Acute Lung Injury ; therapy ; Humans ; Respiratory Distress Syndrome, Adult ; therapy

BACKGROUND: Severe acute respiratory syndrome (SARS) is caused by a genetically novel coronavirus that is caused by acute infectious disease. It is not yet clear for the immunology function of SARS patients in their convalescent stage.OBJECTIVE: To study the effects on T lymphocyte, and the titer profiling of 24 T cell receptor (TCR) V β subfamilies expressions in SARS convalescent patients.DESIGN: A self-control observation.SETTING: Central Laboratory, Guangdong Provincial Hospital of Traditional Chinese Medicine.PARTICIPANTS: Seventy-six cured SARS patients who received treatment in the Second Hospital Affiliated to Guangzhou University of Traditional Chinese Medicine between January and April 2003. All the patients corresponded to "clinical diagnostic criteria of atypical pneumonia", " diagnostic criteria of severe atypical pneumonia and discharge criteria" and "clinical diagnostic criteria and discharge criteria of severe acute respiratory syndrome". The involved patients, 30 male and 46 female, averaged (32±11 (years old. Another 10 subjects who simultaneously received health examination in the same hospital, 5 male and 5 female, aged (32±7(years, were involved in the study. Informed consents of detected items were obtained from all the subjects.METHODS：①Detecting the expression of 24 T cell receptor(TCR)V β subfamilies in SARS convalescent patients：Peripheral blood(2 mL) was collected from the healthy convalescent subjects,and EDTA-K2 was used as anticoagulant．In the flow cytometry delection tubes．10 μL various fluorescein-labeled mAb，such as anti-CD3,anti-CD4，anti-CD8,anti-CD25，anti-CD28，anti-HLA-DR，anti-CD3mAb conjugated with PC5，TCR Vβ1(PE+FITC)．Vβ2(PE+FITC)。Vβ3 (FITC)，Vβ4(PE+FITC)，Vβ5．1(PE+FITC),Vβ5．2(PE),Vβ5．3(PE)，Vβ7．1(PE+FITC)，Vβ7．2(FITC),Vβ8(FITC),Vβ9 (PE)，Vβ11(PE)，Vβ12(FITC)，Vβ13．1(PE),Vβ13．2(PE),Vβ13．6(PE+FITC)，Vβ14(FITC),Vβ16(FITC),Vβ17 (PE+FITC)，Vβ18(PE)，Vβ20(FITC)，Vβ21．3(FITC),Vβ22(PE+FITC)and Vβ23(PE)，was added in special flow tubes,and then 50 μL whole blood was added．The mixed solution was incubated away from light for 15 minutes．After erythrocytolysin being added，mixed solution was washed．Finally．cell deposit was dissolved in 300 μl phosphate buffer solution (PBS)．Coulter ESP flow cytometer was used for detection．For the analysis of TCR expression,an electronic gate was set on these cells and at least 5000 events per sample were collected．Three-color cytofluodmetric analysis was performed using a Coulter ESP flow cytometer．②Detecting the T cell subset,activated T and B cells,and the percentage of Ts and Tc cells：5000 cells were collected and used to calculate the expression of T cells (CD3,CD4 and CD8),the activated T and B cells(CD3+／CD25+,CD3+/HLA-DR+ and CD3-／HLA-DR+),as well as the percentage of Ts and Tc cells by Coulter ESP flow cytometer and its software．MAIN OUTCOME MEASURES：①The change of T cell subset(CD3，CD4，and CD8)from SARS convalescent patients．②The change of activated T and B cells(CD3+／CD25+，CD3+／HLA-DR+ and CD3-／HLA-DR+)．③The percentage of Ts and Tc cells(CD8+／CD28+，CD8+/CD28-)in convalescent patients．④Analysis of the 24 TCR V β subfamilies from SARS patients in convalescence．RESULTS：All data were explored to analyze the expression profiling of 24 TCR Vβ subfamilies,the data from 74 SARSpatients and 10 healthy controls were explored to other result analysis．①The detecting results of T celI subset：The percentage of CD4+T cell mean value was lower than the reference value[(33．33±6．64)％ vs．(43±9)％，P＜0．01]．The percentage of CD8+T cell mean value was higher than the refefence value[(34．07±6．40)％ vs．(30±9)％,P＜0．01]．② The expression of activated T and B cells：Percentage of HLA-DR+ T and B cell was Increased while the percentage of CD25+ T-cell was decreased compared with reference values．In 53 out of 74 patients,the percentage of CD25+ T cells was lower than the reference value,and 64 patients had a lower percentage in CD3+/CD25+ T cells．The percentages of CD3+/HLA-DR+ and CD3-／HLA-DR+ cells were higher than the normal reference value．T cells expressing higher CD3+／HLA-DR+ were found in 36 patients，and T cells expressing higher CD3-/HLA-DR+ were found in 30 patients．③The ratios of Ts and Tc cells：The percentage of Ts cells which expressed CD8+／CD28- was increased compared with reference value [(28．75±7．31)％ vs．(15．99±5．1)％，P＜0．01],while the percentage of Tc cells which expressed CD8+／CD28+ was decreased [(5．99±3．60)％ vs．(13．2±4．1)％，P＜0．01]．Thirty-nine patients were found to possess the lower Tc cells and forty-eight patients were found to possess the higher Ts cells．The ratios of both CD4+ and CD8+ T cells were in the normal reference value．④24 TCR Vβ subfamilies expressions in T cells：It was noteworthy that Vβ14 had a highest percentage in all 24 Subfamilies,and followed by Vβ 5．3,and Vβ 23 in the convalescent patients．The percentage of Vβ 14 was the highest in the normal controls,which was consistent with the results of SARS patients．But the other subfamilies expression patterns were different．There were significant differences between Vβ1,Vβ5．2,Vβ5．3,Vβ7．2，Vβ9，Vβ11,Vβ13．1,Vv13．2，Vβ17,Vβ18，Vβ22 and Vβ23．In the convalescent period，each TCR Vβ expression of SARS patients was higher than that of controls(P＜0．05-0．01)．CONCLUSION:In SARS convalescent patients,the increased CD8+CD28- T cell may elevate CD8+ T cell number;Meanwhile.the reduced CD3+ and CD4+ T cell number may be corresponding to the increased Ts cell number．For some inhibiting factor secreted by Ts cell was also increased．The usage pattern of 24 TCR Vβ subfamilies in SARS patients is different from that of control group．The increase of percentage of CD3+／HLA-DR+ and CD3-／HLA-DR+ T cell may be related to the late response of activated T and B cells.

Objective To explore the preparation of liposomal clodronate and investigate its inducing effects on the apoptosis of peritoneal macrophages in rats after severe acute pancreatitis (SAP).Methods Liposomal clodronate was prepared by means of thin film. SAP rat model was established by retrograde injection of 5% sodium taurocholate into the pancreatic duct. The peritoneal macrophages were obtained from SAP rats. After exposure to different doses of liposomal clodronate (50, 100,150 μl), the PM proliferation was determined by MTT colourimetry. The apoptosis of PM was measured by flow cytometry and agarose gel electrophoresis, respectively. Results The prepared liposomal clodronate had a suitable encapsulation efficiency of clodronate (5.8%) with an average size of 200 nm. The spherical shape of liposome was confirmed by transmission electron microscope. Exposed to liposomal clodronate of different doses resulted in a obvious growth depression (P＜0.01). The apoptotic rate of the PM was (10.32±0.34) %, (18.16±0.49)% and (29.87±0.35)% in three different dose groups and the difference was marked (P＜0.01). 1.2% of agarose gel electrophoresis of DNA extracted from apoptotic macrophages induced by liposomal clodronate showed clearer and characteristic ladder following the liposomal clodronate concentration. Conclusion Liposomal clodronate has a definite effect on peritoneal macrophages in SAP rats.

Objective To establish a HPLC method for determination of hypoxanthine and xanthine in Syngnathus.Methods A Lichrosper C_(18) Column was used.The mobile phase was methanol-0.1%HAc.Walvelength was 254nm.Results The linear range of hypoxanthine was within 5.91～94.56?g?mL~(-1),r=0.9999,sample recovery rate was 99.22%,RSD=(1.25)%.The linear range of Xanthine was within 2.04~32.64?g?mL~(-1),r=0.9998,sample recovery rate was 98.05%,RSD=1.21%.Conclusion This method has good repeatability and flexibility.It can be used for quality control in production of Syngnathus.

AIM: To establish a HPLC for determination of baicalin and emodin in Yiqing Capsule(Radix Scutellarial, Radix et Rhizoma Rhei). METHODS:A Kromasil C_(18) Column was used. The mobile phase was methanol -0.2% phosphoric acid for gradient elution. Baicalin and emodin were determined by dual wavelength, ?_1=315 nm, ?_2=266 nm. RESULTS:The linear range of baicalin was within 64.40-322.00 ?g?mL~(-1) (r=(0.999 8)). The average recovery was 99.96%,RSD was 1.57%(n=9). The linear range of emodin was within( 0.82)-13.20 ?g?mL~(-1)(r=0.999 9).The average revovery was 98.90%, RSD was 1.25%(n=9). CONCLUSION: This method has good repeatability and flexibility. It can be used for quality control in production of Yiqing Capsule. (Nan

Objective To develop a HPLC method for the determination of oridonin in the leaves and stems of Rabdosia nervosa.Methods All of reference substances and samples were separated with the mobile phase of methanol:water (45 ∶55) under isocratic elution for 20 min on a Kromasil C18 reversed-phase column(250 mm?4.0 mm,5 ?m),the flow rate was 1.0 mL?min-1,the detection wavelength was 235 nm,and the column temperature was 30 ℃.Results The content of oridonin was 0.047 mg/g in stems and 0.791 mg/g in leaves,the content in stems being about one seventeenth of that in leaves.The average recovery of oridonin was 98.33 %.Conclusion This method is simple,sensitive,accurate and suitable for quantitative determination of oridonin in Rabdosia nervosa.

AIM: To establish a HPLC for determination of puerarin and baicalin in Gegen Qinlian Micropill. METHODS: A Kromasil C 18 Column was used. The mobile phase was methanol-0.2% phosphoric acid. Puerarin and baicalin were determined by dual wavelength, ? 1=250nm. ? 2=315nm. RESULTS: The linear range of puerarin was within 32.0 ng—288.0 ng,r=0.9998 and the sample recovery was 100.17 % (RSD=1.00%(n=9)). The linear range of baicalin was within 52.6 ng～447.1 ng, r=0.9999 and the sample recovery was 100.06 % (RSD=1.06% (n=9)). CONCLUSION: This method has good repeatability and flexibility. It can be used for quality control of Gegen Qinlian Micropill.

In order to study the accumulation of Fritillaria thunbergii cultivar, peimine content in Xiaye, Kuanye, Duozi and Xiaosanzi bulbs of different sizes and parts was determined by HPLC-ELSE. The results indicated that the peimine content varied significantly with the cultivar type, the size and part of bulb. The distribution laws of peimine were as follow: Xiaosanzi > Duozi > Xiaye > Kuanye, small-size bulb > big-size bulb, core bud > scale. The peimine yield per plant in Duozi was the highest.
Cevanes ; analysis ; Chromatography, High Pressure Liquid ; Fritillaria ; chemistry ; growth & development

Objective:To investigate the clinical security and feasibility of neoadjuvant chemotherapy with imatinib following lap-aroscopy-guided intersphincteric resection for patients with gastrointestinal stromal tumor of the low rectum (GSTLR). Methods:Clini-cal data of nine patients with GSTLR who were admitted to the Shengjing Hospital between January 2007 and January 2011 were re-viewed. These patients were treated with neoadjuvant imatinib chemotherapy after laparoscopic intersphincteric resection. Results:Pri-or to neoadjuvant chemotherapy, the tumor diameter ranged between from 5 cm to 9 cm (median=7.0 cm). After imatinib chemothera-py, the tumor diameter decreased to 2-4.5 cm (median=3.5 cm, P<0.001). Laparoscopic surgery through intersphincteric resection was performed after imatinib treatment for 3-24 months (median=7 months). All patients received a protective stoma, which was closed 3 months after the surgery. The Wexner scale scores ranged from 1 and 4 (median=2) prior to neoadjuvant imatinib chemotherapy and changed to 1-5 (median=2) after the chemotherapy (P=0.397). After stomal closure operation, the scores significantly increased to 4-9 (median=7, P<0.001) but were not statistically significantly different from those before the therapy. One year after laparoscopic surgery, the Wexner scale scores ranged from 1 to 5 (median=2, P=0.842). Six patients were treated with imatinib for 24 and 30 months after lap-aroscopic surgery. Recurrence in pelvis occurred in only one patient, who ceased imatinib administration at the 30th month after the sur-gery. Conclusions: Laparoscopic surgery through intersphincteric resection was secure and feasible and thus could be used for treat-ment of GSTLR.

Objective To study the effect of infrasound on expression of calmodulin-dependent protein kinase II (CaMKII) and tau pro-tein in hippocampus of rats. Methods Fifty-six male Sprague-Dawley rats were randomized into control group (n=8), 1-day group (n=8), 7-day group (n=8) and 14-day group (n=32), and the 14-day group was subgrouped as 1-hour, 6-hour, 24-hour, 48-hour subgroups, naming after the time after infrasound exposure, 8 in each subgroup. All the test groups were put in an infrasound field with 8 Hz, 130 dB for 2 hours daily, while the control group was put in the infrasound instrument without infrasound exposure for 2 hours daily. The expression of pT286-CaMKII and tau protein in hippocampus was detected with immunohistochemisty, Western blotting and enzyme-linked immunoabsor-bent assay. Results The expression of pT286-CaMKII was the most in 14-day group (F>14.912, P<0.001), as well as the expression of tau pro-tein (F>36.229, P<0.001), and secondary in 7-day group (P<0.05). For 14-day group, the expression of tau protein was the most in 1-hour and 6-hour subgroups, and dropped down in 24-hour subgroup, although more than that in the control group (P<0.05). Conclusion Exposure of 8 Hz, 130 dB infrasound may induce phosphorylation of CaMKII and tau protein, and the expression of tau protein in hippocampal cells in rat, which may disturb their learning and memory function.