Objective Obstructive sleep apnea hypopnea syndrome (OSAHS) often causes damage to multiple systems, especially to the central nervous system, inducing cognitive dysfunction.This study aims to explore the possible correlation of the expressions of serum hypoxia-inducible factor-1α (HIF-1α) and brain-derived neurotrophic factor (BDNF) with cognitive impairment in rats under different hypoxia conditions.Methods Twenty-four Wistar rats were equally randomized into a normal control, a chronic intermittent hypoxia (CIH), and a chronic continuous hypoxia (CCH) group.The rats of the CIH group were placed in a hypoxia chamber filled with N2 and air, the oxygen concentration switched from (7±0.5)% to 21%, 1.5 minutes for each state and 4 minutes for each cycle, while those of the CCH group were placed in another hypoxia chamber with the oxygen concentration of (7±0.5)%, 8 hours a day and all for 30 days.Then we recorded the body weight of the rats, detected the expressions of serum HIF-1α and BDNF by ELISA, and observed the changes of behavior by Morris water maze test and those of the hippocampal morphological structure by HE staining.Results At 30 days after modeling, the body weight of the rats was significantly decreased in the CIH and CSH groups as compared with the normal control ([195.75±6.497] and [180.88±12.017] vs [218.63±15.287] g, P<0.05).Positioning navigation showed that the escape latency was significantly longer in the hypoxia models than in the controls (P<0.05), even longer in the CIH than in the CCH group (P<0.05).Spatial exploration test manifested a lower frequency of crossing the platform in the CIH and CCH groups than in the control ([2.63±1.45] and [3.22±1.30] vs [4.97±0.47] times, P<0.05).The expression levels of serum HIF-1α and BDNF were significantly higher in the CIH ([36.14±9.34] and [1625.34±332.44] pg/mL) and CCH ([27.27±6.88] and [1204.07±363.81] pg/mL) than in the normal control group ([14.11±4.06] and [1036.40±124.48] pg/mL) (P<0.05), even higher in the CIH than in the CCH group (P<0.05).HE staining exhibited scattered and disorderly arrangement of hippocampal neurons in the model rats, with unclear nuclear membrane, pyknosis of the nuclei, darkly stained cytoplasm, and some damaged cells.More obvious absence and vacuolization of some cells were observed in the rats of the CIH group.Conclusion Chronic hypoxia inhibits the growth and development of rats and induces cognitive dysfunction.High-level HIF-1α in chronic intermittent hypoxia indicates hypoxia-stress of the body, while compensatory increase of serum BDNF may be involved in neuronal cell damage regulation.

Objective To investigate the feasibility of NR2B small interference RNA(NR2B siRNA)carried by water-soluble lipopolymer(WSLP)for treatment of neuropathic pain in rats.Methods One hundred healthy male SD rats weighing 180-200 g were randomly divided into 5 groups(n=20 each):normal control group (group C),sham operation group(group S),neuropathic pain group(group NP),group WSLP-NR2B siRNA (group W)and group WSLP-negative NR2B siRNA(group WN).Neuropathic pain was induced by partial ligation of sciatic nenre.WSLP-NR2B siRNA complex was formed by binding WSLP and NR2B siRNA.Normal saline.WSLP-NR2B siRNA complex and WSLP-negative NR2B siRNA 20μl were injected intrathecally after operation in NP,W and WN groups respectively.Mechanical withdrawal threshold(MWT)and thermal withdrawal duration (TWD)were measured before(baseline)and at 3,7,14 and 21 days after operation.Ten animals in each group were sacrificed on the 3rd day after operation and the lumbar segment(L4-6)of the dorsal root ganglia was removed for determination of the expression of NR2B mRNA and protein using RT-PCR and Western blot analysis.Results Sciatic nerve ligation significantly decreased MWT and prolonged TWD and increased NR2B mRNA and protein expression in group NP as compared with group C.WSLP-NR2B siRNA complex significantly reduced sciatic nerve ligation-induced hyperalgesia and decreased NR2B mRNA and protein expression in group W as compared with group NP.Conclusion WSLP not only mediates NR2B siRNA successfully and inhibits the expression of NR2B,but also reduces neuropathic pain in rats.

Objective To detect radiation-induced changes of temporal lobe normal-appearing white matter on conventional MRI following radiation therapy (RT) for nasopharyngeal carcinoma (NPC).Methods The clinical and imaging features of 75 patients with nasopharyngeal carcinoma were retrospectively analyzed,all patients were confirmed by biopsy.All patients performed conventional MRI and Diffusion-tensor imaging (DTI) examinations,and there was no abnormal finding on conventional MRI.Eighteen patients without radiotherapy were selected as the control group and fifty-seven patients with radiotherapy were as the experimental group.We divided the experimental group into five subgroups based on completion time of RT:group 1 (less than 3 months,n =16),group 2 (3 to 6 months,n =12),group 3 (6 to 9 months; n =10),group 4 (9 to12 months,n =8),and group 5 ( more than 12 months,n =11 ).The mean diffusivity ( MD),apparent diffusion coefficient ( ADC ),fractional anisotropy ( FA),radial diffusivity ( λ⊥ ) and axial diffusivity ( k ‖ ) were calculated in bilateral temporal lobe.One-way analysis of variance (one-way ANOVA) test was used for comparison among groups.Results The mean λ⊥ values of the control group and experimental groups ( group1-5 ) after radiotherapy were ( 6.075 ± 0.341 ) × 10 -4 (6.700±0.379) × 10-4,(6.976 ±0.527) ×10-4,(6.621 ±0.388) ×10-4,(6.751 ±0.460) ×10-4,(6.222 ±0.256) × 10-4 mm2/s,respectively.The mean λ ‖ values of the control group and experimental groups were (12.524±0.713) ×10-4,(11.764 ±0.574) ×l0-4,(11.842±0.471) ×10-4,(11.569 ± 0.552) × 10-4,( 12.050 ±0.614) × 10-4,( 12.100 ±0.529) × 10-4 mm2/s,respectively.The mean FA values of the control group and experimental groups were 0.452 ± 0.030,0.379 ± 0.028,0.382 ± 0.028,0.389 ± 0.032,0.388 ± 0.022,0.423 ± 0.232,respectively.The three indicators were significantly different among groups ( F =10.485,4.625,16.539,respectively,P ＜ 0.05 ). Multiple comparisons showed that λ⊥ increased significantly in group 1-4 compared with that in the control group.In group 5,λ ⊥ was not significantly different from that in the control group,λ ‖ decreased in group 1-3 compared with that in the control group,but was not significantly different in the control group and group 4-5. In all experimental groups,FA decreased significantly. MD and ADC values in experimental groups were not significantly different from those in the control group. Conclusion Diffusion tensor imaging is a noninvasive and quantitative method to detect the structural changes in WM after RT and can provide scientific evidence for the early diagnosis and intervention treatment of radiation-induced changes.

Objective To detect specific cell killing effect of radiation combined with horseradish peroxidase (HRP)/indole-3-acetic (IAA) suicide gene therapy controlled by a novel radio-inducible and cancer-specific chimeric gene promoter in lung cancer. Methods We constructed a plasmid expressing HRP enzyme under the control of chimeric human telomerase reverse transcriptase (hTERT) promoter carrying 6 CArG elements, a plasmid expressing HRP enzyme under the control of hTERT promoter carrying single CArG element, and two control plasmids, which named pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc, respectively. After radiation, the proliferation inhibition and apoptosis induction effect of each type of plasmid in lung cancer cells (A549, SPC-A1) and normal lung cells (hEL) was detected by cell counting and Annexin V-FITC staining. The change of radiosensitivity of lung cancer cells with plasmid system was also detected by clonogenic assays. Results After a single dose radiation of 6 Gy,the average proliferation inhibition rates of pE6-hTERT-HRP, phTERT-HRP, pControl-HRP, and pControl-luc systems were 72. 92% ,40.60% , 51.00% and 25.19% (F= 67.31 , P< 0.01) in A549 cells ,64.63%,30.02%,48.23% and 23.16% (F=64.94, P< 0.01) in SPC-A1 cells, and 20.81%,18.05%, 44.20% and 18.32% (F=52. 19,P<0.01) in normal hEL cells, respectively. The average early apoptosis rates of these four plasmid systems were 36. 63%, 22. 30%, 24. 33% and 12. 53% (F =50. 99,P <0. 01) in A549 cells, 33.73%, 17. 37%, 22. 43% and 11.20% (F = 20. 76, P < 0. 01) in SPC-A1 cells, and 13.53 %, 12. 5%, 21.93% and 12. 16% (F = 15. 08, P < 0. 01) in normal hEL cells,respectively. The sensitizing enhancement ratios of the four plasmid systems were 3.45, 2. 29, 3.05 and 1.21 in A549 cells, while 2. 68, 2. 15, 3.05 and 1.21 in SPC-A1 cells, respectively. Conclusions The new suicide gene system controlled by chimeric promoter may provide a novel therapeutic modality for lung cancer.

Cordyceps sinensis is an expensive and tonic-valuable famous herb in China. Due to a narrow growing distribution, a low parasitic rate and a harsh living environmental condition, its wild recourse is not abundant. In re-cent years, with the improvement of living standards, people’s awareness of health care has increased strongly. That is because of people’s over-excavation need;and the natural resources are becoming increasing scarcity. In order to meet the demand for Cordyceps, domestic researchers conducted vigorous studies. Since 1987, four or five cul-tured Cordyceps species have been developed, namely fermented Cordyceps sinensis. Through subsequent unceasing research, they were put into a lot of production, bringing a certain benefits for income and society. Currently, there are four commercial brands on the market. They are Bailing, Jinshuibao, Xinganbao and Ningxinbao. This paper summarized the studies of its chemical compositions and clinical applications on fermented Cordyceps militaris in the recent five years. It will let scholars understand its value of the cultured Cordyceps militaris more comprehen-sively and supply more information for scholars in its further research.

Objective:To establish a 180-day mortality predictive score based on frailty syndrome in elderly sepsis patients [elderly sepsis score (ESS)].Methods:A prospective study for sepsis patients aged 60 years and above who were admitted to a medical intensive care unit of the General Hospital of Southern Theatre Command from January 1st, 2018 to December 31st, 2018 was conducted. Univariate analysis was performed on 19 independent variables including gender, age, body mass index (BMI), tumor, charlson comorbidity index (CCI), activity of daily living (ADL), instrumental activity of daily living (IADL), mini-mental state examination (MMSE), geriatric depression scale (GDS), clinical frail scale (CFS), sequential organ failure assessment (SOFA), Glasgow coma scale (GCS), acute physiology and chronic health evaluation (APACHEⅡ, APACHEⅣ), modified NUTRIC score (MNS), multiple drug resistance (MDR), mechanical ventilation (MV), continuous renal replacement therapy (CRRT) and palliative care. Continuous independent variables were converted into classified variables. Multivariate binary regression analysis of risk factors was conducted to screen independent risk factors which affecting 180-day mortality in elderly sepsis patients. Then a 180-day mortality predictive score was established, and the discrimination of the mortality of patients using CFS, SOFA, GCS, APACHEⅡ, APACHEⅣ, MNS scores were compared.Results:A total of 257 patients were enrolled, with a 180-day mortality of 60.7%. Univariate analysis showed that age, tumor, CCI, ADL, IADL, MMSE, CFS, SOFA, GCS, APACHEⅡ, APACHEⅣ, MNS, MDR, MV, CRRT, palliative care were risk factors of 180-day mortality in elderly sepsis patients [age: odds ratio ( OR) = 1.027, 95% confidence interval (95% CI) was 1.005-1.050, P = 0.018; tumor: OR =2.001, 95% CI was 1.022-3.920, P = 0.043; CCI: OR = 1.193, 95% CI was 1.064-1.339, P = 0.003; ADL: OR = 0.851, 95% CI was 0.772-0.940, P = 0.001; IADL: OR = 0.894, 95% CI was 0.826-0.967, P = 0.005; MMSE: OR = 0.962, 95% CI was 0.937-0.988, P = 0.004; CFS: OR = 1.303, 95% CI was 1.089-1.558, P = 0.004; SOFA: OR = 1.112, 95% CI was 1.038-1.191, P = 0.003; GCS: OR = 0.918, 95% CI was 0.863-0.977, P = 0.007; APACHEⅡ: OR = 1.098, 95% CI was 1.053-1.145, P < 0.001; APACHEⅣ: OR = 1.032, 95% CI was 1.020-1.044, P < 0.001; MNS: OR = 1.315, 95% CI was 1.159-1.493, P < 0.001; MDR: OR = 2.029, 95% CI was 1.197-3.437, P = 0.009; MV: OR = 6.408, 95% CI was 3.480-11.798, P < 0.001, CRRT: OR = 2.744, 95% CI was 1.529-4.923, P = 0.001, palliative care: OR = 5.760, 95% CI was 2.177-15.245, P < 0.001]. By binary regression analysis, CFS stratification ( OR = 1.934, 95% CI was 1.267-2.953, P = 0.002), MV ( OR = 4.531, 95% CI was 2.376-8.644, P < 0.001), CRRT ( OR = 2.471, 95% CI was 1.285-4.752, P = 0.007), palliative care ( OR = 6.169, 95% CI was 2.173-17.515, P = 0.001) were independent risk factors of 180-day mortality in elderly patients with sepsis. The model of "ESS = 0.660×CFS stratification+1.511×MV+0.905×CRRT+1.820×palliative care" was established. Receiver operating characteristic curve (ROC curve) analysis showed that the area under the ROC curve (AUC) for predicting 180-day mortality by ESS was 0.785 (95% CI was 0.730-0.834, P < 0.001). When the best cut-off value was 2.2 points, its sensitivity was 78.9%, specificity was 70.3%, the positive predictive value was 80.4%, and the negative predictive value was 68.3%. Simplified ESS was defined as "0.5×CFS stratification+1.5×MV+1×CRRT+2×palliative care". ROC curve analysis showed that AUC for predicting 180-day mortality by simplified ESS was 0.784 (95% CI was 0.729-0.833, P < 0.001). When the best cut-off value was 2.0 points, sensitivity was 76.9%, specificity was 70.3%, the positive predictive value was 80.0%, and the negative predictive value was 66.4%. Compared with CFS, SOFA, GCS, APACHEⅡ, APACHEⅣ and MNS, ESS had a significant difference in discriminating 180-day mortality in elderly patients with sepsis (AUC was 0.785 vs. 0.607, 0.607, 0.600, 0.664, 0.702, 0.657, 95% CI: 0.730-0.734 vs. 0.537-0.678, 0.537-0.677, 0.529-0.671, 0.598-0.730, 0.638-0.766, 0.590-0.725, all P < 0.05). Conclusions:CFS, MV, CRRT, and palliative care are independent risk factors of 180-day mortality in elderly patients with sepsis. We established ESS based on these risk factors. The ESS model has good discrimination and can be used as a reference and assessment tool for prediction and treatment guidance in elderly patients with sepsis.

Objective To investigate the P2X3 receptor expression in L6-S1 dorsal root ganglion (DRG)and bladder detrusor in a rat model of neurogenic bladder and urethra. Methods Eighty Sprague-Dawley rats wererecruited and randomly divided into a sacral injury group, a suprasacral injury group and a control group. Spinal tran-section was performed to establish the animal model of neurogenic bladder and urethra in rats of the sacral injurygroup and suprasacral injury group. Check the P2X3 receptor expression in DBG and bladder detrusor among thethree groups by Western blot test at 20 days after model establishment. Results P2X3 receptor expression in L6-S1DRG of sacral injury group was significantly less than that of the suprasacral injury group, which was in turn signifi-cantly higher than that of the control group. P2X3 receptor expression in bladder detrusor of sacral injury group wassignificantly lower than that of the suprasacral injury group, which was in turn significantly higher than that of thecontrol group. Conclusion There was close relationship between P2X3 receptor expression and dysfunction of blad-der and urethra.

Objective To explore the construction method and physicochemical properties of disulfide?bonded hyaluronic acid?functionalized sodium?meter probe for hepatocellular carcinoma, and its biological evaluation in vitro and feasibility of MRI. Methods Synthesis of hyaluronic acid?disulfide?bonded?poly ε?caprolactone (HA?SS?PCL) by disulfide?bonded alkynyl?terminated polycaprolacton (alkyne?SS?PCL) and azido?terminated hyaluronic acid (HA?N3) by clicking chemical reaction, then doxorubicin (DOX) and superparamagnetic iron oxide (SPIO) were encapsulated in HA?SS?PCL core by dialysis method.HA?SS?PCL@DOX/SPIO was prepared and its particle size,DOX and SPIO loading rate were measured. With PBS as control group, the safety of HA?SS?PCL on human hepatocellular carcinoma cells HepG2 and normal liver cells LO2 was evaluated by the methylthiazolyl tetrazolium (MTT) assay, and the cytotoxicity of HA?PCL@DOX/SPIO and HA?SS?PCL@DOX/SPIO on human hepatocellular carcinoma cells HepG2 was evaluated. Immunofluorescence and flow cytometry were used to observe the expression of CD44 receptor on the surface of HepG2 cells in HA?PCL@DOX/SPIO and HA?SS?PCL@DOX/SPIO groups. Through in vitro MRI, PBS was used as the control group to observe the changes of T2 signal intensity of HA?PCL@DOX/SPIO and HA?SS?PCL@DOX/SPIO groups when SPIO concentration was 10, 20, 40, 80 μg / ml. One way ANOVA test and t test were used. Results HA?SS?PCL@DOX / SPIO sodium?meter probes were successfully constructed. The particle size of HA?SS?PCL@DOX/SPIO was (126.9±6.3) nm,and they were spherical with uniform size. The loading rates of DOX and SPIO were 61.4% and 58.7%. MTT assay showed that the survival rate of HepG2 and LO2 cells was more than 80% even at 500 μg/ml of HA?SS?PCL, 66.6% in HA?PCL@DOX/SPIO group and 55.2% in HA?SS?PCL@DOX/SPIO group. Immunofluorescence and flow cytometry showed that HA?PCL@DOX/SPIO and HA?SS?PCL@DOX/SPIO groups all have strong fluorescence, and the latter has stronger fluorescence intensity the former fluorescence intensity was 139.70±8.52,less than the latter 245.06±13.21. In vitro MRI showed that the T2 signal intensity of HA?PCL@DOX/SPIO and HA?SS?PCL@DOX/SPIO was significantly lower than that of the control group (F values were 613.591 and 569.234,P=0.000), the latter decline rate was more significant. Conclusion The disulfide?bonded hyaluronic acid?functionalized sodium?meter probe for hepatocellular carcinoma has excellent physicochemical properties, good targeting and MRI functions on human hepatoma cell HepG2 at the cellular level in vitro.

Objective To preliminarily explore the anti?cancer efficiency of disulfide?bonded hyaluronic acid?functionalized targeted sodium?meter probe for hepatocellular carcinoma and the feasibility of MRI. Methods Twenty?one nude mice models of subcutaneous liver cancer transplantation were randomly divided into saline, hyaluronic acid?poly ε?caprolactone@ doxorubicin/superparamagnetic iron oxide (HA?PCL@DOX/SPIO) and HA?disulfide?bonded?PCL@DOX/SPIO (HA?SS?PCL@DOX/SPIO) groups, with 7 mice in each group. The experimental groups were injected with micelles at a dose of Fe 5 mg/kg through the tail vein, and the control group was injected with the same amount of saline via the tail vein. MRI was performed before and after injection (2 h, 4 h, 8 h). The T2 value of the region of interest (tumor) was measured and its decline rate was calculated. Twenty?one nude mice models of orthotopic liver cancer transplantation were randomly divided into saline group,HA?PCL@DOX/SPIO and HA?SS?PCL@DOX/SPIO groups, with 7 mice in each group. The experimental groups were injected with micelles at a dose of DOX 2 mg/kg through the tail vein by three consecutive times a day apart, and the control group was injected with the same amount of saline through the tail vein. Continuous observation for 15 days to calculate tumor inhibition rate. One way ANOVA test was used. Results The T2 value of HA?SS?PCL@DOX/SPIO group decreased significantly after 2, 4 and 8 hours (P<0.05) than initial time, which was distinct compared with HA?PCL@DOX/SPIO group. The growth rate of tumor in HA?SS?PCL@DOX/SPIO and HA?PCL@DOX/SPIO groups was significantly lower than that in the control group (F=21.513,P<0.05). The former had the most obvious inhibitory effect on orthotopic liver cancer (47.7% and 28.2%). Conclusion Disulfide?bonded hyaluronic acid?functionalized targeted sodium?meter probe for hepatocellular carcinoma(HA?SS?PCL@DOX/SPIO) has high anti?cancer efficiency and imaging function at the animal level in vivo, and can be used to monitor the early therapeutic effect of tumor at the molecular imaging level.

Objective:A multi-center and large sample volume study was conducted on the verification and improvement of the early established criteria for intelligent routine urinalysis validation (including the microscopic review rules and manual validation rules, referred to as intelligent criteria for short), in order to improve the clinical application of this intelligent criteria.Methods:A total of 31 456 urine specimens were collected from the inpatients and outpatients in six hospitals in China, from March to September 2019. Firstly, 3105 specimens were analyzed for preliminary verification and improvement of the intelligent criteria based on the results of the microscopic examination and manual validation. Secondly, 28 351 specimens were used to verify the clinical application of the improved intelligent criteria. All samples were manually validated as reference.Results:The approval inconsistency rate of the manual validation rules in the original intelligent criteria was 8.59% (202/2 352), and the interception inconsistency rate was 8.84% (208/2 352). The false negative rate and the microscopic review rate of the microscopic review rules were similar to the previous results. Based on an in-depth analysis of big data and the discussions by senior technicians from eight hospitals, one microscopic review rules and four manual validation rules were added, meanwhile two manual validation rule was deleted. The manual validation standards were unified. Finally, the intelligent criteria was improved. Based on the improved intelligent criteria, for microscopic review rules, the false positive rate, false negative rate (misdiagnosis rate), and microscopic review rate did not change significantly, which were 14.72% (457/3 105), 4.06% (126/3 105), and 24.73% (768/3 105), respectively. The approval inconsistency rate and the interception inconsistency rate of manual validation rules were both reduced to 0; the total manual validation rate of the intelligent criteria was 50.89% (1 580/3 105), and the auto-validation rate was 49.11% (1 525/3 105). The large sample volume verification results were consistent with the preliminary verification results of the improved intelligent criteria.Conclusion:This multi-center and large sample volume study had shown that the improved intelligent criteria had better clinical performance.