AIM: To investigate the effects of salidroside on intracellular free calcium concentration (([Ca~(2+)]i)), apoptosis, mitochondrial membrane potential (MMP) and activity during injury induced by hypoxia/hypoglycemia in cultured SH-SY5Y cells. METHODS: Mitochondrial activity was measured by methylthiazolyl tetrazolium test. MMP, [Ca~(2+)]i and apoptosis were measured by flow cytometry. RESULTS: SH-SY5Y cells were cultured in a hypoxia/hypoglycemia condition for 2, 4, 6 and 12 h, [Ca~(2+)]i and apoptosis rate significantly increased compared with control group (P

Objective To investigate the effects of protocadherin 20 (PCDH20) in the spinal cord in the development of bone cancer pain (BCP) in rats.Methods Thirty-six SPF female Sprague-Dawley rats,weighing 180-200 g,were randomly divided into 4 groups (n =9 each):sham operation group (group S),BCP group,lentivirus control group (group LC) and PCDH20 siRNA lentivirus group (group P).Control lentivirus and lentivirus containing PCDH20 siRNA 4 μl were injected into the ipsilateral spinal cord in groups LC and P,respectively.One week later,BCP was induced by injection of Walker 256 breast cancer cells into the upper segment of bone marrow of right tibia.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before injection of lentivirus (T1),1 day before BCP (T2),and 7,14 and 21 days after BCP (T3-5).Three rats in each group were sacrificed after measurement of the MWT at 21 day after BCP and the tibia on the operated side was obtained for examination of invasion of the cancer cells with light microscope.The spinal cord was removed for determination of the expression of PCDH20 and postsynaptic density 95 (PSD95) protein (by Western blot) and mRNA (by RTPCR).Results In groups BCP,LC and P,the cancer cells grew out of the bone and destroyed the cortical bone seriously.Compared with group S,the MWT was significantly decreased at T3-5 in groups BCP,LC and P,the expression of PCDH20 and PSD95 protein and mRNA was up-regulated in groups BCP and LC,and the expression of PCDH20 was up-regulated in group P (P ＜ 0.05).Compared with BCP group,no significant change was found in the MWT and expression of PCDH20 and PSD95 protein and mRNA in group LC (P ＞ 0.05),and the MWT was significantly increased at T4,5 and the expression of PCDH20 and PSD95 protein and mRNA was down-regulated in group P (P ＜ 0.05).Conclusion PCDH20 is involved in the development of BCP through regulating the expression of PSD95 in the spinal cord and adjusting the function of excitatory synapse in rats.

Shaanxi Province located at Midwest inland of China was a typical iodine deficient disorders region. To investigate iodine and selenium levels of neonates in the Shaanxi sub-clinical cretinism region of China after supplement of iodine salt for nearly twenty years. We collected 56 umbilical cord blood samples from cretinous regions of Yijun County (a selenium deficient region) north of Shaanxi Province and Ziyang County (a selenium-enriched region) south of the province and from Lintong in Xi'an (a non-cretinous region for control). Among these samples 17 were collected from Ziyang, 20 from Lintong and 19 from Yijun. Seven trace elements of iodine, selenium, zinc, copper, iron, calcium and magnesium in the umbilical cord blood samples were measured and the results were processed statistically. There were no significant differences in the levels of iodine among all three counties. However, the level of selenium in Ziyang was the highest and in Yijun it was the lowest. The other trace elements such as Cu Zn Fe and Mg showed no significant difference among the three counties except for the Ca level which was lower in Yijun.The regression equation was established with the backward method of multiple regression was: Se=0.180+0.00006654 Fe-0.006 Cu-0.005956 Mg+0.1.

OBJECTIVESTo construct a hepatoma directed gene delivery system which could transfer preS2 antisense RNA to liver cancer cells specifically, and to explore a new therapeutic strategy for hepatocellular carcinoma by blocking hepatitis B virus (HBV) with antisense RNA targeting hepatocellular carcinoma.

METHODSGE7 and HA20 were synthesized and mixed with pEBAF-as-preS2, a hepatocarcinoma specific HBV antisense expression vector, to construct a novel HBV antisense RNA delivery system named AFP-enhancing 4-element complex. Nude mice bearing hepatocelluar carcinoma cells HepG2.2.15 were injected with AFP-enhancing 4-element complex via a tail vein. Total RNA from tissues was extracted, and reversal transcription-ploymerase chain reaction (RT-PCR) was used to detect the expression of preS2. Different doses of AFP-enhancing 4-element complex was injected into nude mice at different time points, and tumor diameter was measured.

RESULTSAFP-enhancing 4-element complex was constructed successfully. RT-PCR showed preS2 antisense RNA delivered by AFP-enhancing 4-element complex only expressed in liver tumor HepG2.2.15 cells of the mice. After the treatment of AFP-enhancing 4-element complex with dose of 0.2 micro g per mouse (once a week for 4 weeks), the mean tumor diameter of nude mice was significantly shorter than that of the control groups (0.995 +/- 0.35 cm vs 2.125 +/- 0.25 cm, P < 0.01).

CONCLUSIONSAn HBV antisense RNA gene delivery system targeting hepatocellular carcinoma, AFP-enhancing 4-element complex, was constructed successfully. PreS2 antisense RNA expressed specifically in hepatocelluar carcinoma cells significantly inhibits tumor growth of mice bearing hepatocarcinoma HepG2.2.15 and may have therapeutic potential in HBV related hepatocarcinoma.

Animals ; Drug Delivery Systems ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Hepatitis B Surface Antigens ; therapeutic use ; Hepatitis B virus ; genetics ; Liver Neoplasms, Experimental ; pathology ; therapy ; Male ; Mice ; Mice, Nude ; Protein Precursors ; therapeutic use ; RNA, Antisense ; therapeutic use

Objective To investigate the proportion of different types, distribution of genders, ages as well as the relative factors in inpatient with glaucoma. Design Retrospective case series. Participants 5058 cases of inpatients in Xingtai Eye Hospital, Hebei province from June 2004 to May 2009 were included. Methods Statistical analysis was conducted for 5058 cases of inpatients with glausoma. Main outcome Measures The type of glaucoma, age, gender and their percentages. Results In all 5058 cases, the patients with primary glaucoma, secondary glaucoma and congenital glaucoma accounted for 59.07%, 37.92% and 3.01% respectively. Primary angle-closure glaucoma (PACG) accounted for 88.65% in primary glaucoma, and primary open angle glaucoma (POAG) accounted for 11.35%. In PACG, acute PACG accounted for 53.15%, chronic PACG 46.85%;The female over forty accounted for 69.54%, male 26.95%. In POAG, the female over forty accounted for 69.54%, male 28.02%. From June 2004 to May 2005, POAG accounted for 11.32% in primary glaucoma, 12.44% from June 2008 to May 2009. There was no statistically significant difference. Conclusion In the central part of China, the majority of inpatients with glaucoma was PACG. It may relate to the regional,economic and cultural conditions.

Objective To screen stable express A30P a-synuclein PC12 cell as the research object,and to preliminarily investigate the activity of different glycogen systhesis kinase 3β (GSK3β) for the regulation of autophagy pathway causes a-synuclein autophagy level change and its effects on cell growth.Methods The vector of A30P a-synuclein was used to transfect to PC12 cells.G418 was used to screen stable expressed cells.Liposome method was used to transfect GSK3β-expressed plasmid,GSK3β silence plasmid,and blank plasmid into these cell lines,and stable expressed activity of GSK3β cell lines were screened.Western blot was used to detect the expression of GSK3 β after transfection and verify the transfection efficiency.Western blot was used to test groups of cells in LC-3 Ⅱ,Beclin1,and a-synuclein expressions.Methyl thiazolyl tetrazolium (MTF) method was used to detect the change of cell proliferation.Terminal dexynucleotidyl transferase (TdT) mediated dUTP nick end labeling (TUNEL) method was used to detect apoptosis.Results (1) When GSK3β expressed group was compared to GSK3β silence group,blank plasmidgroup,and blank control group,the expressions of LC-3 Ⅱ and Beclin1 were significantly increased (P ＜ 0.05),the expression of a-synuclein was significantly reduced (P ＜ 0.05),the results of MTT and TUNEL showed that the cells were the decrease of apoptosis and increase of cell proliferation,with a statistically significant difference (P ＜ 0.05).(2) When GSK3β silence group was compared to GSK3β express group,blank plasmidgroup,and blank control group,the expressions of LC-3 Ⅱ and Beclin1 were significantly reduced (P ＜ 0.05),the expression of a-synuclein was significantly increased (P ＜ 0.05),the results of MTT and TUNEL showed that the cells were the increase of apoptosis and decrease of cell proliferation,with a statistically significant difference (P ＜0.05).Conclusions (1) Activation of GSK3β activity can improve the level of cell autophagy,enhance the ability of alpha-synuclein degradation,and promote cell survival.(2) Inhibition of GSK3β activity can reduce the level of cell autophagy,weaken the ability of alpha-synucleindegradation,and reduce cell survival.(3) Autophagy is closely related to the pathogenesis of Parkinson's disease (PD).The improvement of the level of autophagy and enhancing of the degradation of asynuclein is a new way of the future treatment of PD.

OBJECTIVETo evaluate efficacy and optimal cut-off-point through cerebral vascular hemodynamic indexes (CVHI) examination to predict stroke.

METHODSA number of 20,333 people at 35 years old and over were checked by CVHI and accumulative score was calculated according to the value of detected indexes. Risk factors of stroke were investigated simultaneously. One hundred and sixty-eight patients with stroke occurred during 4-year following up. Typical syndromes and signs stroke were used as golden standard to evaluate screening efficacy of CVHI.

RESULTSScore of optimal cut-off-point of cerebral vascular hemodynamic indexes was under 75 in ROC curve analyses. Sensitivity, specificity, accuracy, positive and negative predictive values, positive and negative likelihood ratios as well as Youden's index for predicting stroke within 4 years after examination were found to be 87.50%, 67.70%, 67.86%, 2.21%, 99.85%, 2.71, 0.18 and 0.55 respectively. Sensitivity and positive predict values for predicting cerebral vascular thrombosis were superior to predicting cerebral hemorrhage. Positive predicting value in risk exposure population was higher than that of overall population. Coefficiency of variation of cerebral vascular hemodynamic examination was 4.03%. The agreement rate of examination between two physicians was 97.62% and Kappa value was 0.94.

CONCLUSIONThe score of optimal cut-off-point of cerebral vascular hemodynamic indexes examination was 75. Both Efficacy and reliability for predicting stroke seemed to be good, especially for predicting cerebral vascular thrombosis.

Adult ; Aged ; Brain ; physiopathology ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Stroke ; diagnosis ; physiopathology

Objective:To investigate the influence of perioperative probiotics supplement on short-term clinical outcomes in gastric cancer patients receiving neoadjuvant chemotherapy combined with radical gastrectomy.Methods:The prospective randomized controlled study was conducted. The clinicopathological data of 80 patients who underwent neoadjuvant chemotherapy combined with radical gastrectomy in the Affiliated Hospital of Qingdao University from July 2020 to September 2021 were selected. Based on random number table, patients were allocated into two groups. Patients undergoing perioperative probiotics supplement were allocated into the experiment group, and patients undergoing perioperative conventional treatment were allocated into the control group, respectively. Observation indicators: (1) grouping situations of the enrolled patients; (2) intraoperative situations; (3) follow-up and postoperative situations; (4) inflammation related hematological indexes. Follow-up was conducted using telephone interview and outpatient examina-tion to detect postoperative complications and startup of adjuvant chemotherapy up to October 31,2021. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M(range), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was performed using the chi-square test or Fisher exact probability. Comparison of ordinal data was analyzed using the Mann-Whitney U test. Repeated measurement data were analyzed by the repeated ANOVA or generalized estimating equation. Results:(1) Grouping situations of the enrolled patients. A total of 80 patients were selected for eligibility. There were 51 males and 29 females, aged 64(42-80)years. Of the 80 patients, there were 40 patients in the experiment group and 40 patients in the control group, respectively. (2) Intraoperative situations. All patients in the experiment group and the control group underwent radical gastrectomy successfully. Cases with yield pathologic TNM (ypTNM) stage 0, stage Ⅰ, stage Ⅱ, stage Ⅲ after neoadjuvant chemotherapy, cases undergoing laparoscopic radical gastrectomy or Da Vinci robotic radical gastrectomy, the operation time, the volume of intraoperative blood loss, cases undergoing digestive tract recons-truction as Billroth Ⅱ anastomosis or Roux-en-Y anastomosis were 2, 7, 15, 13, 19, 21,205(180-240)minutes, 50(30-60)mL, 6, 34 in the experiment group, versus 4, 6, 12, 16, 23, 17, 218(190-251)minutes, 50(43-60)mL, 11, 29 in the control group, showing no significant difference in the above indicators between the two groups ( U=683.00, χ2=0.80, U=668.00, 681.00, χ2=1.87, P>0.05). (3) Follow-up and postoperative situations. All the 80 patients were followed up for 1 month after surgery. Cases with postoperative infectious complications were 6 in the experiment group, versus 15 in the control group, showing a significant difference between the two groups ( χ2=5.23, P<0.05). The application of antimicrobial agent, time to postoperative first flatus, time to postoperative first defecation, time to tolerance of solid food, duration of postoperative hospital stay, time to postopera-tive startup of adjuvant chemotherapy were 3(3-6)doses, 53(49-66)hours, 72(62-82)hours, (72±18)hours, 6.0(5.5-7.0)days, 26.0(25.0-28.0)days in the experiment group, versus 6(3-10)doses, 66(60-88)hours, 94(82-112)hours, (107±23)hours, 7.0(6.4-8.3)days, 30.0(28.0-33.0)days in the control group, showing significant differences in the above indicators between the two groups ( U=471.50, 432.00, 343.50, t=-7.62, U=411.50, 319.50, P<0.05). (4) Inflam-mation related hematological indexes. ① The white blood cell counts before surgery and at postoperative day 1, 3, 5 were (5.6±1.4)×10 9/L, (9.9±3.2)×10 9/L, (7.7±2.6)×10 9/L, (6.8±1.8)×10 9/L in the experiment group, versus (6.1±1.9)×10 9/L, (12.3±2.9)×10 9/L, (9.7±3.6)×10 9/L, (7.8±2.7)×10 9/L in the control group, meeting the mauchly′s test of sphericity ( χ2=4.17, P>0.05). Results of intrasubject effect test showed that there were significant differences in the time effect, intervention effect and interaction effect of white blood cell counts between the two groups ( F=106.61, 10.45, 4.56, P<0.05). ② The neutrophilic granulocyte percentages before surgery and at postoperative day 1, 3, 5 were 55%±10%, 76%±11%, 73%±9%, 69%±9% in the experiment group, versus 56%±9%, 84%±5%, 79%±8%, 74%±9% in the control group, not meeting the mauchly′s test of sphericity ( χ2=16.63, P<0.05). Results of multi-variate test showed that there were significant differences in the time effect, intervention effect and interaction effect of neutrophilic granulocyte percentages between the two groups ( F=92.42, 11.46, 5.55, P<0.05). ③ The levels of C-reactive protein before surgery and at postoperative day 1, 3, 5 were 1.35(1.15-1.97)mg/L, 14.94(8.24-21.22)mg/L, 33.39(13.02-66.02)mg/L, 18.36(8.27-60.43)mg/L in the experiment group, versus 1.62(0.97-2.27)mg/L, 24.03(10.42-36.52)mg/L, 81.66(31.20-116.76)mg/L, 46.84(28.30-80.26)mg/L in the control group, not meeting the normal distribution. Results of generalized estimation equation test showed that there were significant differences in the time effect, intervention effect and interaction effect of levels of C-reactive protein between the two groups ( Waldχ2=145.74, 9.48, 9.90, P<0.05). ④ The levels of procalcitonin before surgery and at postoperative day 1, 3, 5 were 0.02(0.02-0.04)μg/L, 0.08(0.06-0.12)μg/L, 0.12(0.07-0.21)μg/L, 0.09(0.06-0.15)μg/L in the experiment group, versus 0.02(0.02-0.04)μg/L, 0.14(0.07-0.71)μg/L, 0.35(0.14-0.71)μg/L, 0.24(0.10-0.48)μg/L in the control group, not meeting the normal distribution. Results of generalized estimation equation test showed that there were signifi-cant differences in the time effect, intervention effect and interaction effect of levels of procalcitonin between the two groups ( Waldχ2=62.88, 14.71, 18.33, P<0.05). Conclusion:Perioperative supple-ment of probiotics can reduce the incidence of postoperative infectious com-plications and the application of antimicrobial agent, promote recovery of gastrointestinal function, reduce the level of inflammation related indexes, shorten the duration of postoperative hospital stay and the time to postoperative startup of chemotherapy in patients undergoing neoadjuvant chemotherapy combined with radical gastrectomy.