Volume 91  ·   Issue 10  ·  2011

At present, population aging has become a severe challenge faced by all countries in the world, and it is a strategic problem that affects the long-term economic development<According to the Research Report on Forecasting the Development Trend of China's Population Aging, at the end of 2005, the population over 60 years old in China was nearly 144 million, accounting for 11% of the total population; Aging is not a disease, but it can reduce the threshold of aging-related diseases [1]. Many diseases, such as cardiovascular and cerebrovascular diseases, cancer, etc., are closely related to aging. Therefore, paying attention to the individualized evaluation of aging is of great value to disease prevention and control, especially to "shifting the center of gravity" of cardiovascular disease prevention and treatment, which is helpful to identify the subclinical state of cardiovascular diseases.

Objective To assess the feasibility and safety of laparoscopic rectal surgery without abdominal scar.Methods A total of 23 patients of rectal cancer were operated by laparoscopy with the assistance of PPH (procedure for prolapse and hemorrhoids) anal expander and TEM (transanal endoscopic microsurgery) outer-shell according to the principle of TME (total mesorectal excision).Results All operations were successfully accomplished laparoscopically.There was no conversion into an open procedure.The average operative duration was 129 (105 -211) minutes.The intra-operative blood loss volume was 152(85 -420) ml.No immediate or delayed injury of urinary duct and other intra-operative severe complications,such as massive hemorrhage of presacral venous plexus,occurred.There was no pelvic infection or anastomotic stoma fistula during the post-operative period.The average follow-up period was 13.4 months.Neither anastomotic stoma recurrence nor Trocar implantation occurred.Conclusion The laparoscopic technique of hybrid NOTES (Natural Orifice Transluminal Endoscopic Surgery),plus PPH/TEM is both feasible and convenient for selective rectal cancer surgery.There is no need for extra abdominal incision.It is recommended for laparoscopic rectal surgery.

Objective To study the variation of renal vessels with retroperitoneal laparoscopy so as to increase the safety of retroperitoneal laparoscopic surgeries.Methods A total of 525 patients underwent retroperitoneal laparoscopic nephrectomy or partial nephrectomy at our hospital between January 2004 and June 2008.There were 316 males and 209 females with a mean age of (58 ± 13) years old.The procedures were as follows:(1) patients lay on one side with their waist up and the retroperitoneal cavity was established with our institutional method;(2) gerota's fascia was separated widely along the ventral surface of major psoas muscle;(3) the tissues around renal arteries and veins were isolated by ultrasonic scalpel.Careful observation was performed to explore if there were duplicated or accessory renal vessels;(4) renal vessels were cut by Endo-GIA/Hem-o-lok or blocked by bulldog clamps;(5) whole or partial kidney was finally resected (remaining procedures omitted).Results Among all patients,58 patients (11.0% ,58/525) had a variation of renal vessels.There were double renal arteries on one side (n = 18) ,double renal veins (n = 10),3 renal veins (n = 1) and double arteries and veins on one side (n =3).Twenty-five patients (4.8%,25/525) had one accessory renal artery on one side while 19 (76.0% ,19/25) accessory renal arteries went toward the upper kidney pole.The diameter of one patient's left spermatic vein was similar with that of renal vein and they were joined by lumber vein.Conclusion The variation of renal artery is more common than that of renal vein.The accessory renal arteries are common and usually go toward the upper kidney pole.The variation of renal vessels should be considered before and during a laparoscopic procedure.

Objective To investigate the clinical values of luteinizing hormone-releasing hormone LHRH)α(triptorelin) stimulating test in the differential diagnoses of hypothalamus-pituitary-gonad axis (HPGA) disorders.Methods A total of 229 male patients with various HPGA disorders were recruited for triptorelin stimulating test.And all patients were followed up for 12 -48 months until a definite diagnosis was made.The values of triptorelin stimulating test in the differential diagnoses of HPGA disorders were assessed by examining the close relationship between LHmax and the final clinical diagnosis.Results (1) LH levels rose steady after an intramuscular injection of triptorelin 100 μg and the time of LHmax appeared at 45 -60min.(2) LHmax ＜4 U/L indicated the function of HPGA was not activated.LHmax in the range of 4 - 12U/L indicated the patients might have constitutional delayed puberty development.LHmax ＞ 12 U/Lindicated the fulfilled puberty development.Conclusion Triptorelin stimulating test can precisely evaluate the functions of HPGA in various HPGA disorders and provide valuable information for the differential diagnoses in constitutional delayed puberty development,hypogonadotropic hypogonadolism,central and peripheral precocious puberty disorders.

Objective To discuss the practicability of detecting epidermal growth factor receptor (EGFR) mutations in plasma circulating DNA of lung cancer patients by high-resolution melting (HRM).Methods The sensitivity of HRM was analyzed by the detection of samples containing different proportions of EGFR-mutated plasmids.The mutations in exons 19 and 21 of EGFR were detected by HRM in 96 lung cancer patients from September 2009 to May 2010.And the results of HRM were compared with those of sequencing.Results The HRM detection could identify the EGFR mutations in a proportion of 5% of mutated plasmid DNA.And the EGFR mutations were detected in 17 (17.7%,17/96) cases.Among which,the number of exons 19 and 21 mutations was 15 (88.2%,15/17) and 2 (11.8%,2/17)respectively.The results of sequencing were consistent.Conclusion The HRM analysis may be an optimal method for clinical screening of EGFR mutation due to its simplicity and promptness with a high sensitivity.

Objective To estimate the degree of biological aging for healthy people by the biological age score equation and observe the differences of various aging biomarkers so as to provide targets for clinical anti-aging intervention.Methods A total of 2876 subjects aged 30 - 98 years old were recruited from 3Chinese cities in 2003.After screening,852 healthy subjects were finally selected and assigned into 4 groups according to ages:young group (＜45 yr),middle-aged group (45 -59 yr),young-elder group (60 -74yr) and elder group (≥75 yr).They received a total of 108 physical,morphological,physiologic and biochemical examinations.The biological age score equation was employed to compute the individual biological age scores for all subjects.Then the biological age score was taken as a dependent variable and the chronological age as an independent variable for linear regression.Based on the confidence interval with ± 1standard deviation of regression line,they were divided into 3 groups (delayed aging,normal aging and early aging).According to the chronological ages and degrees of aging,two-way analysis of variance was conducted for the following 7 biomarkers:end diastolic velocity (EDV),intima-media thickness (IMT),ratio of peak velocity of early filling to atrial filling (E/A),mitral valve annulus lateral wall of peak velocity of early filling (MVEL),arterial pulse pressure (PP),fibrinogen (FIB) and cystatin C (CYSC).At the same time,the differences of 7 biomarkers were observed in different aging groups in 4 age groups.Results (1) A comparison of biological age score:there were no significant differences in chronological age among 3biological aging groups in same chronological age groups.However,there were some significant differences in biological age score (young group:F = 91.8,P ＜ 0.01;middle-aged group:F = 134.5,P ＜ 0.01;young-elder group:F = 199.5,P ＜ 0.01;elder group:F = 82.1,P ＜ 0.01).(2) Two-way analysis of variance (aging groups and chronological age groups for biomarkers):there were significant differences of 7biomarkers in different chronological age groups and different aging groups.(3) A comparison of biomarkers among aging groups:there were significant differences in PP among 3 aging groups in 4 age groups.PP increased significantly in early aging group to normal aging group and delayed aging group (young group:49.0±6.9,37.6 ±6.4,30.8±7.6 mm Hg,F=93.2,P＜0.01;middle-aged group:52.9±7.3,44.3 ±5.9,32.7±8.4 mm Hg,F=125.7,P＜0.01;young-elder group:61.9±7.6,51.6±6.6,37.1 ±8.7mm Hg,F=196.5,P＜0.01;elder group:72.2 ±13.7,61.1 ±6.8,43.8 ± 10.8 mm Hg,F =60.2,P ＜0.01).There were significant differences in EDV among 3 aging groups in 4 age groups.EDV increased significantly in early aging group to normal aging group and delayed aging group.There were significant changes in IMT,MVEL,E/A,CYSC and FIB among aging groups in different age groups.Conclusion (1) Biological age score plays an essential role in the evaluation of aging.Based on individual evaluation of biological age score,the degrees of aging can be categorized by grouping so that a clinician may provide clinical anti-aging interventions within the target groups.(2) The above 7 biomarkers are competent for the evaluation of aging.They can not only be used to construct biological age score equation,but also provide clinical targeted interventions for aging.

Objective To investigate the mechanism of azithromycin (AZT) inhibiting the airway mucous hypersecretion through matrix metalloproteinase 9 (MMP9).Methods After culturing,the airway epithelial cells of line HBE16 were stimulated with neutrophil elastase (NE) and pretreated with AZT and epidermal growth factor receptor (EGFR) antagonist BIBX1522.Then the cells were divided into 4 groups:control group,NE-stimulated group,AZT-pretreated & NE-stimulated group and BIBX1522-pretreated & NE-stimulated group.The mucin (MUC)5AC mRNA and MMP9 mRNA levels were analyzed by RT-PCR (reverse transcription-polymerase chain reaction).And the MUC5AC protein content in supernatant was detected by ELISA (enzyme-linked immunosorbent assay).Gelatin zymogrphy was employed to assay the MMP9 activity.Western blot was used to detect the protein levels of MMP9,prozymogen MMP9 (proMMP9),tissue inhibitors of metalloproteinases 1 (TIMP-1),phosphorylated EGFR (p-EGFR) and phosphorylated external-signal regulated kinase (p-ERK).Results As compared with the control group,the levels of MUC5AC and MMP9 gene transcription (0.83 ± 0.17,0.79 ± 0.16) and protein expression (0.84 ±0.15,0.88 ±0.16) in the NE stimulated group were obviously higher than those of the control group (all P ＜ 0.01).And the activity of MMP9 increased (392.33 ± 18.33,P ＜ 0.01) while the protein level of pro-MMP9 decreased (0.17 ±0.10,P ＜0.01).But the expression of TIMP-1 showed no significant change.The protein expressions of p-EGFR and p-ERK increased (0.86 ± 0.23,0.85 ± 0.22,both P ＜0.01);as compared with the NE-stimulated group,there was a reduction of MUC5AC and MMP9 gene transcription (0.36 ± 0.15,0.41 ± 0.09,both P ＜ 0.01) and protein expression levels (0.30 ± 0.08,0.37 ±0.14,both P ＜0.01) in the AZT-pretreated and NE-stimulated group.The MMP9 activity decreased (295.33 ± 14.54,P ＜0.01),the protein levels of pro-MMP9 and TIMP-1 increased (0.46 ±0.14,0.67 ±0.17,both P ＜ 0.05),p-ERK protein level decreased (0.40 ± 0.19,P ＜ 0.01) while the expression of p-EGFR showed no significant decline.As compared with the NE-stimulated group,except for the expressions of MUC5AC mRNA (0.37 ±0.14),MMP9 mRNA (0.37 ±0.13),p-EGFR (0.36 ±0.13) and p-ERK (0.37 ±0.18) decreasing (all P ＜0.01) in BIBX1522-pretreated and NE-stimulated group,the other results had no obvious change.Conclusion AZT may suppress the activity and production of MMP9 in HBE16 cells so as to inhibit the airway mucous hypersecretion.

Objective To express a full-length human-mouse chimeric anti-DR5 antibody from a single open reading frame with tumoricidal activity to various cancer cells.Methods The heavy and light chains of chimeric antibody were joined by the Furin and 2A (F/2A) self-cleavage peptide and cloned into a lentiviral vector of pWPXL.Then the HEK293 cells were infected with the constructed expression vector pWPXL-HF2AL.Western blot,enzyme-linked immunosorbent assay (ELISA) and MTS assay were used to detect the chimeric antibody expression,cleavage,binding affinity to the antigen and tumoricidal activity to various tumor cells.Results The recombinant chimeric antibody was successfully expressed from a single open reading frame in pWPXL-HF2AL construct.And it possessed a similar binding affinity to the parental murine counterpoint and strong tumoricidal activity to various cancer cells.For example,on the concentration of 3 μg/ml,it made the relative cells viability of HCT116,SMMC7721,A549 and U251 down to 20.6% ±2.6% ,35.1% ± 2.7% ,76.1% ±6.1% and 15.6% ±2.0% respectively.Conclusions The human-mouse chimeric anti-DR5 antibody of F/2A peptide is successfully expressed.Possessing a strong tumoricidal activity in various cancer cells,it may provide a novel strategy for cancer biotherapy.

Objective To explore the therapeutic effects of leflunomide metabolite A771726 on high glucose-induced podocyte injury and understand its mechanism.Methods The conditionally immortal human glomeroular podocytes were divided into nomal glucose group (NG) ,high glucose group (HG),mannitol group (MA),high glucose with SB202190 (a p38MAPK inhibitor) group and high glucose with active leflunomide metabolite A771726 group.The levels of p38MAPK and p-p38 protein were determined by Western blot.And the rate of podocyte apoptosis was evaluated by flow cytometry.Results (1) Podocytes with high glucose could activate the p38MAPK signaling pathway.The p-p38 protein expression was significantly elevated.Little activation of the pathways was observed in Groups NG and MA.As compared to HG,the p-p38 protein expression was significantly lowered in SB202190 and A771726 groups.(2) Apoptosis increased in podocytes with high glucose after 24 h.The apoptotic rate of group HG was the most dramatic at 48 h(18.6 ±0.7)%.It was significantly higher than those of Groups NG and MA.The group of high glucose with SB202190 and high glucose with active leflunomide metabolite A771726 could reduce the podocyte apoptosis by 26% and 17% respectively versus the HG group.And the difference was statistically significant.Conclusion Leflunomide can inhibit high glucose-induced podocyte apoptosis.And this effect may be involved in its inhibition of activation of p38MAPK.

Objective To observe the growth and migration of human umbilical cord mesenchymal stem cells (hUCMSCs) on polycarbonate membrane with different pore sizes and explore the criteria of selecting optimal Transwell insert for indirect co-culture to induce the differentiation of hUCMSCs.Methods hUCMSCs were isolated in vitro and then expanded in culture medium.After the treatment of mitomycin C,the cells were seeded on porous membranes of 6-well-dish Transwell inserts with different pore sizes of 0.4,3.0 and 8.0 μm respectively.After culturing for 7 days,the cells were observed and counted on the bottom of each porous membrane.Then the calculation of migration ratio was performed.The growth and migration of hUCMSCs on porous membranes were also examined under scanning electron microscope (SEM).Results The migration ratios of hUCMSCs on membranes of 0.4,3.0 and 8.0 μm pore sizes were 0,1.8% and 8.0% respectively.The migration ratio of cells on 0.4 μm pore size membrane was statistically different from that of the other two pore size groups (P ＜ 0.01).Under SEM,a small portion of cells were growing on the bottoms of membranes and moving through the pores.But there was no cell movement through 0.4 μm pore size membrane.Conclusions hUCMSCs can migrate through the polycarbonate membranes of 3.0 μm and 8.0 μm pore sizes but not through the 0.4 μm one.Thus both sides of polycarbonate membrane of 0.4 μm pore size may be used for close indirect co-culture to induce the differentiation of hUCMSCs.

Objective To explore the influences of rapamycin (RAPA) upon the cytokine changes of activated spleen γδT lymphocytes and lung tissue macrophages in acute lung injury of mice induced by lipopolysaccharide (LPS).Methods A total of 24 healthy male C57BL/6 mice,6 -8 weeks old,were randomly divided into phosphate buffered saline (PBS),LPS,RAPA and LPS + RAPA groups.Acute lung injury was induced by a single intratracheal instillation of LPS in mice.And spleen γδT lymphocytes and lung macrophages were purified by immunomagnetic beads at Day 1.The purified spleen γδT lymphocytes and lung macrophages were adjusted to 106 cell/ml.And 24-well plates were used for 4 groups.Each group were further separated with spleen γδT lymphocytes alone,lung tissue macrophages alone and co-culturing.Supernatant fluid was collected after 24 hours.The expressions of IFN-γ and TNF-o were analyzed by ELISA (enzyme-linked immunosorbent assay).And the expressions of mRNA were analyzed by real-time quantitative PCR (polymerase chain reaction).Results The total cells numbers and lymphocytes numbers of bronchoalveolar lavage fluid were significantly higher in LPS group than those in PBS and LPS + RAPA groups (P＜0.05).And the level of IFN-γwas significantly higher in LPS group than that in PBS,RAPA and LPS + RAPA groups by co-culture (P ＜ 0.05).The level of TNF-α was significantly higher in LPS group than that in RAPA gaud LPS + RAPA groups by co-culture (P ＜ 0.05).However,the mRNA of IFN-γ was higher in LPS group than that in PBS and RAPA groups (P ＜ 0.05).Conclusion RAPA inhibits the secretion levels of IFN-γ and TNF-α in spleen γδT lymphocytes and lung tissue macrophages in acute lung injury of mice induced by LPS.

The patient, a 24-year-old female, went to the Department of Gastroenterology, the First Hospital of Zhejiang University of Traditional Chinese Medicine on November 30, 2009 due to splenomegaly found on physical examination for 12 days. Physical examination showed that the vital signs were stable, the superficial lymph nodes of the whole body were not obviously swollen, the heart and lungs were no different, the abdomen was flat and soft, without tenderness, the liver was 2 cm under the costs, soft and tender, the spleen was about 8 cm under the costs (line A and B was 8 cm, line A and C was 11 cm, line d and E was 0 cm), hard, without tenderness, and the abdominal mass was not palpable. Laboratory examinations: blood routine, urine routine, stool routine, blood biochemistry, thyroid function, erythrocyte sedimentation rate, carcinoembryonic antigen + alpha-fetoprotein + sugar antigen 19-9, complement C3, C4+ immunoglobulin, Faida test, antinuclear antibody, anti-neutrophil cytoplasmic antibody and other laboratory examination indexes were no special abnormalities.

Magnesium is the fourth most abundant cation in human body and the second most abundant cation in cells after potassium. It is a cofactor or agonist of more than 300 enzymes. It participates in many important energy metabolism processes and is closely related to the stability of intracellular environment. In addition to its important physiological effects, magnesium ions also play their own unique advantages in clinical treatment: magnesium sulfate is the first choice for obstetric treatment of eclampsia; The role in the treatment of asthma, ventricular arrhythmia and myocardial ischemia has gradually been affirmed; More and more attention has been paid to the application of magnesium ions in clinical anesthesia. Different routes of administration of magnesium sulfate and its auxiliary analgesic effect and mechanism are briefly described in this paper.

vesicular glutamate transporter (VGLUT) plays an important role in the storage and release of neurotransmitter glutamate, including three subtypes. VGLUT1 and VGLUT2 are the specific markers of all glutamatergic neurons. VGLUT3 is obviously different from the first two subtypes in distribution and function, especially the importance of its relationship with some human diseases has attracted more and more attention in recent years.

A 45-year-old male was admitted to the emergency department of 263 Clinical Department of Beijing Military Region General Hospital in September 2010 mainly due to persistent retrosternal dull pain for 4 h. He denied history of hypertension and diabetes. Physical examination: blood pressure 120/80 mm Hg (1 mm Hg =0.133 kPa), clear respiratory sounds in both lungs, no dry and wet rales, heart rate 102 beats/min, harmonic heart rhythm, low and dull heart sounds, no pathological murmur, and no swelling in both lower limbs. Laboratory examination: blood urea nitrogen 7.98 mmol/L, blood creatinine 62 mmol/L, blood potassium 4.2 mmol/L, alanine aminotransferase (ALT) 50 U/L, aspartate aminotransferase (AST) 56 U/L, brain natriuretic peptide 16.3 ng/L, troponin 0.25 μ g/L, blood glucose 6.42 mmol/L. ECG: acute anterior wall myocardial infarction.

Nearly half of the 33.4 million people infected with human immunodeficiency virus (HIV) in the world are women of childbearing age, and almost all of the 2.1 million infected children are transmitted during pregnancy, childbirth or breastfeeding. Since 2002, the number of newly infected children has decreased gradually, probably due to the implementation of mother-to-child HIV blockade.