±33％) (P=0.64),but that in pH 6.5 group was significantly higher than pH 7.2 group( 230％ ±66％ vs 100％ ±33％ ) ( P＜0.05 ).The resemble results were found in the expressions of VEGF and PEDF protein.After pH reversion,the expressing levels of VEGF mRNA were 100％ ±13％,111％ ±9％,113％ ±9％ in pH 7.2 group,pH 6.8 group and pH 6.5 group respectively without significant difference among them (F=2.51,P=0.16).The expressing levels of PEDF mRNA were 100％ ±13％,110％ ±9％,108％ ±11％in different groups ( F =0.98,P =0.43 ).Under the presence of antioxidant,the expressing level of VEGF mRNA in pH 6.5 group increased in comparison with pH 7.2 group and pH 6.8 group ( P ＜ 0.05 ).The expressing levels of PEDF mRNA were significant different among pH 7.2 group( 100±31 )％,pH 6.8 group( 282±45 )％ and pH 6.5 group(480±117)％ (F=20.73,P=0.00). Conclusions VEGF can be induced by retinal acidification alone,which may be regulated by oxidative stress.Under the retinal acidification,antioxidants promote the expression of PEDF,suggesting that oxidative stress inhibits the production of PEDF.

Objective:To investigate the clinical efficacy of microscopic myringoplasty versus endoscopic myringoplasty in the treatment of tympanic membrane perforation caused by chronic suppurative otitis media. Methods:The clinical data of 91 patients with tympanic membrane perforation caused by chronic suppurative otitis media who received treatment in Jiaxing Second Hospital, China between February 2017 and March 2019 were retrospectively analyzed. These patients were divided into a control group ( n = 45) and an observation group ( n = 46) according to different surgery methods. The control group was given microscopic tympanoplasty, while the observation group was given endoscopic tympanoplasty under the otoendoscope. Results:Blood loss in the observation group was significantly lower than that in the control group [(7.2 ± 2.0) mL vs. (13.7 ± 3.1) mL, t = 11.912, P < 0.001]. Operation time in the observation group was significantly shorter than that in the control group [(59.4 ± 5.4) min vs. (91.5 ± 11.2) min, t = 17.474, P < 0.001]. Postoperative pain score in the observation group was significantly lower than that in the control group [(2.9 ± 0.7) points vs. (4.8 ± 1.3) points, t = 8.707, P < 0.001]. Hospital stay in the observation group was significantly shorter than that in the control group [(4.3 ± 1.0) d vs. (6.5 ± 1.5) d, t = 8.249, P < 0.001]. Pure tone hearing thresholds at 1, 2 and 4 kHz frequencies in the observation group were significantly higher than those in the control group (all P < 0.05). Patient satisfaction regarding the aesthetic effect of the surgical incision in the observation group was significantly higher than that in the control group [97.8% (45/46) vs. 77.8% (35/45), χ2 = 8.604, P = 0.003]. Conclusion:Endoscopic myringoplasty has the advantages including shorter operation time, less blood loss, lower degree of pain, better hearing improvement and higher patient satisfaction over microscopic myringoplasty in the treatment of tympanic membrane perforation caused by chronic suppurative otitis media.

ObjectiveTo observe the effect of rehabilitation on fractures around knee treated with external fixation.Methods48 cases with the fractures around knee who accepted early rehabilitation after external fixation had been observed for 0.5～4 years.ResultsAll the cases came up to clinical healing standards within 7 months. According to Kolmert's standard, the rate of fine and good was 89.6%.ConclusionThere was satisfactory outcome treating fractures around knee with external fixation. Early rehabilitation is important to avoid the knee joint rigor.

Objective: To establish the quality standard for Heluoshugan Capsules(Rhizoma Atractylodis macrocephalae, Radix Paeoniae alba, Rhizoma cyperi, Rhizoma sparganii, Radix et Rhizoma rhei, Radix Angelicae Sinensis, etc.). Methods: Paeoniflorin of Radix Paeoniae Alba was determined by HPLC. Rhizoma cyperi, Radix Paeoniae alba, Radix aucklandiae and Rhizoma sparganii were identified by microscopic identification, and Radix et Rhizoma Rehei was identified by TLC. Results: The average recovery rate of paeoniflorin was 99.18%, and RSD was 2.3% . The microscopic characteristics were obvious. The TLC sports developed were fairly clear, and the blank test showed no interference. Conclusion: The method developed is accurate and the reproducibility is good. The method can be used for quality control of the Heluoshugan C apsules.

Objective To evaluate the influence of narrow-band ultraviolet B on lesional microvessel density (MVD),vascular endothelial growth factor (VEGF),matrix metalloproteinases 2 (MMP-2)as well as on serum VEGF in patients with psoriasis vulgaris(PV).Methods Fifteen patients with PV were recruited into this study with 10 normal human controls.All patients received NB-UVB phototherapy thrice a week for 4-5 weeks.Prior and after the treatment,psoriasis area and severity index (PASI)was calculated,tissue specimens were taken from non-photoexposed lesions,and sera samples were obtained from these patients.Then,MvD and the expression level of VEGF and MMP-2 were measured by immunohistochemical labeled dextran polymer(LDP)method in the tissue specimens.Also,the serum level of VEGF was tested by enzyme-linked immunosorbent assay(ELISA).Results PASI score remarkably decreased in patients after the photothempy(t=13.35,P＜0.01).The MVDs were 20.52±5.02,7.33±1.24 and 4.26±0.79 capillaries per high power field(400 × amplification),in psoriatic lesions before treatment,after treatment,and normal control tissues,respectively,with a significant difference among the three groups (F=97.57,P＜0.05),and a significant increase was observed in the lesions before treatment compared with those after treatment and normal controls.The serum level of VEGF was 307.55±121.65 ng/L in psoriatic lesions before treatment,significantly higher than that after treatment(163.92±95.57 ng/L),and in normal control skin (139.78±79.06 ng/L),whereas there was no significant difference between the latter two groups(P＞0.05).The positivity rate of MMP-2 was similar among the three groups without statistical difference(P＞0.05).In psoriatic patients,a positive correlation was observed among PASI score,MVD,lesional and serum VEGF levels(P＜0.05),also among the MVD,VEGF and MMP-2 levels in lesions(P＜0.05).but lesional MMP-2 was unrelated to PASI score or sgrum VEGF(both P＞0.05).Conclusions NB-UVB may regulate superficial dermal microvascular proliferation by acting on the expression of VEGF in sera and lesions of psoriatic patients.VEGF and MMP-2 may bOth participate in the proliferation process of microvessels,while MMP-2 is unlikely to be involved in the therapeutic mechanism of NB-UVB.

Objective To investigate the expression of DNA methyltransferases ( DNMTs) in hilar cholangiocarcinoma and its clinical significance.Methods A total of 150 samples of cholangetic tissues were collected from 111 patients with hilar cholangiocarcinoma ( cholangiocarcinoma group) and 39 patients with choledochocele ( control group) at the First Affiliated Hospital of Sun Yat-Sen University from April 1997 to March 2007.A tissue chip containing the samples of hilar cholangiocarcinoma and choledochocele was prepared.Expressions of DNMT1,DNMT3a and DNMT3b were detected by the immunohistochemical staining. Differences in the protein expressions of DNMTs in the cholangiocarcinoma group and the control group were compared,and the correlation between DNMTs protein expressions and clinicopathological features was analyzed.All data were analyzed by using the chi-square test or Fisher exact probability.The survival curve was drawn by using the Kaplan-Meier method and the survival rate was compared by using the Log-rank test.Results The rates of high protein expressions of DNMT1 and DNMT3b were 54.1％ (60/111) and 47.7％ (53/111) in the cholangiocarcinoma group, which were significantly higher than 28.2％ ( 11/39) and 23.1％ ( 9/39) in the control group　( x2 =7.740,7.240,P ＜0.05). The high protein expression of DNMT1 was correlated with-the Bismuth-Corlette classification and T staging of the tumor ( x2 =12.200, 17.800,P ＜0.05) ; there was no significant difference in the high protein expressions of DNMT3a in the cholangiocarcinoma group and the control group ( x2 =3.370.P ＞0.05 ) ; while the high protein expressions of DNMT3b was correlated with the Bismuth-Corlette classification (x2 =8.300,P ＜ 0.05 ),but not with the T staging. Sixty-six patients received hilar cholangiocarcinoma resection,and 42 of them were followed up.The median postoperative survival time of patients with low protein expression of DNMT1 was 23.9 months,which was significantly longer than 11.8 months of patients with high protein expression of DNMT1 (x2 =3.980,P ＜ 0.05).Conclusions DNMT1 and DNMT3b with high protein expression might play important roles in the carcinogenesis and development of hilar cholangiocarcinoma.There is an obvious relationship between the expression of DNMT1 and postoperative survival time of patients with hilar cholangiocarcinoma,and DNMT1 might be a valuable prognostic factor for hilar cholangiocarcinoma.