Objective To investigate the influence of hypoxia-reoxygenation on the myocardin gene expression of cultured rats' vascular smooth muscle cells(SMC).Methods The SMC were isolated from the media of the thoracic aorta vessel of Sprague-Dawley rats,and cultured with attachment-block manner.The morphology and cell counting of the cultured cells under hypoxia conditions were observed and comparedto that under normal culture condition.Total RNA extracted from the cultured cells,the expression of myocardin mRNA in SMC were measured at hypoxia status and at various time after reoxygenation through RT-PCR.Results The rats' vessel SMC was successfully cultured and showed a "peak-valley" shape.Self-made hypoxia equipment can produce a lower oxygen partial pressure without significant variation of pH value which met the experiment requirements in 2 4 hours .However,in the hypoxia conditions,the expression level of myocardin was lowest at the 12th hours,then increased.After 24 hours of hypoxia,the expression levels of myocardin began to increase at the 6th hour of reoxygenation and reached a normal level at the 12th hour of reoxygenation.Conclusions Hypoxia-reoxygenation has an effect on the expression of myodardin gene.

OBJECTIVE:To establish the method for the determination of tigecycline (TGC) in human plasma. METHODS:After precipitated by acetonitrile,the plasma sample was determined by LC-MS/MS. Using d9-TGC as internal standard,Kromasil C18 column was used with mobile phase consisted of water (containing 0.05% TFA)-acetonitrile (gradient elution) at flow rate of 0.6 ml/min,column temperature of 40 ℃. The ion transitions were performed under ESI positive MRM model at m/z 586.3→513.2 and m/z 595.3→514.3 for TGC and internal standard,respectively. RESULTS:The linear range of TGC was 25-2 000 ng/ml (r=0.999 8),and lowest quantification limit was 25 ng/ml;intra-day and inter-day RSD was 3.15%-7.23%,and relative error was-4.53%-10.48%. Plasma sample kept stable after 3 times of freezing and thawing cycle,at room temperature for 24 h,in automat-ic sample injector for 24 h and freezing for 42 d (RSD<15%). Plasma concentration of TGC was 0-438.0 ng/ml in one patient with pan-drug resistant bacteria infection(0-12 h after administration). CONCLUSIONS:The developed method is accurate,sensi-tive and specific,and can be used for plasma concentration determination of TGC and pharmacokinetic study.

Objective To measure the organ weights , blood physiological and biochemical parameters , and immune cells of BALB/cA-nu mice .Methods BALB/cA-nu mice at five and ten weeks of age were selected , and the organ weights , blood physiological and biochemical parameters were observed .The percentages of CD 3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were detected by FCM in BALB/cA-nu mice at six weeks of age in terms of its T, B lymphocyte cells and NK cell activity .Results At the same age, the weights of the body , liver, and kidney in male mice are significantly higher than females .The HGB、MCH、HCT in male mice are significantly higher than females (P <0.05).The weights of the lung, liver, and kidney, and the parameters of TP、GLOB、CHO、TBIL、UN are significantly lower at five weeks of age comparing with ten weeks (P <0.05).The BALB/cA-nu mice lacks T cells(0.18 ±0.06)%、CD4+T cells(0.26 ±0.08)%、CD8+T cells(0.13 ±0.04)%、The percentage of B cells is CD19+B cells (30.10 ±2.74)% and B220 +B cells(30.55 ±2.77)%.The percentage of NK cells is (1.35 ±0.29)%, and the percentage of granulocytes is (47.90 ±5.48)%.The BALB/cA-nu mice lack T cells and the group of cells .Conclusion The study suggests that the organ weights , blood physiological and biochemical parameters are affected by age and gender in BALB/cA-nu mice that lack T cells immune function .The studied parameters of BALB/cA-nu mice are similar with the same strain in other countries .

Objective To measure the organ weights , physiological and biochemical parameters and immune cells of NOD/SCID mice.Methods Mice at five and ten weeks of age were selected .The organ weights, blood physiological and biochemical parameters were observed .The percentages of CD 3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were checked by FCM in NOD/SCID mice at six week of age in terms of its T , B lymphocyte function and NK cell activity.Results Among the same sex group of NOD/SCID mice, the weights of kidney, liver, heart and lung, and the blood physiological indexes of RBC , HGB, HCT, MCV, MCH, RDW, and the blood biochemical indexes of TP , ALB, ALP, CHO, TBIL show significant difference between 5 and 10 weeks.At the same age, HCT, GLOB, A/G, CHO, TG, TBIL and the UN are significantly different between male and female NOD /SCID mice.NOD/SCID mice lack T cells (0.37 ±0.26)%、CD4+T cells (0.35 ±0.13)%、CD8 +T cells (0.47 ±0.10)%、CD19+B cells (0.13 ± 0.05)%、B220+B cells (1.20 ±0.44)%.The percentage of NK cells is (6.90 ±0.82)%, and the percentage of granulocytes is (47.88 ±15.54)%.Conclusions The study indicates that NOD/SCID mice show the deficiency of T , B and NK cells function .The organ weights , blood physiological and biochemical parameters are affected by age and gender . The studied parameters of NOD/SCID mice are similar with the same strain in other countries .

Objective To measure the organ weights , blood physiological and biochemical parameters , and immune function of Specific Pathogen Free ( SPF) Rag2 knockout ( KO) mice.Methods Rag2 knockout mice were selected at five and ten weeks , and the organ weights , blood physiological and biochemical parameters were observed .The percentages of CD3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were checked by FCM in Rag2 KO mice at six week of age in terms of its T , B lymphocyte function and NK cell activity .Results Among the same sex group of NOD/SCID mice, the weights of brain, lung, spleen, liver, heart, kidney, and the levels of TBIL, WBC in 10 weeks of age are higher than 5 weeks.At the same age, the weights of heart, kidney, liver, spleen, and the levels of AST, ALP, A/G, GLU, PLT, PCT, WBC, and LYM%in male mice are higher than females.The Rag2 KO mice lacks T cells(0.36 ±0.15)%、CD4+T cells(0.21 ±0.06)%、CD8+T cells(0.23 ±0.07)%、CD19+B cells(0.28 ±0.04)%、B220+B cells(2.03 ±0.42)%).The percentage of NK cells is(24.13 ±3.62)%, and the percentage of granulocytes is (57.20 ±3.85)%.Conclusion The study suggests that the organ weights , blood physiological and biochemical parameters are affected by age and gender in Rag2 KO mice, which main biological characteristics are similar with C 57BL/6J mice.The Rag2 KO mice show the deficiency of T , B cells function .

Objective To measure the organ weights , blood physiological and biochemical parameters of KK /Upj-Ay/J.Methods KK/Upj-Ay/J mice at five and ten weeks of age were selected , and the organ weights , blood physiological and biochemical parameters were observed .Results Parts of the organ weights , blood physiological and biochemical parameters of different ages and sexes were significant differences .The fasted blood glucose of KK/Upj-Ay/J mice reached 7.0mmol/L at 10 weeks of age .Conclusion The results show that the organ weights , blood physiological and biochemical parameters are affected by age and gender of KK /Upj-Ay/J mice.The fasted blood glucose reached the diabetes level at 10 weeks of age .

Objective To observe and compare the clinical efficacies between electroacupuncture and warm needling in treating low back pain.Method Seventy-eight eligible low back pain patients were randomized into group A of 28 cases, group B of 26 cases, and group C of 24 cases. Group A was intervened by electroacupuncture, group B was by warm needling, and group C was by medication. The short-form McGill Pain Questionnaire, Japanese Orthopaedic Association Scores (JOA), and Oswestry Disability Index were observed before and after treatment, and the therapeutic efficacies were compared.Result In group A, the McGill item scores [Sensory Pain Rating Index (S-PRI), Affective Pain Rating Index (A-PRI)] respectively after 1-week and 2-week treatment as well as in the 1-month and 3-month follow-up were significantly different from that before treatment (P<0.01,P<0.05). In group B and C, the McGill item scores after 2-week treatment and in the 1-month and 3-month follow-up were significantly different from that before treatment in the same group (P<0.01,P<0.05). The JOA and Oswestry scores were significantly changed respectively after 1-week and 2-week treatment and in the 1-month and 3-month follow-up in the three groups compared with that before treatment (P<0.05,P<0.01). After 1-week and 2-week treatment and in the 1-month and 3-month follow-up, the JOA and Oswestry scores in group A were significantly different from that in group C (P<0.05,P<0.01). In the 1-month and 3-month follow-up, the JOA scores in group B were significantly different from that in group C (P<0.05). The total effective rate was 85.7% in group A and 73.1% in group B, both significantly higher than 58.3% in group C (P<0.05). Conclusion Electroacupuncture and warm needling both can produce a significant efficacy in treating low back pain, but warm needling acts comparatively slowly and is less safe.

Objective To investigate the role of N-myc downstream regulated genes 2 (NDRG2) in attenuation of focal cerebral ischemic-reperfusin injury by sevoflurane preconditioning in rats.Methods Forty-eight healthy male SD rats weighing 280-320 g were randomly divided into 3 groups ( n =16 each):sham operation group (group S),ischemia-reperfusion injury group (group I/R) and sevoflurane preconditioning group (group Sev).Focal cerebral ischemia-reperfusion injury was induced by right middle cerebral artery occlusion (MCAO)for 120 min followed by 24 h reperfusion.In group Sev,2.0％ sevoflurane was inhaled 1 h once a day for 5 consecutive days at 24 h before MCAO.The neurologic function was evaluated at 24 h of reperfusion and than the rats were sacrificed,and the brain was removed for determination of infarct volume percentage,NDRG2 and activated Caspase-3 expression in ischemic penumbra by Western Blot and NDRG2 expression and location by immunohistochemistry.Results The infarct volume percentage,NDRG2 and activated Caspase-3 expression were higher,and neurologic function score was lower in groups I/R and Sev then in group S( P ＜ 0.05).The infarct volume percentage,NDRG2 and activated Caspase-3 expression were lower,and neurologic function score was higher in group Sev then in group I/R ( P ＜ 0.05).The intranuclear NDRG2 positive staining was decreased in group Sev than in group I/R.Conclusion Sevoflurane preconditioning can reduce focal cerebral ischemia-reperfusion injury by inhibiting the expression and activity of NDRG2 and apoptosis in rats.

Objective:To explore the clinical utility of metagenomic next-generation sequencing (mNGS) for patients with critically ill atypical rickettsial infections in the early diagnosis and therapy.Methods:From Jan 2020 to Aug 2022, clinical features, blood biochemical results, imaging data and mNGS results in patients with unexplained critical illnesses were collected and analyzed retrospectively. Fisher's exact test was used to compare the positive rate of mNGS and weil felix reaction.Results:All 15 patients with rickettsial disease had fever, 12 cases had headache, but only 3 had a typical rash or scab of diagnostic significance, 6 had septic shock and all had multi-organ dysfunction; blood mNGS tests were positive in 15 cases, of which 10 had Orientia tsutsugamushi detected in their blood and the remaining five had Rickettsia moschata detected in their blood. The positive rate of mNGS was significantly higher than that of the weil felix reaction (15/15 vs 0, P<0.001). All patients were given doxycycline and other treatments after diagnosis, of which 14 improved and were discharged, and one died 1 week after discharge due to critical condition and abandonment of treatment. Conclusion:mNGS can improve the detection rate of atypical rickettsiae in patients with negative routine test results, which can provide valuable reference basis for early diagnosis and early anti-infection treatment of patients with critical rickettsial disease.

Objective: To investigate the frequency of KRAS mutation in mucinous epithelial lesions of the endometrium, and analyze the correlation between KRAS mutation and the clinicopathologic features. Methods: The cohort included forty-three cases of mucinous epithelial lesions of the endometrium selected from July 2015 to October 2017 from Beijing Obstetrics and Gynecology Hospital, and 22 control cases. Genomic DNA was extracted from formalin-fixed paraffin-embedded tissue sections. Polymerase chain reaction amplification for KRAS exons 2 and 3 was performed, followed by sequencing using capillary electrophoresis. The Fisher exact test was used to compare the prevalence of KRAS mutation among the different groups. Results: The patients′age ranged from 33 to 77 years [mean (55.12±9.34) years, median 55 years]. None of the eight cases of endometrial hyperplasia with mucinous differentiation without atypia showed KRAS mutation. The frequency of KRAS mutations was 1/10 in endometrial atypical hyperplasia, 1/12 in endometrioid carcinoma, 4/11 in endometrial atypical hyperplasia with mucinous differentiation (EAHMD), 6/15 in endometrioid carcinoma with mucinous differentiation (ECMD) and 8/9 in mucinous carcinoma (MC), respectively. The differences were statistically significant between MC versus EC (P<0.01) and MC versus ECMD (P<0.05). Conclusion The high frequency of KRAS mutation in EAHMD, ECMD and MC indicates that KRAS mutational activation is implicated in the pathogenesis of endometrial mucinous carcinoma.