Inhibitory effect of specnuezhenide on high glucose-induced human retinal microvascular endothelial cell injury and its mechanism / 中华实验眼科杂志

Objective:To investigate the effect of specnuezhenide on high glucose-induced human retinal microvascular endothelial cells (hRMECs) injury and its mechanism.Methods:The hRMECs were divided into a normal control group cultured in a culture medium containing 5.5 mmol/L glucose, a hypertonic group cultured in a culture medium containing 5.5 mmol/L glucose + 24.5 mmol/L mannitol, a high glucose group cultured in a culture medium containing 30 mmol/L glucose, as well as high glucose+ low-, medium-, and high-dose specnuezhenide groups cultured in culture media containing 30 mmol/L glucose + 25, 50, 100 μmol/L specnuezhenide for 24 hours, respectively.In addition, hRMECs were divided into a high glucose+ small interfering RNA-negative control (si-NC) group cultured in a culture medium containing 30 mmol/L glucose, a high glucose+ si-forkhead box O4 (FOXO4) group cultured in a culture medium containing 30 mmol/L glucose, a high glucose+ specnuezhenide+ pcDNA group cultured in a culture medium containing 100 μmol/L specnuezhenide + 30 mmol/L glucose, and a high glucose+ specnuezhenide+ pcDNA-FOXO4 group cultured in a culture medium containing 100 μmol/L specnuezhenide+ 30 mmol/L glucose for 24 hours after transfection by corresponding reagents.Cell apoptosis was detected by flow cytometry.The malondialdehyde (MDA) concentration and superoxide dismutase (SOD) activity in cells were detected by the thiobarbituric acid method and xanthine oxidase method, respectively.The concentrations of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in the cell culture supernatant were detected by enzyme linked immunosorbent assay.The relative expression level of FOXO4 protein in cells was determined by Western blot.Results:The apoptosis rates of normal control group, hypertonic group, high glucose group, high glucose+ low-, medium- and high-dose specnuezhenide groups were (7.32±0.72)%, (7.44±0.70)%, (23.96±1.32)%, (19.84±1.09)%, (14.13±0.85)% and (9.84±0.70)%, respectively.There were significant differences in cell apoptosis rate, MDA concentration, SOD activity, the concentration of IL-1β, the concentration of TNF-α, and the relative expression level of FOXO4 protein among the six groups ( F=498.545, 1 186.693, 516.629, 654.247, 638.238, 472.655; all at P<0.001). Compared with high glucose group, the apoptosis rate, MDA concentration, IL-1β and TNF-α concentration, FOXO4 protein expression level were significantly decreased in high glucose+ low-, medium- and high-dose specnuezhenide groups, and SOD activity was significantly increased in a dose-dependent manner.Compared with high glucose+ si-NC group, the expression level of FOXO4 protein, cell apoptosis rate, MDA concentration, IL-1β and TNF-α mass concentrations were decreased in high glucose + si-FOXO4 group, while the SOD activity was increased.Compared with high glucose+ specnuezhenide+ pcDNA group, the apoptosis rate, MDA concentration, IL-1β and TNF-α concentrations, FOXO4 protein expression level of hRMECs in high glucose+ specnuezhenide+ pcDNA-FOXO4 group were significantly increased, and SOD activity was significantly decreased (all at P<0.05). Conclusions:Specnuezhenide can protect hRMECs from high glucose-induced apoptosis, oxidative stress and inflammatory response by down-regulating FOXO4.

Epidemiological situation of hepatitis D in the gathering area of Mongolian population in Inner Mongolia Autonomous Region of China / 临床肝胆病杂志

Objective To investigate the status and molecular epidemiology of hepatitis D virus (HDV) infection in the gathering area of Mongolian population in Inner Mongolia Autonomous Region of China. Methods A total of 230 patients with positive hepatitis B surface antigen (HBsAg) who attended Inner Mongolia International Mongolian Hospital from April 2019 to October 2020 were enrolled, and according to related information, they were divided into hepatitis B+liver cirrhosis group( n =18) and hepatitis B group( n =212). According to HBsAg quantification with a cut-off value of 250 IU/mL, the patients were divided into HBsAg < 250 IU/mL group( n =104) and HBsAg ≥250 IU/mL group( n =126). ELISA was used to detect HDV antibody, and quantitative real-time PCR was used to measure HDV RNA in patients with positive HDV antibody. Genotyping was performed for HDV RNA-positive samples. The chi-square test was used for comparison of categorical data between two groups. Results The positive rate of HDV antibody was 16.09%, and among the patients with positive HDV antibody, the positive rate of HDV RNA was 91.89%. Among the 18 patients with hepatitis B and liver cirrhosis, the positive rate of HDV antibody was 44.44%, and among the patients with positive HDV antibody, the positive rate of HDV RNA was 100%. There were 104 patients with HBsAg < 250 IU/mL, among whom only 3 patients (2.88%) were positive for hepatitis D antibody, and there were 126 patients with HBsAg ≥250 IU/mL, with a positive rate of HDV antibody of 26.98%. Genotype 1 was observed in all the samples that could be genotyped. Conclusion There is a relatively high infection rate of HDV in Inner Mongolia Autonomous Region, especially in patients with HBsAg ≥250 IU/mL or those with liver cirrhosis. It is necessary to strengthen the detection of hepatitis D in HBsAg-positive patients and perform early diagnosis and treatment to prevent the further progression of hepatitis.

Effects of Citicoline Acupoint Injection on Expression of Growth Associated Protein-43 in Rats with Traumatic Brain Injury / 中国康复理论与实践

@#Objective To investigate the effects of injection of citicoline into Zusanli (ST36) acupoint on neural function and expression of growth associated protein-43 (GAP-43) in rats with traumatic brain injury. Methods 40 healthy adult male Sprague-Dawley rats were randomly divided into 5 groups: sham-operated group (group A, n=8), acupoint injection of citicoline group (group B, n=8), acupoint injection of saline group (group C, n=8), intraperitoneal injection of citicoline group (group D, n=8) and intraperitoneal injection of saline group (group E, n=8). Opened brain trauma was induced with the modified Feeney method in the groups B, C, D and E, and were treated as design, once a day for 14 days. They were assessed with nervous function score and open-field test before and 8, 14, 15, and 22 days after injury. The expression of GAP-43 in the brain were detected with immunohistochemistry 28 days after injury. Results The nervous function scores and open-field test scores improved more significantly in the group B than in the groups C, D and E (P<0.05). The expression of GAP-43 increased in the group B than in the groups C, D and E (P<0.05). Conclusion Acupoint injection of citicoline into Zusanli may improve the expressions of GAP-43 to promote the recovery of neural function in rats after traumatic brain injury.

Effects of Citicoline Acupoint Injection on Expression of Growth Associated Protein-43 in Rats with Traumatic Brain Injury / 中国康复理论与实践

Objective To investigate the effects of injection of citicoline into Zusanli (ST36) acupoint on neural function and expression of growth associated protein-43 (GAP-43) in rats with traumatic brain injury. Methods 40 healthy adult male Sprague-Dawley rats were ran-domly divided into 5 groups:sham-operated group (group A, n=8), acupoint injection of citicoline group (group B, n=8), acupoint injection of saline group (group C, n=8), intraperitoneal injection of citicoline group (group D, n=8) and intraperitoneal injection of saline group (group E, n=8). Opened brain trauma was induced with the modified Feeney method in the groups B, C, D and E, and were treated as de-sign, once a day for 14 days. They were assessed with nervous function score and open-field test before and 8, 14, 15, and 22 days after inju-ry. The expression of GAP-43 in the brain were detected with immunohistochemistry 28 days after injury. Results The nervous function scores and open-field test scores improved more significantly in the group B than in the groups C, D and E (P<0.05). The expression of GAP-43 increased in the group B than in the groups C, D and E (P<0.05). Conclusion Acupoint injection of citicoline into Zusanli may im-prove the expressions of GAP-43 to promote the recovery of neural function in rats after traumatic brain injury.

New development in the treatment of sepsis / 医学研究生学报

Sepsis constitutes a serious threat to human health and represents a major burden to the health care system.Recent years have witnessed a deeper insight into the mechanism of the onset of sepsis,together with some new therapeutic strategies,including early aggressive volume therapy,activated protein C,potentiated insulin therapy,adrenal function replacement therapy,and some new therapeutic targets,which have offered some hope for the management of sepsis.

Effects of different crystalloid fluids at different osmotic concentrations on blood-brain barrier and cerebral edema in hemorrhagic shock rats / 医学研究生学报

Objective:To compare the effects of 3 different crystalloid fluids at different osmotic concentrations on blood-brain barrier(BBB) and brain edema in severe hemorrhagic shock rats.Methods: A total of 150 male SD rats were equally randomized into a lactated Ringers(LR) group,a 7.2% hypertonic saline(HS) group and a plasmalyte A(PA) group.LR,PA and HS were administered after an hour of severe hemorrhagic shock induced by drawing out about 40% of total blood and maintaining MAP at 35-45 mmHg.Serum S100B,cerebra1 Evans Blue(EB) and water content were determined before(T_0) and 1 h after bleeding(T_1) and immediately(T_2),1 h(T_3) and 2 h(T_4) after administration.The changes of BBB in the hippocampus CA1 area were observed by electron microscopy.Results: The serum S100B level was obviously higher at T_1,T_2,T_3 and T_4than at T_0 in all groups(P0.05).The cerebra1 water content was significantly increased at T_1,T_2,T_3 and T_4in the LR group,at T_1in the HS and at T_1,T_2 and T_3 in the PA as compared with T_0(P

Preparation and study on the physicochemical characteristics of glycoprotein from Crassostrea gigas / 中国海洋药物

Objective To study the preparation and the physicochemical characteristics of glycoprotein from Crassostrea gigas.Methods Oyster collected in Qingdao was taken as the experiment materials,and was extracted with H2O in low temperature.The extracts were purified using Sephacryl s-100 HR gel chromatography and HPLC.Three glycoprotein F22,F33,F42 were finally isolated from the oyster extracts.Results The physicochemical characteristics of glycoprotein were studied.The results showed that the pI value of F22 was 5.5,relative molecular mass of F22 was 34230Da,the protein contents of F22,F33,F42 were 34.1%,19.7% and 12.5%,the saccharide contents of F22,F33,F42 were 35.9%,28.8% and 40%.Conclusion The research result would be used for the study of bioactive component from Crassostrea gigas.

Study of isolation,preparation and structure identification of peptides from hydrolyzates of oyster / 中国海洋药物

Objective To study the preparation and the structure identification of peptides from hydrolyzates of oyster. Methods The oyster protein was hydrolyzed with trypsin. The hydrolysates were purified using Sephadex G25 gel chromatography,DEAE-Sepharose FF ion exchange chromatography,reversed-phase HPLC on C18 column. Results The peptide F32 was finally isolated from the oyster hydrolysates.The oyster protein was hydrolyzed with trypsin. IR spectrum showed the conformation of peptide chain was ?-helix.the amino acid sequence of F32 was determined by ESI-MS/MS. The sequence of F32 was as follows:Arg-Gln-Ile or Leu-Gly-Ala-Thr-Asn-Ala. Conclusion The research result would provide foundation for the structure-activity research of the peptide F32.

EXPERIMENTAL RESEARCH WITH PCR AND DNA HYBRIDIZATION TECHNIQUE IN DETECTION AND DIAGNOSIS OF ECHINOCOCCUS GRANULOSUS / 中国血吸虫病防治杂志

Objective To explore the application of PCR in detection and diagnosis of Echinococcus granulosus and develop a diagnostic technique by DNA hybridization labeled with DIG.Methods A pair of primers were designed according to Echinococcus granulosus gene fragment sequence, and the special primers were as: P1 5′-GGAATGGAGAGAAGTTAC-3′ P2 5′-GCAACCTCCGGAACTTGC-3′. The hydatid fluid, secondary hydatid and protoscolex of Echinococcus granulosus were used as template, 417 bp special band was got after being amplified by PCR. The PCR product was purified and labeled with DIG. The special DNA probe was successfully got and was used to detect the Echinococcus granulosus. Results The DNA from Escherichia coli, Shigella, Tubercle, Cysticercus granulosus and healthy human leukocyte were extended by PCR and hybrided by DNA probe labeled with DIG, and only Cysticercus granulosus expressed 471 bp special band. The sensitivity of PCR was that one Cysticercus granulosus can be detected or 100-10 fg DNA, while the dot hybridization was 2500 fg. Heterologous DNA did not be positive reaction, even increasing the spotting membrane dosages. Conclusion The technique of PCR labeled with DIG showed good specificity, high sensitivity, more accuracy and quickness and so it can provide scientific basis for early diagnosis and epidemiological investigation of Echinococcus granulosusEXPERIMENTAL RESEARCH WITH PCR AND DNA HYBRIDIZATION TECHNIQUE IN DETECTION AND DIAGNOSIS OF ECHINOCOCCUS GRANULOSUS$$$$ Feng Xiaomei, Ji Hugang, Cao Yinfang, Wang Wenhao (Department of Clinical Laboratory, Inner Mongolia Autonomous Region Hospital, Huhhot 010017, China) Abstract Objective To explore the application of PCR in detection and diagnosis of Echinococcus granulosus and develop a diagnostic technique by DNA hybridization labeled with DIG.Methods A pair of primers were designed according to Echinococcus granulosus gene fragment sequence, and the special primers were as: P1 5′-GGAATGGAGAGAAGTTAC-3′ P2 5′-GCAACCTCCGGAACTTGC-3′. The hydatid fluid, secondary hydatid and protoscolex of Echinococcus granulosus were used as template, 417 bp special band was got after being amplified by PCR. The PCR product was purified and labeled with DIG. The special DNA probe was successfully got and was used to detect the Echinococcus granulosus. Results The DNA from Escherichia coli, Shigella, Tubercle, Cysticercus granulosus and healthy human leukocyte were extended by PCR and hybrided by DNA probe labeled with DIG, and only Cysticercus granulosus expressed 471 bp special band. The sensitivity of PCR was that one Cysticercus granulosus can be detected or 100-10 fg DNA, while the dot hybridization was 2500 fg. Heterologous DNA did not be positive reaction, even increasing the spotting membrane dosages. Conclusion The technique of PCR labeled with DIG showed good specificity, high sensitivity, more accuracy and quickness and so it can provide scientific basis for early diagnosis and epidemiological investigation of Echinococcus granulosus[infections.