Recognition and progress of idiopathic intracranial hypertension / 中国医师杂志

Idiopathic intracranial hypertension (IIH) is a syndrome of unexplained increased intracranial pressure with normal cerebrospinal fluid and without organic brain lesions. The etiology and pathogenesis of IIH remain unclear, and IIH patients may develop irreversible visual impairment. At present, there are no guidelines and expert consensus on diagnosis and treatment of IIH in China. This review aims to introduce the pathogenesis, diagnosis and treatment of IIH, in order to help clinicians improve their understanding of the disease and to identify, diagnose and treat IIH as early as possible, and improve the prognosis of patients.

Impact of surgical treatment for locally recurrent rectal cancer on the quality of life of patients / 中华胃肠外科杂志

Objective: To investigate the impact of surgical treatment on quality of life in patients with locally recurrent rectal cancer (LRRC). Methods: A descriptive case series study was performed. The complete clinical data of 62 patients who met the diagnostic criteria of LRRC and treated by surgical procedures in Huashan Hospital of Fudan University from January 2012 to November 2019 were analyzed retrospectively. All the patients were followed up at least 12 months. Assessments of urinary function, sexual function, mobility function of lower limb and quality of life were documented. Patients with distant metastasis and surgical history of the urinary system were excluded. According to the criteria of Memorial Sloan Kettering Cancer Center (MSKCC), recurrence were divided into central (n=27), anterior (n=20), posterior (n=7), and lateral (n=8) subtypes. Baseline characteristics, surgical procedures and short-term complications were analyzed. International prostate symptom score (IPSS) and grade of voiding dysfunction were used to evaluate the urinary function. Higher score of IPSS and higher grade of voiding dysfunction indicated worse voiding function. Sexual function for both genders was assessed preoperatively and postoperatively. International index of erectile function-5 (IIEF-5) was used for assessment of male patients and higher score indicated better function. Female sexual function index (FSFI) was used in females and higher score indicated better function. Short-form health survey with 36 items (SF-36), yielding an 8-scale profile of functional health (physical functioning, role-physical, bodily pain, general health, vitality, social functioning, emotional health and mental health) was used to evaluate the quality of life. The higher score indicated the better quality of life. Results: All the operations of 62 patients completed successfully and R0 resection rate was 88.7% (55/62). Postoperative surgical complications occurred in 16 cases (25.8%), including 3 patients of Clavien-Dindo classification III. At postoperative 3-month, 42 patients without ileum cystectomy or ureterostomy suffered from different grade of voiding dysfunction. IPSS increased significantly after the surgery (before surgery: 12.36±4.75, after surgery: 18.40±4.77, t=-9.128, P<0.001). There was no significant difference among the subtypes (P>0.05). At postoperative 12-month, IIEF-5 decreased from 14 (0~25) to 9 (0~19) in males (Z=-5.174, P<0.001) and FSFI deceased from 8.4 (2.0-27.0) to 2.0 (2.0-18.4) in females (Z=-3.522, P<0.001). Scores of physical functioning and role-physical decreased significantly [physical functioning: before surgery 70 (35-85), after surgery 65 (30-80), Z=-3.685, P<0.001; role-physical: before surgery 50 (0-50), after surgery 25(0-75), Z=-4.065, P<0.001], while those of social functioning role-emotional and mental health increased significantly after the surgery [social functioning: before surgery 44 (22-78), after surgery 56 (0-89), Z=-3.509, P<0.001; role-emotional: before surgery 17 (0-100), after surgery 33 (0-100), Z=-2.439, P=0.015; mental health: before surgery 40 (36-76), after surgery 52 (24-80), Z=-3.395, P<0.001]. All surgical procedures decreased the voiding function of LRRC patients and the sexual function of male patients (all P<0.01). However, only total pelvic exenteration and posterior pelvic exenteration decreased FSFI in female patients [before surgery: 8.4 (2.0-27.0) after surgery: 2.0 (2.0-18.4), Z=-2.810, P=0.005]. Conclusions: Multi-visceral resection in LRRC patients may damage voiding and sexual function. However, successful and effective surgical treatment can improve the psychosocial health of LRRC patients.

Postoperative deep vein thrombosis in patients after selective ankle or mid-hindfoot surgery / 中华创伤骨科杂志

Objective:To investigate the prevalence of deep vein thrombosis (DVT) in patients after selective ankle or mid-hindfoot surgery.Methods:A retrospective analysis was conducted of the 109 patients with ankle or mid-hindfoot disease who had been treated from January 2018 to December 2019 at Department of Orthopaedics, Wuhan Fourth Hospital. They were 65 males and 44 females, aged from 32 to 74 years (average, 49.0 years). Ultrasonography was performed at preoperative 1 day, postoperative 2 and 6 weeks to determine the occurrence, location and clinical symptoms of DVT. The patients were divided into an early DVT group, a late DVT group and a DVT-free group according to the occurrence and onset time of DVT. The 3 groups were compared in terms of gender, age, body mass index and tourniquet duration.Results:The incidence of postoperative lower limb DVT was 22.9% (25/109). All the thromboses were observed beyond the distal plane of the popliteal vein. 72.0% of the DVT patients were clinically asymptomatic. There was no significant difference in gender, age or body mass index between early DVT group ( n=17), late DVT group ( n=8) and DVT-free group ( n=84) ( P>0.05). The incidence was 68.0% (17/25) for early DVT and 32.0% (8/25) for late DVT. The intraoperative tourniquet duration for the early DVT group [(77.7±12.3) min] was significantly longer than that for the late DVT group [(66.8±11.2) min] and for the DVT-free group [(65.9±10.5) min] ( P<0.05). Conclusions:The majority of postoperative DVTs may be clinically asymptomatic in patients after selective ankle or mid-hindfoot surgery. Although DVT tends to occur within postoperative 2 weeks, its risk may continue after 2 weeks. Increased tourniquet duration may be associated with incidence of early DVT.

Clinical Reasoning Training Learned from Physical Therapy Programs in the USA / 中国康复理论与实践

Clinical reasoning training is the manifestation of comprehensive performance in the students of physical therapy.Strength-ening the clinical-oriented curriculum system,facilitating evidence-based practice knowledge and skills,introducing specialization educa-tion,and applying continuously the evaluation instruments of students'performance in clinical reasoning training would contribute to better education outcome.

Comparative Study Between Complete Ultrasound Guidance and Radiation Guidance in Percutaneous Balloon Pulmonary Valvuloplasty / 中国循环杂志

Objective:To evaluate the safety and efficacy of ultrasound guidance for percutaneous balloon pulmonary valvuloplasty (PBPV) in comparison with conventional X-ray guidance.Methods:Our research included in 2 groups:Ultrasound group,n=102 patients with PBPV under ultrasound guidance in our hospital from 2013-03 to 2016-08 and X-ray group,n=280 patients with PBPV under traditional X-ray guidance in our hospital at the same period of time.Post-operative effect was evaluated by echocardiography and compared between 2 groups.Results:The patients' age,body weight,pulmonary artery diameter,immediate post-operative pulmonary transvalvular pressure gradient (PTPG),the in-hospital time and cost were similar between 2 groups,P＞0.05.The success rate of operation in Ultrasound group and X-ray group was 99.0％ vs 100％,P=0.267.In the ultrasound group,1 patient was converted to a conventional surgery due to right ventricular outflow tract muscle spasm after dilation.The operation time in X-ray group was longer than Ultrasound group,(38.9±9.2) min vs (34.6±10.0) min,P＜0.001.The X-ray exposure time was (3.9±1.2) min in X-ray group.The mean follow-up time was (25.5±13.2) months and PTPG in Ultrasound group and X-ray group were (16.2±4.3) mmHg and (15.3±4.5) mmHg,P=0.120.No serious complications as death,peripheral vascular injury,cardiac perforation and pericardial effusion occurred in either group.Conclusion:PBPV under complete ultrasound guidance may not only avoid radiation and contrast agent,but also keep the safety and efficacy of minimally invasive conventional percutaneous interventional treatment.

Comparative Study Between Complete Ultrasound Guidance and Radiation Guidance in Percutaneous Balloon Pulmonary Valvuloplasty / 中国循环杂志

Objective:To evaluate the safety and efficacy of ultrasound guidance for percutaneous balloon pulmonary valvuloplasty (PBPV) in comparison with conventional X-ray guidance.Methods:Our research included in 2 groups:Ultrasound group,n=102 patients with PBPV under ultrasound guidance in our hospital from 2013-03 to 2016-08 and X-ray group,n=280 patients with PBPV under traditional X-ray guidance in our hospital at the same period of time.Post-operative effect was evaluated by echocardiography and compared between 2 groups.Results:The patients' age,body weight,pulmonary artery diameter,immediate post-operative pulmonary transvalvular pressure gradient (PTPG),the in-hospital time and cost were similar between 2 groups,P＞0.05.The success rate of operation in Ultrasound group and X-ray group was 99.0％ vs 100％,P=0.267.In the ultrasound group,1 patient was converted to a conventional surgery due to right ventricular outflow tract muscle spasm after dilation.The operation time in X-ray group was longer than Ultrasound group,(38.9±9.2) min vs (34.6±10.0) min,P＜0.001.The X-ray exposure time was (3.9±1.2) min in X-ray group.The mean follow-up time was (25.5±13.2) months and PTPG in Ultrasound group and X-ray group were (16.2±4.3) mmHg and (15.3±4.5) mmHg,P=0.120.No serious complications as death,peripheral vascular injury,cardiac perforation and pericardial effusion occurred in either group.Conclusion:PBPV under complete ultrasound guidance may not only avoid radiation and contrast agent,but also keep the safety and efficacy of minimally invasive conventional percutaneous interventional treatment.

An in vitro Vascular Model for Studying the Impact of Different Blood Oxygen Saturation on Main Pulmonary Artery Pressure Changes / 中国循环杂志

Objective: To establish an in vitro vascular model to study the impact of different blood oxygen (O2) saturation on main pulmonary artery pressure changes in order to further explore the mechanism of pulmonary hypertension. Methods: 20 New Zealand white rabbits were randomly divided into 4 groups: Normal control group and 3 experimental groups as High O2 (90％-100％)saturation group, Middle O2 (65％-75％)saturation group and Low O2 (40％-50％) saturation group, n=5 in each group. The in vitro pulmonary artery segments were treated with fresh heparinized rabbit blood and connected to extracorporeal circulation system for 48h at perfusion pressure at 40 mmHg; the condition of O2 saturation was regulated by membrane oxygenator. Pathological morphology was compared among different groups. Results:The bypass time in all 3 groups was 48h and the pressure was stably controlled at 40 mmHg. Blood O2 contents in High O2 saturation group, Middle O2 saturation group and Low O2 saturation group were (97.94±1.01) ％, (72.14±12.85) ％and (43.83±8.71) ％ respectively, P<0.05. Pathological analysis indicated that compared with Normal control group, 3 experimental groups had increased thickness in pulmonary artery wall; upon O2 saturation decreasing, the elastic fibers in pulmonary artery became increasing and more thickening accordingly. Conclusion: We established an in vitro vascular model to study the impact of different blood O2 saturation on main pulmonary artery pressure changes in experimental rabbits.

4-hydroxybenzyl aldehyde can prevent the acute cerebral ischemic injury in rats / 中成药

AIM To investigate the anti-neuroinflammation effects of 4-hydroxybenzyl aldehyde (4-HBAL) from Gastrodia elata Blume on acute cerebral ischemic injury in rats and its nechanism of action.METHODS The rat model of acute cerebral ischemic injury was induced by injecting arachidonic acid via intracarotid artery.Brain tissue samples were taken from the animals 3 h after the model of acute cerebral ischemic injury.Tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) were detected in brain tissue to evaluate the effects of 4-HBAL in vivo.Lipopolysaccharid (LPS)-induced activation of BV-2 microglia cells model was used to explore the anti-neuroinflammation mechanism of 4-HBAL.RESULTS The experimental results showed that 4-HBAL had a significant protective effect on acute cerebral ischemic injury.It could significandy decrease the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β),and obviously inhibit the production of nitric oxide (NO),prostaglandin E2 (PGE2) and TNF-α in LPS-stimulated BV-2 cell,and increase the production of interleukin-10 (IL-10) and transforming growth factors-β (TGF-β) in BV-2 cell.CONCLUSION The mechanism of 4-HBAL may be related to the suppression of the excessive activation of microglia after cerebral ischemia and the promotion of the transformation of microglia into anti-inflammatory phenotype.

Effect of minocycline on activation of microglia M1/M2 phenotypes / 药学学报

This study was designed to investigate the effect of minocycline on microglia activation of M1/M2 phenotypes. The model was induced by lipopolysaccharide (LPS) in BV-2 microglia cells, and was used to evaluate the effect and mechanism of minocycline. We measured nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1 beta (IL-1β) in M1 type microglia, and interleukin-10 (IL-10) and transforming growth factor beta (TGF-β) in M2 type microglia through enzyme linked immunosorbent assay (ELISA). We used flow cytometry to detect the expression of M1 marker CD16/32 and M2 marker CD206 in order to evaluate the influence of minocycline on microglia activation of M1/M2 polarization. Finally, we explored the mechanism of minocycline through detection of the protein expression in response to activation of toll like receptor 4 (TLR4)-mediated myeloid differentiation factor 88 (MyD88) dependent pathway, mitogen activated protein kinase (MAPK) signaling and nuclear factor-κB (NF-κB). The results suggest that minocycline obviously inhibited the production of NO, PGE2, TNF-α and IL-6, and increased the production of IL-10, TGF-β in LPS-stimulated BV-2 cells. Minocycline significantly down-regulated the expression of M1 marker CD16/32 and up-regulated the expression of M2 marker CD206. These results suggest that minocycline can inhibit the activation of microglia to M1 phenotype and promote the transformation of M2 phenotype through down-regulation of p38 and NF-κB signaling pathways.

An in vitro Vascular Model for Studying the Impact of Different Blood Oxygen Saturation on Main Pulmonary Artery Pressure Changes / 中国循环杂志

Objective: To establish an in vitro vascular model to study the impact of different blood oxygen (O2) saturation on main pulmonary artery pressure changes in order to further explore the mechanism of pulmonary hypertension. Methods: 20 New Zealand white rabbits were randomly divided into 4 groups: Normal control group and 3 experimental groups as High O2 (90％-100％)saturation group, Middle O2 (65％-75％)saturation group and Low O2 (40％-50％) saturation group, n=5 in each group. The in vitro pulmonary artery segments were treated with fresh heparinized rabbit blood and connected to extracorporeal circulation system for 48h at perfusion pressure at 40 mmHg; the condition of O2 saturation was regulated by membrane oxygenator. Pathological morphology was compared among different groups. Results:The bypass time in all 3 groups was 48h and the pressure was stably controlled at 40 mmHg. Blood O2 contents in High O2 saturation group, Middle O2 saturation group and Low O2 saturation group were (97.94±1.01) ％, (72.14±12.85) ％and (43.83±8.71) ％ respectively, P<0.05. Pathological analysis indicated that compared with Normal control group, 3 experimental groups had increased thickness in pulmonary artery wall; upon O2 saturation decreasing, the elastic fibers in pulmonary artery became increasing and more thickening accordingly. Conclusion: We established an in vitro vascular model to study the impact of different blood O2 saturation on main pulmonary artery pressure changes in experimental rabbits.

Protective effect of phenylephrine on irradiated injury of osteoblasts in rats / 吉林大学学报(医学版)

Objective To investigate the effect of phenylephrine on the morphology and the expression level of Nampt mRNA in the irradiated osteoblasts of the rats,and to clarify the possible protective effects of phenylephrine on this injury.Methods The osteoblasts cultured by conventional tissue were randomly divided into control group, phenylephrine group,simple irradiation group,and phenylephrine+irradiation group.The cells in phenylephrine group were treated with 100 mmol· L-1 phenylephrine 30 min before irradiation.The cells in simple irradiation group were irradiated with 8 Gy X-ray.The osteoblasts were observed by microscope and were collected at the time points of 8,16,32 and 64 h after irradiation.The Nampt mRNA expression level in osteoblasts was examined by RT-PCR.Results There was no obviously morphological and number changes of osteoblasts in phenylephrine group compared with control group.The osteoblasts in simple irradiation group were atrophied,especially at 8 h;the number of osteoblasts in simple irradiation group was 0.82,0.37,0.24 and 0.21 times as control group respectively at 8,16,32 and 64 h. Atrophy of osteoblasts was alleviated in phenylephrine+irradiation group;at 8, 16,32 and 64 h,the number of osteoblasts was 0.91,0.83,0.72 and 0.75 times as control group,and was 1.09,2.24,3.00 and 3.60 times as simple irradiation group. The expression level of Nampt mRNA in the osteoblasts in simple irradiation group was reduced compared with control group (P < 0.01) at 8, 16, 32 and 64 h.There were significant differernces of the expression levels of Nampt mRNA in the osteoblasts between phenylephrine+irradiation group and control group (P < 0.01),and there were significant differernces between phenylephrine + irradiation group and simple irradiation group (P < 0.01 ) at the different time points. Conclusion Phenylephrine could reduce the atrophy of osteoblasts in the rats induced by irradiation and up-regulate the expression level of Nampt mRNA.Its protective effect on irradiated injury may be related to the up-regulation of the expression level of Nampt mRNA.

Use of Compound Lactobacillus acidophilus Tablets in Rescue Therapy for Failed Eradication of Helicobacter pylori / 胃肠病学

Background:Helicobacter pylori (Hp)infection is correlated with the development of chronic gastritis,peptic ulcer, mucosa-associated lymphoid tissue lymphoma and gastric cancer.Studies showed that Lactobacillus acidophilus could improve the eradication rate of Hp and reduce the occurrence of side-effects.Aims:To evaluate the efficacy of compound Lactobacillus acidophilus tablets combined with bismuth-containing quadruple therapy for the treatment of failed eradiction of Hp infection.Methods:One hundred and eighty patients with Hp-positive peptic ulcer in whom standard triple therapy failed were enrolled and then randomly assigned into experimental group and control group.Patients in experimental group received compound Lactobacillus acidophilus tablets 1 g tid for 14 days,followed by rabeprazole 10 mg bid +amoxicillin 1 000 mg bid +furazolidone 100 mg bid +bismuth 300 mg qid for 10 days.Patients in control group received rabeprazole 10 mg bid +amoxicillin 1 000 mg bid +furazolidone 100 mg bid +bismuth 300 mg qid for 10 days.13 C-urea breath test was conducted at least 4 weeks after completion of the course,Hp eradication rate and side-effects were evaluated. Results:No significant differences in PP eradication rate and ITT eradication rate were found between experimental group and control group (PP:81.2% vs.78.2%,χ2 =0.241,P =0.623;ITT:76.7% vs.75.6%,χ2 =0.031,P =0.861).Incidences of nausea and vomiting (2.4% vs.11.5%),diarrhea (0 vs.9.2%)in experimental group were significantly decreased than those in control group (P <0.05).Conclusions:Compound Lactobacillus acidophilus tablets combined with bismuth-containing quadruple therapy is efficient for Hp-positive patients with failed standard triple therapy and can reduce side-effects.

Recipient dendritic cells modified by RNA interference targeting CD80 and CD86 elicit T cell hyporesponsiveness via enhanced T cell apoptosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Despite extensive research, the mechanism of immature dendritic cells (DCs) induced immune hyporesponsiveness remains incompletely understood.</p><p><b>METHODS</b>Recipient DCs from C3H mouse bone marrow cells were incubated with donor antigen from splenic lymphocytes of C57BL/6 mouse; these DCs were transfected with CD80/86 specific siRNA using lentiviral vectors. Flow cytometry was used to evaluate expression of CD80/86 on the antigen-pulsed recipient DCs. Immune regulatory activity was examined by mixed lymphocyte reaction, in which irradiated DCs were cultured with C3H spleen T cells. After the reaction, interleukin (IL)-2, IL-4, IL-10, and interferon (INF)-γ levels of mixed lymphocyte reaction culture supernatant were measured by enzyme-linked immunosorbent assay. The apoptotic T lymphocytes were identified by Annexin V and CD3 staining.</p><p><b>RESULTS</b>There was a significant inhibition of CD80/86 expression in DCs transfected with CD80/86 lentiviral vectors compared with the control groups (P < 0.05), indicating the specificity of RNA interference. Enzyme-linked immunosorbent assay results showed a significant reduction of INF-γ, IL-2 and IL-10 in the CD80/86 lentivirus transfected group compared to the control groups (P < 0.05). There was no significant difference in IL-4 levels between the groups (P > 0.05). We also showed that CD80/86 low DCs loaded with alloantigen (1) stimulated low T cell proliferative responses via the indirect recognition pathway and (2) enhanced apoptotic activity (P < 0.05) in co-cultured T cells.</p><p><b>CONCLUSIONS</b>Lentiviral vector transfection can effectively and specifically knock down target genes in DCs. The CD80/86 low DCs may show tolerogenic activity via induction of T-cell apoptosis, thereby modulating the activity of recipient-derived DCs. The use of this approach may potentially be clinically applicable.</p>

Development of a yeast two-hybrid screen for selection of A/H1N1 influenza NS1 non-structural protein and human CPSF30 protein interaction inhibitors / 药学学报

Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.

Elucidating the structure of two cyclotides of Viola tianshanica maxim by MALDI TOF/TOF MS analysis / 药学学报

The cyclotides are a family of cyclic "mini" proteins that occur in Violaceae, Rubiaceae and Cucurbitaceae plant families and contain a head-to-tail cyclic backbone and a cystine knot arranged by three disulfide bonds. To study the natural cyclotides of V tianshanica, dried herb was extracted with 50% ethanol, and the concentrated aqueous extract was subjected to a solvent-solvent partitioning between water and hexane, ethyl acetate and n-butanol, separately. The n-butanol extract containing cyclotides was subjected to column chromatography over Sephadex LH-20, eluted with 30% methanol. The subfractions were directly reduced by DTT and analyzed by reverse-phase HPLC. The peaks with different retention times were shown on the profile of RP-HPLC and collected. The cyclotides were speculated based on masses range from 3 000 to 3 500 Da. The purified cyclotides were reduced with DTT, alkylated with iodoacetamide, and then were cleaved with endoproteinase Glu-C, endoproteinase Lys-C and Trypsin, separately. The digested peptides were purified on RP-HPLC and analyzed on MALDI TOF/TOF analyzer. A new cyclotide, cycloviolacin T1 and a reported cyclotide varv E were systemically determined using MALDI TOF/TOF system. So the method for the isolation and characterization of cyclotides was quickly built up in succession.

Elucidating the structure of two cyclotides of Viola tianshanica maxim by MALDI TOF/TOF MS analysis / 药学学报

The cyclotides are a family of cyclic "mini" proteins that occur in Violaceae, Rubiaceae and Cucurbitaceae plant families and contain a head-to-tail cyclic backbone and a cystine knot arranged by three disulfide bonds. To study the natural cyclotides of V tianshanica, dried herb was extracted with 50% ethanol, and the concentrated aqueous extract was subjected to a solvent-solvent partitioning between water and hexane, ethyl acetate and n-butanol, separately. The n-butanol extract containing cyclotides was subjected to column chromatography over Sephadex LH-20, eluted with 30% methanol. The subfractions were directly reduced by DTT and analyzed by reverse-phase HPLC. The peaks with different retention times were shown on the profile of RP-HPLC and collected. The cyclotides were speculated based on masses range from 3 000 to 3 500 Da. The purified cyclotides were reduced with DTT, alkylated with iodoacetamide, and then were cleaved with endoproteinase Glu-C, endoproteinase Lys-C and Trypsin, separately. The digested peptides were purified on RP-HPLC and analyzed on MALDI TOF/TOF analyzer. A new cyclotide, cycloviolacin T1 and a reported cyclotide varv E were systemically determined using MALDI TOF/TOF system. So the method for the isolation and characterization of cyclotides was quickly built up in succession.

Development of a yeast two-hybrid screen for selection of A/H1N1 influenza NS1 non-structural protein and human CPSF30 protein interaction inhibitors / 药学学报

Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.

Effect of total flaveos of Gymostemma pentaphyllum on protein expression of Fas/FasL genes and TNF-alpha concentration in cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of total flaveos of Gymostemma pentaphyllum on the protein expression of apoptosis-associated Fas/FasL gene and tumor necrosis factor-alpha (TNF-alpha) concentration in cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation (H/R).</p><p><b>METHOD</b>A cultured primary neonatal rat cardiomyocytes model with H/R was erected, experiments were divided into six groups, (1)control group, (2)H/R group, (3)15 mg x L(-1) TFG plus H/R group, (4)45 mg x L(-1) TFG plus H/R group, (5) 105 mg x L(-1) TFG plus H/R group, (6)105 mg x L(-1) TFG group. TNF-aconcentration in cultured cardiomyocytes with H/R, was determined by ELISA method, the protein expression of Fas/FasL genes were estimated by immunohisto-chemistry.</p><p><b>RESULT</b>After cardiomyocytes were made with H/R, Compared with control group, the positive expression index (PEI) of Fas/FasL proteins in cardiomyocytes increased significantly, Compared with H/R groups, the PEI of Fas/FasL proteins were lower significantly in groups with different dosages of TFG (P < 0.05). TFG inhibited the secretion of TNF-alpha from myocardial cells and increased the survival rate of myocardial cells.</p><p><b>CONCLUSION</b>The protein expression of apoptosis-associated Fas/FasL genes increased during H/R. The TFG can protect myocardium against H/R injury by decreasing the production of TNF-alpha, downregulating the protein expression of Fas/FasL genes, and then inhibiting myocyte apoptosis.</p>

Expression of alpha1-adrenoceptor and promoting secretory response by phenylephrine in rabbit submandibular gland / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression and distribution of alpha1-adrenoceptor subtypes in rabbit submandibular gland and the effect of phenylephrine on salivary secretion.</p><p><b>METHODS</b>The expressions of alpha1A-, alpha1B-, and alpha1D-adrenoceptor mRNA and protein were investigated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot in rabbit submandibular gland. Immunohistochemical assay was applied to detect the distribution of alpha1A-, alpha1B-, and alpha1D-adrenoceptor and localization of aquaporin 5 in rabbit submandibular gland. Different concentrations of phenylephrine (1 x 10(-8))-(1 x 10(-6)) mol/L were administrated through a polyethylene tube, which was intubated into Wharton's duct of submandibular gland. Heart rate and blood pressure of rabbits were observed during phenylephrine administration. Salivary flow was measured by the length of moist filter paper (35 mm x 5 mm) within 5 min.</p><p><b>RESULTS</b>Alpha1A-, alpha1B-, and alpha1D-adrenoceptor mRNA and protein were expressed in rabbit submandibular gland. Three alpha1-adrenoceptor subtypes were widely distributed in the membrane and cytoplasma of both acinar and ductal cells. Phenylephrine (1 x 10(-7) mol/L, 100 microl) stimulated effectively salivary secretion without inducing significant alteration of blood pressure and heart rate in rabbit. Immunohistochemical assay showed that aquaporin 5 was mainly localized in the apical and lateral plasma membrane in both acinar and ductal cells in unstimulated condition, while the expression of aquaporin 5 was increased after administration of phenylephrine.</p><p><b>CONCLUSIONS</b>Expression of alpha1A-, alpha1B-, and alpha1D-adrenoceptor mRNA and protein was existed in rabbit submandibular gland. Phenylephrine safely and effectively promoted salivary secretion when it was administrated through Wharton's duct of submandibular gland. The mechanism of phenylephrine on salivary secretion may involve in the increase of expression of aquaporin 5 in the apical and lateral plasma membrane in rabbit submandibular gland. This study will hopefully lead to a novel strategy for clinical treatment of dysfunction of submandibular gland.</p>

Analysis on the risk factors associated with fungal infection following operation of gastrointestinal neoplasm / 中国感染与化疗杂志

Objective To investigate the relevant risk factors for fungal infection following operation of the gastrointestinal neo- plasm and offer supporting data for the prevention of fungal infection.Methods Medical records from 116 patients who under- went the operation of gastrointestinal neoplasm in the special group of this hospital from January 2006 to June 2006 were retro- spectively reviewed on the relevant risk factors by univariate and multivariate Logistic regression analysis.Results Of the 116 patients reviewed, 18 had fungal infection.Forty-six samples were positive for fungal pathogen.The most frequently isolated fungal strain was Candida albicans (15/20) and the most common infection site was gastrointestinal tract (14/18).Fungal in- fection after the operation of gastrointestinal neoplasm was significantly relevant with the duration of antibiotic use, duration of post-operative fasting, low serum albumin, high blood glucose and complication of bacterial infection.The duration of antibiotic use was a significantly independent risk factor.Conclusions Reasonable antibiotic use, nutritional support, early enteral nutri- tion and control of blood glucose should be taken into account after the operation of gastrointestinal neoplasm in order to prevent fungal infections.