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Objective:To explore the characteristics of sleep disorders in patients with Parkinson's disease (PD) and its correlation with homocysteine.Methods:Totally 75 PD patients hospitalized in the department of neurology from January 2017 to June 2021 were selected and divided into sleep disorder group ( n=39) and non-sleep disorder group ( n=36)according to polysomnography, Parkinson's disease sleep scale(PDSS) and Epworth sleepiness scale(ESS). The basic clinical data, hematological examination results, scale evaluation data and polysomnography monitoring data of the above patients were collected during hospitalization to analyze the sleep characteristics of patients with Parkinson's disease and its correlation with homocysteine.SPSS 26.0 statistical analysis software was used for t test, Mann-Whitney U test, Pearson analysis, Spearman analysis and multivariate Logistic analysis. Results:The sleep efficiency (56.82±19.07)%, N2 phase ratio(48.67±17.70)%, N3 phase ratio(9.20%(19.00%)) and the leg movement micro-arousal index(0(1.20)) in the sleep disorder group were lower than those in the non-sleep disorder group (sleep efficiency (82.15±5.55)%, N2 phase ratio(57.02±2.80)%, N3 phase ratio(20.01%(3.93%)), the leg movement micro-arousal index(1.15(1.80)). The differences were statistically significant ( t/ Z=-6.087, -2.905, -3.773, -3.683, all P<0.05). The proportion of AHI (0.90(14.60)), N1 stage (19.50%(15.70%)), and periodic limb index (0(24.80)) in sleep disorder group were higher than those in non-sleep disorder group (AHI (0.60(0.30)), N1 stage (12.15%(3.15%)), and periodic limb index (0(0)). The difference was statistically significant ( Z=2.154, 5.250, 3.559, all P<0.05). The homocysteine (15.80(3.90) μmol/L), NMSS-insomnia correlation score (3.00(5.00)), MDS-UPDRS-Ⅰ(7.00 (10.00)), MDS-UPDRS-Ⅲ (23.00 (16.00)) in the sleep disorder group were higher than those in the non-sleep disorder group (homocysteine (14.10 (4.20)μmol/L), NMSS-insomnia correlation score (0(1.00)), MDS-UPDRS-Ⅰ(3.00 (2.00)), MDS-UPDRS-Ⅲ (17.00 (4.00)), and the differences were statistically significant( Z=2.557, 4.487, 2.952, 2.180, all P<0.05). The NMSS-olfactory correlation scores (2.00(4.00)) and PDSS (99.00 (40.00)) were lower than those in the non-sleep disorder group (NMSS-olfactory correlation scores (4.50 (7.00)) and PDSS (122.00 (28.00)), and the differences were statistically significant ( Z=2.450, 4.126, both P<0.05). Hcy was positively correlated with sleep disorder in PD patients ( r=0.297, P<0.05). Binariate logistic regression analysis showed that elevated homocysteine level might be a risk factor for sleep disorder in PD patients ( β=0.193, OR=1.213, 95% CI=1.029-1.430). Conclusion:Parkinson's disease patients with sleep disorder have the characteristics of sleep structure disorder, often accompanied by more serious motor disorders, and the olfactory function impairment is relatively mild. Elevated homocysteine levels may be a risk factor for sleep disorder in Parkinson's disease.
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As guided by the policy of building national regional medical centers, it is imperative to advance the construction of the national regional medical centers for mental health to achieve the sustainable and balanced development of the mental health cause of the country. The authors summarized the current status of national regional medical centers, and analyzed challenges faced in management and operation mode, government investment and compensation mechanism, expansion of high-quality psychiatric medical resources, as well as research and innovation of mental diseases of these centers. On such basis, the authors put forward corresponding countermeasures and suggestions for the next stage of development.
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Objective:To evaluate the early diagnostic value of tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and insulin-like growth factor binding protein-7 (IGFBP-7) in acute kidney injury induced by sepsis.Methods:A total of 85 sepsis patients admitted to the EICU and GICU in Shanghai East Hospital from September 2017 to June 2019 were divided into theAKI group ( n=37) and the non-AKI group ( n=48) according to KIDGO diagnostic criteria, and 20 healthy volunteers were served as the control group. The clinical data were recorded and samples of urine were collected at 0 h, 6 h, 12 h, 1 d, 3 d and 7 d post sepsis. The levels of TIMP-2 and IGFBP-7 in the urine were analyzed with ELISA at different time points. Based on the receiver operating characteristic curve (ROC) and the area under the curve (AUC), the early diagnostic value of urinary TIMP-2 and IGFBP-7 in sepsis-induced AKI patients was determined. Results:Compared with the control group, the levels of TIMP-2 and IGFBP-7 of the AKI group were significantly higher at the above time points ( P<0.05), while those of the non-AKI group showed no significant differences. The levels of TIMP-2 and IGFBP-7 of the AKI group were significantly higher than the those of the non-AKI group ( P<0.05). ROC analysis showed that when the AUC of urine TIMP-2 peaked at 1 d, the sensitivity and specificity reached 97.5% and 81.2%, separately with the cutoff value of 151.23 ng/mL. Furthermore, when the AUC of urine IGFBP-7 peaked at 12 h, the sensitivity and specificity reached 100% and 72.8%, separately with the cutoff value of 14.91 ng/mL. Interestingly, when the AUC of combined TIMP-2×IGFBP-7 peaked at 12 h, the sensitivity reached 98.0% and specificity reached 91.5% with the cutoff value of 2.09 [(ng/mL) 2/1 000]. There was no significant correlation between the levels of TIMP-2 and IGFBP-7 with SOFA and APACHEⅡ score at 1 d, 3 d and 7 d post sepsis in the AKI group ( P>0.05). Conclusions:Urine TIMP-2 and IGFBP-7 have early diagnostic value in sepsis-induced AKI. Besides, the combination of the two biomarkers have superior predictive value than each single of them.
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Objective:To explore the diagnosis and treatment strategies of primary ciliary dyskinesia (PCD).Methods:A 37-year-old male recipient who had repeated cough and sputum from childhood, with shortness of breath after activity and progressive development, the number of hospitalizations per year was ≥6 times, the MRC score was 2~3 diagnosed with primary ciliary dyskinesia through medical examination and multidisciplinary consultation, and received allogeneic double lung transplantation after medical treatment failure. Search related domestic and foreign literatures to explore and analyze the etiology, pathogenesis, clinical manifestations and imaging features, diagnosis and treatment of PCD.Results:PCD is an autosomal hereditary disease. Due to abnormal skeletal structure and/or functional development, clinical manifestations are mostly chronic sinusitis, bronchiectasis, otitis media and infertility. Chest imaging showed situs inversus totalis, right heart and diffuse cystic bronchiectasis infection with bronchial wall thickening, diagnosis depends on clinical manifestations, saccharin test, nasal nitric oxide test, high-speed video microscopy analysis, transmission electron microscopy, genetic testing and immunofluorescence.Conclusions:Lung transplantation is the only effective treatment for end-stage PCD. It not only saves patients' lives, but also significantly improves their quality of life.
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Objective To examine the expression profile of programmed death-ligand 1 (PD-L1) on lung endothelial or epithelial cells,and to determine the specific role of PD-L1 in mouse model of indirect acute lung injury (i-ALI).Methods Eighty male C57BL/6 mice were randomly divided into two parts (both n =40).The effects of different administration routes on the expression of PD-L1 were observed.The mice in each part were randomly divided into sham,i-ALI,i-ALI+small interfering RNA (siRNA) random sequence control,and i-ALI+PD-L1 siRNA which could specifically inhibit PD-L1 expression groups,with l0 mice in each group.i-ALI was reproduced in a mouse model of hemorrhagic shock in combination with a subsequent cecal ligation and puncture (CLP).In sham group,only bilateral femoral arteries were ligated without catheterization or bleeding,and only cecum was separated but perforation was not ligated.Intravenous or intratracheal delivery of PD-L1 siRNA was performed 2 hours following the resuscitation to suppress the expression of PD-L1 on lung endothelial or epithelial cells.The mice in i-ALI+siRNA random sequence control group were given siRNA random sequence without inhibition effect on PD-L1 expression,and those in sham group and i-ALI group were given 100 μL phosphate buffered saline (PBS).The mice were sacrificed at 24 hours after CLP,and samples of blood,lung tissue and bronchoalveolar lavage fluid (BALF) were harvested.Expressions of PD-L1 were determined with flow cytometry.Cytokines and chemokines in plasma,lung tissue and BALF were determined by enzyme linked immunosorbent assay (ELISA).The protein concentration in plasma and BALF and the activity of myeloperoxidase (MPO) in lung tissue were quantitatively measured.The pathological changes in lung tissue were observed under light microscope.Results ① Compared with sham group,PD-L1 expression on lung endothelial or epithelial cells were significantly elevated in i-ALI group [endothelial cells:(27.88 ± 1.53)％ vs.(19.64 ± 1.03)％,epithelial cells:(58.70 ± 8.21)％ vs.(29.23 ± 3.94)％,both P ＜ 0.05].② Mice received intravenous delivery of liposomal-encapsulated siRNA had significantly lower expression of PD-L1 on lung endothelial cells as compared with that of i-ALI group [(21.37 ± 0.76)％ vs.(27.88 ± 1.53)％,P ＜ 0.05].Intratracheal delivery of naked PD-L1 siRNA mainly inhibited the PD-L1 expression on epithelial cell as compared with that of i-ALI group [(31.23±4.71) ％ vs.(58.70±8.21) ％,P ＜ 0.05].The expression of PD-L1 in pulmonary microvascular endothelial cells or pulmonary epithelial cells of i-ALI mice was not affected by siRNA random sequence.③ PD-L1 silencing on pulmonary endothelial cells induced by intravenous delivery of PD-L1 siRNA led to a lower protein ratio of BALF/plasma [(4.48 ± 0.35) × 10-3 vs.(6.11 ± 0.56) × 10-3,P ＜ 0.05] and a decreased MPO activity in lung tissue (U · μg-1 · min-1:2.48 ± 0.47 vs.4.56 ± 0.52,P ＜ 0.05) as compared with that of i-ALI group.Moreover,inflammatory mediator levels such as interleukin-6 (IL-6),monocyte chemoattractant protein-1 (MCP-1),macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α) in lung tissue or plasma were significantly reduced following PD-L1 suppression on endothelial cells as compared with those of i-ALI group [IL-6 (ng/g):177.4±23.2 vs.287.9±57.3,MCP-1 (ng/g):839.6±91.7 vs.1 395.7±211.9,MIP-2 (ng/g):923.7± 107.3 vs.1 700.9±240.2 in lung tissue;IL-6 (ng/L):950.2±192.7 vs.1 828.2±243.6,TNF-α (ng/L):258.7±29.1 vs.443.0 ± 58.1,MCP-1 (ng/L):2 583.8±302.3 vs.4 328.1 ±416.4,MIP-2 (ng/L):1 512.9± 165.6 vs.2 005.9 ± 85.7 in plasma,all P ＜ 0.05],however,there was no significant change in the levels of inflammatory factors in BALF.It was shown in lung tissue histology that PD-L1 silencing on pulmonary endothelial cells induced by intravenous delivery of PD-L1 siRNA led to lessened pulmonary edema and reduced immune cells emigration.Intratracheal delivery of PD-L1 siRNA for PD-L1 suppression on epithelial cells had minimal effects on protein ratio of BALF/plasma,MPO activity,inflammatory mediator expressions in lung tissue,plasma,and BALF as well as lung tissue histology.Conclusion PD-L1 silencing on endothelial cells but not epithelial cells protected mice against hemorrhagic shock-sepsis induced i-ALI.
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Objective To investigate the influence of Joint Commission International (JCI) accreditation on nursing practice.Methods Totally 30 medical institutions which had passed the JCI accreditation were investigated to compare the scores of nursing practice before and after JCI accreditation.Results Before JCI accreditation,the average standardized score of governance,leadership and management dimension was 81.20,but the scores of other dimensions were below 80.After the JCI accreditation,the quality improvement and patient safety(QPS) dimension scored 81.10,and scores of other dimensions were all above 90.The differences of all dimensions were statistically significant (P＜0.001) before and after the JCI accreditation,the scores were all much greater than those before the accreditation,and improvement of AOP(77.37％)and IPSG(69.86％)was most obvious with statistical significance (P＜0.001).Conclusion JCI accreditation plays a certain role in promoting nursing practice,making nursing process more systematic and comprehensive and the safety of nursing guaranteed.
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Objective To investigate the effect of CT guided percutaneous targeted coblation nucleoplasty in treatment of cervical intervetebral disc herniation.Methods Fifty patients who were diagnosed with cervical intervetebral disc herniation by CT or MRI were received the treatment of CT guided percutaneous targeted coblation nucleoplasty and the follow-up check after operation.The effects according to Macnab therapeutic criteria and VAS method were evaluated and recorded before operation and after operation at the third day,2 weeks,3 months.Results All patients accepted follow-up three months after operation.The effects were assessed by Macnab therapeutic criteria:1 9 cases were excellent (37.5%),29 cases good (58.3%),2 cases bad (4.2%),and the effective rate was 92.5%.The VAS scores in three months after operation were significantly lower than that before operation (P <0.05 ). Conclusion Computed tomography guided percutaneous targeted coblation nucleoplasty is a safe,effective and minimally invasive method for treating cervical intervetebral disc herniation.
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Objective: To investigate the effects of fosinppril on myocardial cell apoptosis and apoptosis-associated gene expression in chronic heart failure (CHF) rats. Methods: CHF model was established by partially banding of abdominal aorta superior to renal artery. The experimental rats were randomly divided into 3 groups: Sham operation group and CHF group, the rats in both groups received stomach normal saline; Fosinopril group, the rats received stomach fosinopril 10 mg/kg?d.n=10 in each group and all animals were treated for 8 weeks. LVEDP, ±dp/dtmax and LVMI were examined, left ventricular myocardial cell apoptotic index (AI) was measured by TUNEL method, Bcl-2 and Bax protein levels were detected by immunohistochemistry and caspase-3 protein expression was assessed by Western blotting. Results: Compared with Sham operation group, CHF group had increased LVEDP, LVMI, AI, elevated protein expressions of Bax and Caspase-3,P Conclusion: Fosinopril may inhibit myocardial cell apoptosis which occurred during CHF, by up-regulating Bcl-2 expression and down-regulating expressions of Bax and Caspase-3 in CHF rats, therefore improve the cardiac function.
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Objective To explore the influence of network intervention on lifestyle and behaviors habit of the middle -aged patients with hypertension.Methods All 300 middle -aged patients with hypertension were randomly assigned into the study group and the control group.Each group had 150 cases.All patients in the both two groups were given hypotensive drugs according to treatment specification of hypertension.The patients in the study group received a network intervention for 2 years,while the patients in the control group received an outpatient service follow-up.The difference of lifestyle and behaviors habit of the patients in the two groups were observed and compared. Results There were no significant differences with all items of the lifestyle and behaviors habit of the patients between the two groups before intervention(P >0.05 ).In the 12th end of month after intervention,there were no significant differences between the two groups with the items of smoking control,working and resting regularly,hobby and psychological balance(P >0.05),but other items in the study group were significantly better than those in the control group(Z =-3.062,-2.509,-2.239,-2.056,-2.126,-2.045,-2.023,all P <0.05).In the 24th end of month after intervention,all items in the study group were significantly better than those in the control group (Z =-4.078,-3.792,-2.080,-2.098,-1.985,-2.478,-2.173,-2.478,-3.290,-2.514,-2.730,all P <0.05 ).Conclusion The network intervention can significantly improve lifestyle and behaviors habit of the middle -aged patients with hypertension.Its effect is much better than that of outpatient service follow -up.
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Sepsis induced by burns, trauma, and surgical stress, remains a major cause of death of patients in ICUs. A growing number of evidence shows that sepsis may result in dysfunction of innate and adaptive immune system, including the abnormal disorder of immune response of T lymphocytes, regulatory T lymphocytes, and dendritic cells, resulting in a state of immune depression. It is of great significance that dynamic monitoring of immune function and the related indicators might help to assess the risk of secondary infection, the prognosis of septic patients, and to guide the treatment of severe sepsis.
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Humans , Burns , Allergy and Immunology , Dendritic Cells , Allergy and Immunology , Intensive Care Units , Monitoring, Immunologic , Prognosis , Sepsis , Allergy and Immunology , T-Lymphocytes, Regulatory , Allergy and ImmunologyABSTRACT
Objective To explore effectiveness of combination of traditional Chinese medicine and western medicine therapy for patients with Barrett esophagus (BE).Methods Patients with BE were divided into the control group and observation group according to random number table.The observation group was received treatment of com-bination of argon plasma coagulation (APC)and traditional Chinese medicine,while the control group was received treatment of APC alone.Clinical effectiveness was observed at 3,6,12,18months after treatments,recurrence rate was observed 6,12,18months after treatments.Results Clinical effectiveness were significantly higher in the observation group than in the control group 3,6,12,18months after treatments (χ2 =4.71,3.55,7.28,5.10,all P <0.05). Recurrence rate were significantly lower in the observation group than in the control group 6,12,18 months after treat-ments (χ2 =5.84,6.51,all P <0.05).Conclusion Combination therapy of traditional Chinese medicine and west-ern medicine shows confirmed effects to BE.Low recurrence rate in addition makes it become a promising clinical therapy in treatment of BE.
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Objective: To explore the effects of ramipril, trimetazidine and the combination of ramipril and trimetazidine on renal cell apoptosis index (AI) and cytochrome C (Cyt-C) expression in experimental rats with chronic heart failure (CHF). Methods: CHF model was established by partially banding of abdominal aorta superior to renal artery in experimental rats. A total of 50 male Wistar rats were randomly divided into 5 groups: Sham operation group, Model group, Ramipril group, Trimetazidine group and Combination (ramipril and trimetazidine) group.n=10 in each group. Renal tubular cell AI was examined by TUNEL method, mRNA and protein expressions of Cyt-C were detected by RT-PCR and Western Blot analysis in each group respectively. Results: Compared with Sham operation group, Model group had increased AI of renal tubular cells, increased mRNA and protein expressions of Cyt-C,P Conclusion: Ramipril and trimetazidine can inhibit renal cell apoptosis and effectively improve the renal function in CHF rats. Combined medication is better than either of them alone.
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Objective To observe the influence of post-operative swallowing painwith Disposcope endoscope or Macintosh laryngoscope in patients with difficult visible glottis. Methods Thirty-six patients scheduled for elective surgery under general anesthesia whose glottis was not visible underMacintosh laryngoscope (according to Cormach-Lehane Grading of laryngoscopic view) were randomized into 2 groups (n = 18 each):Disposcope endoscope group (group D) and Macintosh laryngoscope group (group M). The rate of glottis exposure and successful intubation, swallowing pain at 30 min, 1, 3 d after extubation were observed. Results Compared with group M , the successful rate of glottis exposure and first intubation was significantly higher than thatin group D (P < 0.05). Swallowing pain VAS was higher in group M than in group D at 1d afterextubation (P < 0.05). Conclusions Compared with Macintosh laryngoscope , Disposcope endoscope in patients with difficult visible glottis could induce higher successful rate of glottis exposure and first intubation , shorter time of intubation and lower VAS of swallowing pain at 1d after extubation.
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Objective To evaluate the efficacy of Disposcope endoscope for tracheal intubation in patients with difficult glottis exposure.Methods Sixty adult patients of both sexes,aged 22-68 yr,scheduled for elective oro-tracheal intubation under general anesthesia,whose glottis was not visible under Macintosh laryngoscope (grade Ⅲ or according to Cormach-Lehane Grading of laryngoscopic view),were randomized into 2 groups (n =30 each) using a random number table:group Macintosh laryngoscope-assisted Disposcope endoscope (group DM) and group Disposcope endoscope (group D).The glottis exposure condition and time,intubation condition and time,assisted lifting mandible,laryngeal mucosa bleeding and postoperative sore throat and hoarseness were recorded.Results The success rate of intubation at second attempt achieved 100％ in the two groups.Compared with group DM,the incidence of assisted lifting mandible was decreased,and no significant changes were found in the success rate of glottis exposure,glottis exposure time,intubation time,success rate of intubation at first attempt,incidence of laryngeal mucosa bleeding,and postoperative sore throat and hoarseness in group D.Conclusion Disposcope endoscope can improve the condition for tracheal intubation significantly after lifting mandible in patients with difficult glottis exposure.
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With the rapid development of new media,medical students' ideological and moral qualities,physical and mental health have been unprecedentedly affected.Traditional ideological and political education modes for medical students encounter great challenges.In order to strengthen the ideological and political education for medical students,we need to focus on three aspects,namely enhancing information qualities of the subjects of ideological and political education,enhancing the supervision of public opinion and guidance on new media and building new interactive environment.
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Objective To conduct extensive quality control tests on Madin-Darby Canine Kidney ( MDCK) cells used for the production of influenza vaccine .Methods Tests for characteristics , extraneous agents, endogenous agents and tumorigenicity were performed on MDCK cells according to Chinese Pharma -copeia Book III .Cell lysate and DNA of MDCK cells were tested for oncogenicity in the light of new interna -tional requirements .Results The MDCK cells extracted from canis were adherent cells with an epithelial morphology, whose average number of chromosome was 80±1.No bacteria, fungi and mycoplasma contami-nation were detected . The detection for extraneous and endogenous virus showed that there was no nonspecific virus causing cytopathic effect , hemadsorption , hemagglutination or animal death .Tests for re-verse transcriptase , bovine viruses and canine viruses were all negative .Each nude mouse was injected with 107 viable cells to observe their tumorigenicity .Twelve weeks after cell injection , no node was found at the injection site and in large organs by gross anatomy .There was no significant difference between test group and negative control group .The test for tumorigenicity of viable cells was negative .Cell lysate and cellular DNA collected from equivalent amount of cells were respectively injected into nude mice , and no node forma-tion was found.There was no significant difference between the test cells and negative controls in pathology indicating that the tested MDCK cells were non-oncogenic .Conclusion It showed the possibility of using MDCK cells for the production of influenza vaccine .
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Objective To evaluate the quality of four domestic and three imported fourth-generation HIV diagnostic reagents.Methods The specificity and sensitivity of these assays were analyzed when testing HIV negative samples and HIV-1 RNA positive samples.The relative seroconversion sensitivity index was analyzed when testing BBI seroconversion panels.Results The sensitivity of seven 4th-generation assays were 100％ (95％ CI:99.86％-100％ ),and one sample at the window period of HIV-1 infection were detected as positive.Of the seven assays,one imported assay exhibited the relative largeδ + value (1.0892),and the small δ+ value were found on the remaining six assays (0.0836-0.3003 ).For the samples negative for HIV antibody,varying degrees of false positives were observed on the seven assays ( specificity:97.80％ -99.60％,δ- value:-1.3803 to -0.4778).When testing the BBI seroconversion panels,the relative seroconversion sensitivity index of domestic assays were -0.500-0,however,which of imported assays were -0.600 and -0.700.Conclusion The seven reagents exhibited high sensitivity and specificity.The 4th generation HIV assays can be used as blood screening reagents to find the samples at window period of HIV-1 infection,thus indicating the certain meaning in reducing the transmission risk of HIV-1 for fourth-generation HIV diagnostic reagents.However,the better efficiency to detect HIV-1 early infection was observed on the imported assays than on the domestic assays.
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@#ObjectiveTo discuss the perioperative nursing of sigmoid colon cystoplasty in neurogenic bladder patients after spinal cord diseases. Methods31 patients with neurogenic bladder due to the spinal cord diseases and received sigmoid colon cystoplasty were reviewed and followed up. ResultsThere were 2 cases complicated with bowel obstruction, 1 case with acute alleosis and 1 case with mycotic infection in bladder after the operation, who were all cured. After discharge, 2 cases lost the follow-up. During the follow-up period of 6 months to 4 years, 29 patients recovered well and kept in self-intermittent catheterization. ConclusionPreoperative psychological nursing and intestinal and bladder preparation, postoperative gastrointestinal and urinary drainage catheters nursing, intermittent catheterization training, regular follow-up and health guiding were the key nursing contents and the important step for ensuring an excellent outcome for the operation.
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Objective To investigate the effects of low-dose of hydrocortisone on circulating thymus-dependent lymphocyte (T lymphocyte) apoptosis in patients with septic shock. Method fifty-seven patients with septic shock admitted into ICU from January 2006 to January 2009 were prospectively randomized (random number) to treatment group and control group. Another 20 healthy volunteers and 18 patients with sepsis alone were included as external control groups.The patients of treatment group and control group were treated with low-dose of hydrocortisone and placebo,respectively. Samples of peripheral blood were taken from healthy volunteers and patients 0 hr,24 hrs,48 hrs,72 hrs and 168 hrs after onset of the disease to determine the circulating T lymphocyte apoptosis by using the assays of Annexin V and flow cytometry. Least significant difference t -test was used for multiple comparisons. Results The percentage of Annexin V-positive CD4+ T lymphocytes in the primary stage was (11.01 +4.52)% in septic shock patients, (4.41 + 1.45)% in healthy volunteers, and (7.87 + 3. 82)% in patients with sepsis alone. And in the initial setting, the percentage of Annexin V-positive CD4+ T lymphocytes in the septic shock patients was higher than that in healthy volunteers ( P < 0.05) and in patients with sepsis alone ( P < 0.05). The percentage of Annexin V-positive CD8 + T lymphocytes at the beginning was (11.33+19.62)% in septic shock patients, (9.62+8.32)% in healthy volunteers, and (13.09+ 15.84)% in patients with sepsis alone (P > 0.05 between three groups). The percentages of Annexin V-positive CD4+ T lymphocytes in control group after 24 his, 48 hrs and 72 hrs were(13.51+6.85)%, (19.39 + 6.63)% and (15.33+ 6.21)%, respectively. And the percentages of Annexin V-positive CD4+ T lymphocytes in treatment patients after 24 hrs, 48 hrs and 72 hrs were (17.4 + 7.21)%, (22.61 + 5.64)%, and (25.73 + 6.91)%, respectively. The percentage of Annexin V-positive CD4+ T lymphocytes in septic shock patients was higher than that in control groups ( P < 0.05). The percentages of Annexin V-positive CD8+ T lymphocytes in control group after 24 hrs, 48 hrs and 72 hrs were (11.49+ 11.73)%, (12.74+ 10.39)% and (13.28+ 16.6)%, respectively, and in the treatment group, those were (9.49 + 8.9)%, (15.32+18.17)% and (13.68+16.84)%, respectively (P >0.05 between two groups). In the meantime, the percentages of Annexin V-positive CDS'1' T lymphocytes in control group and in treatment group were (12.72+ 19.69)% and (13.88 + 13.28)%, respectively (P >0.05). Conclusions Low-dose of hydrocortisone could induce CD4+ T lymphocyte apoptosis and has no effects on CD8+ T lymphocyte apoptosis when it is used to treat septic shock.
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Objective To pan and characterize anti-HIV-1 Fab by the phage antibody library technology. Methods Total RNA were extracted from lymphocytes which were isolated from peripheral blood collected from asymptomatic HIV-1 infected donors with high titer antibody against HIV-1. The genes of heavy chains Fd fragment and light chains of antibody were amplified by RT-PCR. The phagmids pComb3X cloned Fd and light chain genes were transformed into E. coli XL1-Blue by electroporation to construct phage Fab library. By three runs of "absorption-elution-neutralization-enrichment", the clones were induced by IPTG and characterized by ELISA. The positive clones were sequenced and analyzed the sequences. Subsequently, Fab antibodies of these positive clones were induced to expressed and purified, then the recombinant virus neutralization assay was performed. Results A phage Fab library was constructed with 8×106 members, and 11 positive clones were obtained by detecting IPTG-induced-expressing Fabs with ELISA. By analysis of the sequences, 10 light chain genes and 8 Fd genes were ensured to be obtained. Compared with the genes of anti-HIV-1 antibodies in HIV sequence database, the gene sequences we obtained were highly homologous to some patent genes of anti-HIV-1 gp120 antibodies in HIV sequence database( light chains with 60%-90% identity, Fd with 71%-85% identity); The CDRs of these positive clones were determined by comparing the positive clone genes with antibodies' genes in V base database, furthermore, CDRH3 of these positive clones has the length of 12-22 aa. Strand shift had little effect to improving affinity of our Fab clones. Fab antibodies were induced to express at the concentration of ＞ 10 mg/L. Three Fab antibodies neutralize HIV-1 virus to some extent. Conclusion The studies will provide the basis on further study on the anti-HIV-1 Fabs obtained successfully.