Objective To investigate the effects of sericin pretreatment on the expression of extracellular matrix(ECM) associated protein in diabetic nephropathy(DN) rats' kidney.Methods Sixty six male SD rats were randomly divided into 3 groups(n=12):normal control group,DN model group and sericine pretreatment group.DN rats model in model group and sericine pretreatment group were established by intraperitoneally injection of streptozotocin(STZ).Blood glucose≥16.7 mmol/L was taken as the standard of successful modelization.The rats in sericine pretreatment group were lavaged with sericine(2.4 g·kg~(-1)·d~(-1)) for 35 days before injecting STZ.The enzymic method was used to measure the blood glucose.Type Ⅳ collagen(cⅣ)and laminin(LN)content in the serum were detected by ELBA.The expression of transforming growth factor-β_1,(TGF-β_1)and tissue inhibitors of maprix metalloproteinase-1(TMP-1) protein in the kidney was observed by immunohistochemical staining.The expression of Smad 3 protein in the kidney was detected by Western blot.Results Compared with normal control rats,the blood glucose,cⅣ and LN content in the serum,TGF-β_1,TIMP-1 and Smad 3 expression in the kidney of the model group rats increased obviously(P<0.01).The blood glucose,cⅣ and LN content in the serum,TGF-β_1,TMP-1 and Smad3 expression in the kidney of rats in sericine pretreatment group were significantly lower than those of the rats in model group(P<0.01).Conclusion Sericin pretreatment can inhibit the activation of TGF-β/Smad 3 signal pathway in the kidney of DN rats,and prevent the decrease of MMPs activity induced by up-regulation of TIMP-1.So sericin can prevent accumulation of ECM and glomerulosclerosis during DN,and has satisfactory apotropaic effects on the development of DN.

Objective To investigate the effects of hydrocortisone (HC) on proliferation and killing activity of NK cells against pancreatic cancer SW1990 cells in vitro.Methods Peripheral blood mononuclear cells of healthy people were isolated and cultured with NK cells medium containing IL-1S.When the purity of NK cells reached above 70％,different concentrations of HC (10-6,10-5,10-4,10-3 μmol/L) were added and co-cultured with NK cells for 7 days.And NK cells without HC were used as control.CD3-CD56 + NK cell numbers of each group were countered by trypan blue staining.Perforin,granzyme B and IFN-γ expression of CD3-CD56+ + NK cells were verified by flow cytometry.NK cells and SW1990 cells were co-cultured with a 20∶1effector to target ratio,then the cytotoxic activity of NK cells against SW1990 cells were analyzed by CCK-8 kit.Results After treatment with different concentration of HC for 7 days,NK cells purity of each group reached 70.72％ ～ 76.39％,and it was not significantly different with that in control group [(72.61 ± 3.76) ％].The proliferation folds of NK cells treated with 10-6,10-5,10-4,10-3 μmol/L HC were (9.13 ± 0.94),(9.67 ±1.51),(10.33±1.07),(8.40±1.47) times,respectively,while it was (4.23 ±0.82) times in control group (all P ＜0.01).The killing effects of NK cells on SW1990 cells were (58.58 ± 4.89) ％,(62.27 ± 5.63) ％,(64.02 ± 5.79) ％,(63.88 ± 3.61) ％,which were higher than that in control group [(57.46 ± 5.11) ％],moreover,the difference between NK cells of 10-4 μmol/L HC treatment group and control group was statistically significant(P ＜ 0.05).The expressions of perforins of 10-4,10-3 μmol/L HC treatment group were (96.71 ± 3.04) ％,(97.56 ± 2.18) ％,which were significantly higher than that in control group [(92.40 ±3.53)％,P ＜0.05 or 0.01].The expression of granzyme B in 10-5 μmol/L HC treatment group was (78.23 ±2.94)％,which were significantly higher than that in control group [(73.68 ±3.52) ％,P ＜0.05].The expressions of IFN-γ in 10-5,10-4,10-3 μmol/L HC treatment group were (96.61 ±2.04)％,(97.58 ± 2.17)％,(98.00 ± 1.77)％,which were significantly higher than that in control group [(92.44 ± 2.74)％,P＜0.01].Conclusions HC can promote IL-15 activated NK cells proliferation and enhance NK cells mediated killing activity against SW1990 cells with proper concentration,and up-regulation of perforin,granzyme B and IFN-γ expression may be the main mechanisms.

To study the comparability of myocardial enzyme spectrum detection results by two different bio-chemical testing systems to provide references for mutual accredit of detection results in one lab. Following NC-CLS document EP9-A2, 8 clinical specimens were selected every day for five days and tested for AST, LDH, CK and CK-MB respectively with Siemens ADVIA1800 and Johnson & Johnson Victro350 systems. Acknowledging Siemens AD-VIA1800 as objective testing system, the results were utilized to make correlation analysis and clinical acceptability esti-mation. There was high precision in Siemens ADVIA1800 and Johnson & Johnson Victro350 (CV≤1/3CLIA′88 allowable error). It's proved by t test that the difference of the myocardial enzyme results tested with Siemens ADVIA1800 and Johnson Victro350 was statistically significant (P<0.01), while the two testing systems showed significant correlation (r>0.975). In the comparison with the Siemens ADVIA1800, the standard errors (SE%) of AST, CK and CK-MB tested with Johnson Victro350 were 4.0%-6.6%, 9.6% and 13.3%, which were accepted by clinical standard; while the SE% of LDH tested with Johnson Victro350 were 11.2%-12.7%, which were beyond clinically acceptable range. In case one item is tested by different testing systems in one laboratory, the comparison and bias estimation acceptable eval-uationare necessary to judge the clinical acceptability, which can ensure the accuracy and consistency of the results.

Objective To investigate the effect of hyperoside on proliferation and killing activity of NK cells against pancreatic cancer PANC1 cells in vitro,and explore its potential mechanism.Methods Peripheral blood mononuclear cells of healthy donors were isolated,NK cells were induced with medium contained with IL-2 and different concentrations of hyperoside (0.3,1.6,8,40 and 200 μg/ml) for 12 days.Cell viability was observed by trypan blue staining.Phenotype and perforin,granzyme B expression of NK cells were detected by flow cytometry.Killing activity of NK cells against PANC1 cells were analyzed with lactate dehydrogenase (LDH) releasing method.Results The proportion of NK cells in control group and experimental group treated with different concentration of hyperoside both reached about 80％,respectively.The proliferation of CDs-CD56 + NK cells treated by hyperoside at 0.3,1.6 and 8 μg/ml was (93.76 ±8.77),(106.67 ± 12.35) and (118.50 ± 11.51) times,respectively,which were significantly higher than (73.70 ± 9.43) times of the control group.The expressions of perforin in NK cells treated with hyperoside at 1.6,8 and 40 μg/ml were significantly higher than those of the control group [(82.34 ± 2.90) ％,(89.15 ±3.54) ％,(81.78 ± 2.81)％ vs (72.93 ± 2.06)％].The expressions of granzyme B in NK cells treated with hyperoside at 1.6 and 8 μg/ml were significantly higher than those of the control group [(87.30 ± 1.70) ％,(92.16 ±3.05)％ vs (82.35 ±2.73)％].The killing activity of NK cells against PANC1 cells treated by hyperoside at 1.6 and 8 μg/ml was significantly higher than those of the control group [(63.18 ± 3.77)％,(65.34 ± 4.97) ％ vs (52.16 ± 5.48) ％].The differences were statistically significant (all P ＜ 0.05).Conclusions Hyperoside could promote the proliferation of NK cells at certain concentrations and maybe enhance the killing effect against pancreatic cancer PANC1 cells through up-regulating the expression of perforin and granzyme B in NK cells.

To investigate the effect of long-term indwelling gastric tube on the prevention and treatment of esophageal stenosis after endoscopic submucosal dissection (ESD) for esophageal circumferential superficial cancer, data of patients with esophageal circumferential superficial cancer who underwent ESD in the First Affiliated Hospital of Nanjing Medical University from January 2018 to December 2021 were retrospectively analyzed. There were 15 patients with gastric tube placement (GTP) after ESD (the GTP group ), and 23 patients without GTP (the non-GTP group). The general information, lesion location, pathological stage, postoperative complications, degree of esophageal stenosis (water intake), pain conditions, number of hospitalizations and medical expenses were compared between the two groups. The results showed that there was no significant difference in age, gender, lesion location or postoperative pathological stage between the two groups ( P>0.05). Compared with the non-GTP group, the rate of water intake in the GTP group was significantly higher (11/15 VS 6/23, P<0.05), the frequency of pain was less in the GTP group (7.3±3.1 times VS 10.7±3.6 times, t=3.00, P<0.05), and the number of hospitalizations and the medical expenses after ESD to before and after stent placement were significantly lower in the GTP group than those in the non-GTP group ( P<0.05). There were no significant differences in the incidence of delayed bleeding and perforation, or time of the first stenosis after ESD between the two groups ( P>0.05). The results of the study initially showed that long-term indwelling gastric tube after ESD can reduce the degree of esophageal stenosis with good safety for esophageal circumferential superficial lesions.

From the perspective of state differentiation and treatment， it is believed that the pathogenesis of acute respiratory distress syndrome （ARDS） is that evil poisons injured the lungs， and the lung qi suddenly collapsed， then blocked and exhausted， and the qi failure to control blood and liquid， then the fluids overflow outside the vessels， and damp phlegm， stasis， and toxins became knotted up in the body， which ultimately leads to qi dysfunction， and a series of symptom arise， so qi impairment is the principal mechanism of ARDS. A combination of Chinese and Western medicine was proposed to treat ARDS by combining tangible qi and intangible qi， using Chinese herbal medicine to boost qi and relieve collapse， percolate and drain dampness with bland medicinals， resolve toxins and dissolve stasis， and regulate qi， and combining with Western medicine to assist qi circulation to improve qi's consolidation， propulsion， and transformation， so as to make the evil qi go away， the positive qi restored， the viscera qi circulated， qi， blood， yin， and yang connected， and the activities of life maintained， and thus to achieve the goal of treating ARDS by integrated Chinese medicine and Western medicine.

Glypican-3 (GPC3) is a key member of Glypican family and plays an important role in the development, angiogenesis and metastasis of hepatocellular carcinoma (HCC). Most HCC overexpresses GPC3, but GPC3 is hardly detected in normal adult liver and benign liver lesions, so it is regarded as a highly specific diagnostic marker and an ideal therapeutic target for HCC. In this study, we cloned the heavy and light chain variable region gene from the monoclonal antibody targeted to GPC3 screened in the previous stage, and linked it with a segment of flexible peptide (Linker) to obtain the single chain antibody against GPC3. The single chain antibody gene was cloned into vector for prokaryotic expression and purified to obtain high purity protein. Detection shows that the single-chain antibody produced by us has the same binding activity with the full-length antibody, and can accurately target the tumor site of Huh7 tumor-bearing model mice after coupling Cy5.5 fluorescence, suggesting that the single-chain antibody has the potential to realize multi-directional liver cancer precise surgical navigation under the guidance of a probe.
Animals ; Antibodies, Monoclonal ; Carcinoma, Hepatocellular/genetics* ; Glypicans/genetics* ; Liver Neoplasms/diagnosis* ; Mice