The roles of sphingosine-1-phosphate and its receptor in Mycobacterium tuberculosis infection and novel antibiotics discovery / 药学学报

Tuberculosis (TB) caused by Mycobacterium tuberculosis (M. tuberculosis) infection remains a major public health problem of global concern, largely due to antibiotics resistance, persistence and immune evasion. Sphingolipid bioactive molecules are involved in several important pathophysiological processes. Sphingosine-1-phosphate is a key product of sphingolipid metabolism, and can play a role in two manners: autocrine and/or paracrine. Sphingosine-1-phosphate regulates T cells and a variety of antigen-presenting cells during M. tuberculosis infection, promotes antigen processing and expression in monocytes, is involved in the maturation of phagolysosome, regulates Ca2+ homeostasis, participates in the autophagy of macrophages, inhibits the survival and proliferation of M. tuberculosis within host cells, and effectively reduces the necrosis of the mouse lungs infected by M. tuberculosis. Injection of 20 nmol per mouse sphingosine-1-phosphate inhibited up to 47% of mycobacterial growth in the lung and spleen of mice infected by M. tuberculosis. In this paper, sphingosine-1-phosphate, its receptors and regulatory network were reviewed, and the specific mechanism of sphingosine-1-phosphate inhibiting the survival of M. tuberculosis-infected host cells was elaborated. This will provide novel insights into the new targets for tuberculosis prevention and treatment.

Effects of Curcumin on in vitro and in vivo Proliferation and Invasion of Osteosarcoma Cells and Its Mechanism Study / 中国药房

OBJECTIVE:To investigate the effects of curcumin on proliferation and invasion of osteosarcoma cells and its mechanism. METHODS:In in vitro cell assay,osteosarcoma cells were divided into control group(dimethyl sulfoxide),curcumin low-concentration,medium-concentration and high-concentration groups(10,20,40 μmol/L)and curcumin high-concentration+endogenous miR-21 simulacrum(agomiR-21)group,curcumin high-concentration+agomiR-21 negative control(NC)group. The proliferation(OD value)and invasion ability(transmembrane cells number)of human osteosarcoma cells U-2OS and MG-63 were detected by CCK-8 and Transwell chamber method;the expression of miR-21 and its target gene PDCD4 protein were detected by real-time fluorescence quantitative PCR and Western blot assay. In in vivo cell assay,MG-63 cells were inoculated in mice,and the mice were divided into blank control group(normal saline),curcumin group(20 mg/kg),curcumin+agomiR-21 NC group(20 mg/kg curcumin solution+50 mg/kg agomiR-21 NC solution),curcumin+agomiR-21 group(20 mg/kg curcumin solution+50 mg/kg agomiR-21 solution)with 8 mice in each group;they were given intratumoral injection of relevant medicine once a day,for consecutive 7 d. The volume changes of the tumor were measured within 20 d after medication,and the expression of miR-21 and PDCD4 protein in tumor tissue were detected on 20th day. RESULTS:Compared with control group,the OD value and transmembrane cells number of U-2OS and MG-63 cells were decreased significantly in curcumin low-concentration, medium-concentration and high-concentration groups(P<0.05 or P<0.01);the expression of miR-21 was weakened significantly in curcumin medium-concentration and high-concentration groups of U-2OS and MG-63 cells(P<0.05 or P<0.01),while the expression of PDCD4 protein was strengthened significantly(P<0.01). Compared with curcumin high-concentration+agomiR-21 NC group,the OD value and transmembrane cells number of U-2OS and MG-63 cells were strengthened significantly in curcumin high-concentration+agomiR-21 group(P<0.01). Compared with blank control group,the tumor volume of mice were decreased significantly in curcumin group and curcumin+agomiR-21 NC group(P<0.05 or P<0.01). The expression of miR-21 was decreased significantly in tumor tissue(P<0.01),while the protein expression of PDCD4 was strengthened significantly(P<0.01). Compared with curcumin+agomiR-21 NC group,the tumor volume of mice were increased significantly in curcumin+agomiR-21 group(P<0.05 or P<0.01). The expression of miR-21 in tumor tissue were strengthened significantly(P<0.01),while the expression of PDCD4 protein were weakened significantly(P<0.01). CONCLUSIONS:Curcumin can inhibit the proliferation and invasion of human osteosarcoma cells U-2OS and MG-63,the mechanism of which may be associated with down-regulating the expression of miR-21.

Clinical observation on acupoint pressure therapy plus electroacupuncture for post-traumatic knee osteoarthritis / 针灸推拿医学（英文版）

Objective:To investigate the clinical efficacy of Zhang's acupoint pressure therapy plus electroacupuncture (EA) in treating post-traumatic knee osteoarthritis.Methods:A total of 98 eligible patients with post-traumatic knee osteoarthritis were divided into group A and B by the random number table, 49 cases in each group. Group A was intervened by Zhang's acupoint pressure therapy plus EA; group B was given medicinal fumigation. The clinical efficacies of the two groups were compared.Results:The markedly effective rate of group A was significantly higher than that of group B.Conclusion:Zhang's acupoint pressure therapy plus EA can produce a satisfactory clinical efficacy in treating post-traumatic knee osteoarthritis, and is worth promotion.

Research progress in autophagy on dry age-related macular degeneration / 中华实验眼科杂志

Age-related macular degeneration (AMD) is a common cause of vision loss and living quality impairment in elderly people.Dry AMD is considered to be a neurodegenerative disease, and there has been no preventive method or effective therapy for it so far.Recent studies reveal that accumulation of lipofuscin may lead to dysfunction and even death of retinal pigment epithelium (RPE) cells.Autophagy is a major lysosome-dependent degradation pathway in eukaryotes involved in the disposal of damaged cytoplasmic proteins and organelles.Autophagy is revealed to be involved in the pathological processes of several neurodegenerative diseases such as Alzheimer's disease and dry AMD.Therefore,studies focus on autophagy may provide a new target for the prevention and treatment of dry AMD.This paper reviewed the research progress of autophagy in the pathogenesis of AMD in recent years.The roles of autophagy,lysosomal damage,oxidative stress and immune inflammatory reaction were described.

The study of catheterization to measure ventricular pressure in mice through venous cannula in external jugular vein / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study a feasible method of measuring right ventricular pressure by catheterization in mice.</p><p><b>METHODS</b>Measuring the right ventricular pressure and the pulmonary artery pressure by homemade PE pipe through venous cannula in external jugular vein, using catheterization in mice with powerlab multimodal biometric signal recording system.</p><p><b>RESULTS</b>Forty-six out of 51 mice were experimented with this method smoothly and got a total success rate of 90.2%. Thirty of 33 normal mice and 16 of 18 mice with pulmonary arterial hypertension (PAH) were catheterized successfully. The right ventricular pressure were as follow: systolic blood pressure: (23.4 +/- 5.7) mmHg in normal group vs (32.2 +/- 2.8) mmHg in mice with PAH, diastolic blood pressure: (3.7 +/- 2.6) mmHg vs (3.8 +/- 2.0) mmHg, mean pressure: (12.0 +/- 3.7) mmHg vs (14.9 +/- 2.3) mmHg. After autopsy for those 5 failed cases, we found that 2 cases were into the inferior vena cava, another 2 cases pierced the right auricle and the last one punctured the axillary vein into the chest wall.</p><p><b>CONCLUSION</b>Measuring the right ventricular pressure through venous cannula in external jugular vein with homemade PE pipe in mice gets not only a high success rate but also help to save time. Moreover, this method can be popularized easily. It is a good and feasible method for measuring right ventricular pressure in mice.</p>

The Stent-Assisted Coil-Jailing Technique Facilitates Efficient Embolization of Tiny Cerebral Aneurysms

OBJECTIVE: Tiny cerebral aneurysms are difficult to embolize because the aneurysm's sac is too small for a single small coil, and coils within the aneurysm may escape from the confinement of a stent. This study was performed to introduce the stent-assisted coil-jailing technique and to investigate its effect on the coil embolization of tiny intracranial aneurysms. MATERIALS AND METHODS: Sixteen patients with tiny intracranial aneurysms treated with the stent-assisted coil-jailing technique between January 2011 and December 2013 were retrospectively reviewed and followed-up. RESULTS: All aneurysms were successfully treated with the coil-jailing technique, and at the end of embolization, complete occlusion of the aneurysm was achieved in 9 cases (56.3%), incomplete occlusion in 6 (37.5%), and partial occlusion in 1 (6.3%). Intraprocedural complications included acute thrombosis in one case (6.3%) and re-rupture in another (6.3%). Both complications were managed appropriately with no sequela. Follow-up was performed in all patients for 3-24 months (mean, 7.7 months) after embolization. Complete occlusion was sustained in the 9 aneurysms with initial complete occlusion, progressive thrombosis to complete occlusion occurred in the 6 aneurysms with initial near-complete occlusion, and one aneurysm resulted in progressive thrombosis to complete occlusion after initial partial occlusion. No migration of stents or coils occurred at follow-up as compared with their positions immediately after embolization. At follow-up, all patients had recovered with no sequela. CONCLUSION: The stent-assisted coil-jailing technique can be an efficient approach for tiny intracranial aneurysms, even though no definite conclusion regarding its safety can be drawn from the current data.

The efficiency and safety of laparoscopic surgery for colorectal carcinoma: a systematic review / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate and compare the efficiency and safety of laparoscopic surgery (LS) and open surgery (OS) in the treatment of colorectal carcinoma.</p><p><b>METHODS</b>Randomized controlled trials on laparoscopic surgery and open surgery for colorectal carcinoma from January 2000 to October 2010 were searched in the databases of EMbase, PubMed, Cochrane Library, Sciencedirect, Springer, VIP, CNKI, CBMdisc. The methodological quality was assessed according to the standard of Cochrane systematic review. For homogeneous studies, RevMan5.0 software was used for meta-analysis.</p><p><b>RESULTS</b>A total of 13 RCTs involving 4603 patients were included in this study, and among those 6 were multi-center randomized controlled trials. The meta-analysis showed that: the operation time of the LS group was longer than that of the OS group (WMD = 38.91, 95%CI: 33.89 - 43.93, P < 0.001), the blood loss (WMD = -138.14, 95%CI: -195.79 - -80.50, P < 0.001) and the length of hospital stay (WMD = 2.91, 95%CI: -4.65 - -1.17, P = 0.001) of the LS group was less than those in OS group. There was no significant differences between the two groups in the number of dissected lymph nodes (WMD = -0.62, 95%CI: -1.47 - 0.23, P = 0.150). There was no significant differences between the two groups in terms of the postoperative complications (30 days) (RR = 0.78, 95%CI: 0.59 - 1.01, P = 0.06). There was no significant differences between the two groups in 3-year overall survival (RR = 1.00, 95%CI: 0.96 - 1.04, P = 0.970). There was no significant differences between the two groups in 5-year overall survival (RR = 1.03, 95%CI: 0.99 - 1.08, P = 0.140). There was no significant differences between the two groups in 5-year overall recurrence (RR = 0.89, 95%CI: 0.74 - 1.07, P = 0.200).</p><p><b>CONCLUSIONS</b>Laparoscopic surgery for colorectal carcinoma is a safe and effective therapy as open surgery in the short term or long term outcomes. It could be an acceptable alternative to open surgery for colorectal carcinoma.</p>

The ectopic expression and quantitative analysis of Integrin beta1 during wound healing process after mixed microskin transplantation / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the mechanism of promoting wound healing with mixed grafting of autologous and allogeneic microskin in rats.</p><p><b>METHODS</b>Fifteen male Wistar rats served as alloskin donor. Forty-five female SD rats with full-thickness skin defect served as recipients in the study. In part one experiment, 27 SD rats were randomly divided into group I (n = 9, without allogeneic microskin), group II (n = 9, with mixed grafting of allogeneic microskin at area expansion rate of 10:1); group III (n = 9, with mixed grafting of allogeneic microskin at area expansion rate of 10:3) with grafting with the same amount of autologous microskin at area expansion rate of 10:1. In part two experiment, 18 SD rats were also divided into group I (n = 6, with autologous microskin only); group II (n = 6, with mixed grafting of autologous and allogeneic microskin with area expansion rate of 20:1 and 20:3 respectively); group III (n = 6, with mixed grafting of autologous and allogeneic microskin with area expansion rate of 20:1 and 20:6, respectively). Biopsy samples were obtained from healed wound area of SD rats in each group at different time points after operation. The histological changes, epidermal thickness, and immunohistochemical staining of Integrin beta1 were observed.</p><p><b>RESULTS</b>(1) HE staining showed the thickness of epidermis in each group increased obviously, and various amounts of mononuclear cell infiltration and different degrees of vasodilation appeared in the dermal layer during 2 - 4 weeks. (2) Epidermal thickness in group II and III of part one experiment were significantly thicker than that in group I during 2 - 4 weeks after operation (P < 0.05), and the similar result was also seen in part two experiment on 3 and 4 weeks after operation (P < 0.05). (3) A positive staining pattern for Integrin beta1 was seen in the suprabasal layers (especially in the spinous and granular layers) in all groups. In part one experiment, the expression of Integrin beta1 in group II and III were obviously higher than that in group I on 2 week after operation (P < 0.01), and the expression of Integrin beta1 in group II (10 982 +/- 2169) was also higher than that in group III (4240 +/- 512, P < 0.01); the expression of Integrin beta1 in group II was still higher than that in group I and III (P < 0.01) 3 and 4 weeks after operation. In part two experiment, the expression of Integrin beta1 in group III (1618 +/- 171) was higher than that in group I 3 weeks after operation (1060 +/- 146, P < 0.05).</p><p><b>CONCLUSION</b>The ectopic and increased expression of Integrin beta1 was closely associated with the proliferation and differentiation of epidermal cells, wound reepithelialization and thickened epidermis in mixed grafting of autologous and allogeneic microskin. Integrin beta1 may be responsible in promoting wound healing.</p>

Study on the optimal ratio of autoskin and alloskin during mixed microskin grafting / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the influence of different amount of allogeneic microskin in mixed grafting with certain quantity of autologous microskin on wound healing in rats. Methods Male Wistar rats served as alloskin donor rats. Forty female SD rats with full thickness skin defect were enrolled in the study, and they were randomly divided into four groups, i.e. group I (n=10, with allogeneic microskin graft at area expansion rate of 10:3); group II (n=10, with autologous microskin graft at area expansion rate of 10:1); group III (n=10, with mixed grafting of autologous and allogeneic microskin at area expansion rate of 10:1, respectively); group IV (n=10, with mixed grafting of autologous and allogeneic microskin at area expansion rate of 10:1 and 10:3, respectively). The wound healing rate, wound contraction rate and histological changes were observed at the 2, 3 and 4 post graft weeks (PGW).</p><p><b>RESULTS</b>(1) In group I, there was mainly granulation tissue with some de novo epithelial cells appearing at the wound edge along with the rejection of grafted allogenous skin in the rat wound. In group II, there was still some granulation tissue remaining at 2 PGW due to insufficient amount of microskin. However, the wounds in the mixed grafting group appeared almost totally epithelialized. (2) Various amounts of mononuclear inflammatory cell infiltration and different degrees of angiectasis were observed in the dermal layer after the skin grafting in all groups, especially in group II and IV. There was thickening of the epithelial layer in all groups except group I. (3) The wound healing rate decreased obviously along with the development of rejection in group I at 2 to 4 PGWs. The wound healing rate was (55 +/- 26)% in group II, which was obviously lower than that in group III (88 +/- 6)% and in group IV (76 +/- 10)% at 3 PGWs (P < 0.01). (4) The contraction rate of the wound in group IV (69 +/- 7)% was much higher than that in group I (58 +/- 11)% at 3 PGWs (P < 0.05), and there was no difference among all the other groups.</p><p><b>CONCLUSION</b>Wound healing can be obviously accelerated by mixing some autologous microskin with appropriate amount of alloskin. Moreover, certain amount of autologous microskin (expansion rate 10:1) mixed with the same proportion of allogeneic microskin seems to be more beneficial in promoting wound healing.</p>

Prolongation of the survival of skin allografts by intravenous injection of neuraminidase-treated donor bone marrow cells in rats / 中华烧伤杂志

<p><b>OBJECTIVE</b>To optimize the best concentration of neuraminidase (Neu) that enhances the migration of neuraminidase (Neu)-treated donor bone marrow cells (dBMCs) to the liver, and observe the influence of short-term cyclosporin A(CsA) application combined with intravenous injection (i.v.) of Ne treated dBMCs on the survival of skin allografts.</p><p><b>METHODS</b>The experiment consisted of two parts. For selection of an appropriate concentration of Neu, 26 female Wistar rats were randomly divided into four groups. The dBMCs were prepared by routine method and treated with four concentrations (0, 0.5, 1.0, 2.0 U/ml) of Neu at 37 degrees C for 30 min. The untreated and Neu-treated dBMCs were labeled by 99mTc, and injected via the tail veins to female Wistar rats in each group, respectively. After five hours, the radioactivity of various organs collected from sacrificed rats was measured by a gamma counter, and the values were expressed as percentage of total radioactivity of all organs from the same rat. To observe the survival of skin allograft, 23 male Wistar rats were randomly divided into control group, untreated dBMCs group and Neu-treated dBMCs group. All rats in each group were grafted with skin allografts from male Sprague-Dawley (SD) rats. The dBMCs from the same donor without and with Neu treatment by the concentration selected from the above experiment were injected via the tail veins of female Wistar rats in untreated dBMCs group and Neu-treated dBMCs group, respectively. Rats in untreated dBMCs group and Neu-treated dBMCs group received CsA (10 mg/kg) through intraperitoneal injection (i.p.) at 2 and 5 days post-grafting. Neither dBMCs or CsA were given in the control group. The survival of allograft skin in each group was checked and photographed daily after 5 days post operation.</p><p><b>RESULTS</b>When the concentration of Neu was 1.0 U/ml, the percentage of dBMCs in liver was (75.3 +/- 9.8) %, which was obviously higher than that in 0 U/ml group [(58.9 +/- 4.2%)], (P < 0.01), indicating that the optimal concentration of Neu was 1.0 U/ml. The survival time of skin allografts in rats of Neu-treated dBMCs group was prolonged significantly in comparison with that of the rats in dBMCs group without Neu treatment (P < 0.01). The survival time in both dBMCs group and Neu-treated dBMCs group was longer that of control group (P < 0.01), and it was prolonged in Neu-treated dBMCs group compared with that in dBMC group.</p><p><b>CONCLUSION</b>Administration of proper concentration of Neu can increase the affinity of dBMCs to the liver, and promote the Neu-treated dBMCs to migrate to liver. The intravenous injection of Neu-treated dBMCs combined with short-term CsA administration can delay the rejection of skin allografts in rats.</p>

RNA interference-mediated silencing of MAT 2A gene attenuates growth and induces apoptosis of hepatoma cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effect of short interfering RNA targeting MAT 2A on growth and apoptosis of hepatoma cells.</p><p><b>METHODS</b>The four siRNA against MAT 2A gene were transcript synthesized intracelluarly by expressed templates of plasmid vector pSilence-2.1-U6. We inserted the target sequence of MAT 2A gene into the upstream of the reporter gene in order to construct the recombinant plasmid vector plucA-MAT 2A. The recombinant plasmid and siRNA-producing plasmid were co-transfected into 293 T cells using this construct via lipofectamine methods. The inhibition effect was detected by measuring luciferase activity in the cell lysate to screen the effective siRNA, and then, the effective siRNA was transfected into Bel-7402 cells. The effect of siRNA treatment on the MAT 2A mRNA level and the MAT activity of hepatoma cells were measured. In order to study the effect of short interfering RNA targeting MAT 2A on growth and apoptosis of hepatoma cells, the tumor cell killing rate was analyzed by MTT method and the rate of apoptosis of hepatoma cells was evaluated by flow cytometry.</p><p><b>RESULTS</b>The two siRNA among the four siRNA displayed inhibitory effect on the lucifermase expression with the inhibitory rates of 81% and 89% respectively. The expression of MAT 2A mRNA in Bel-7402 cells was specifically inhibited and the MAT activity in Bel-7402 cells was decreased. Furthermore, silencing of the MAT 2A gene by RNAi significantly inhibited hepatoma cell growth and led to induction of apoptosis.</p><p><b>CONCLUSION</b>RNA interference-mediated silencing of MAT 2A gene attenuates growth and induces apoptosis of hepatoma cells; MAT 2A is an ideal target of gene-specific therapy for liver cancer.</p>

The treatment choice of solitary kidney complicated with complex calculi report of 42 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the therapeutics of complex calculi in solitary kidney, and to improve the effect and safety of treatment.</p><p><b>METHODS</b>Experiences in the treatment of 42 patients were summarized. All patients were with mould or multiple calculi, 8 cases were complicated with ureter calculi, and 6 cases were hospitalized because of obstructive anuria. The patients with mould calculi received extracorporal shock wave lithotripsy (ESWL) prior to percutaneous nephrolithotomy (PCNL). While the patients with multiple calculi received PCNL prior to ESWL. Some cases were treated by lithotripsy.</p><p><b>RESULTS</b>Thirty-six cases (86%) were cured by ESWL combined with PCNL. Eleven cases received lithotripsy during PCNL. The 6 cases with obstructive anuria recovered in 12 hours after emergent ESWL or lithotripsy; 6 cases (14%) underwent open operation because of deformity or obstruction in renal pelvis and ureter; 2 cases have to keep nephrostomy because of repeated infection. Followed up 6-18 months, 38 cases (86%) keep good kidney function; 5 cases (14%) had renal insufficiency; 4 cases (11%) reoccurred calculi.</p><p><b>CONCLUSIONS</b>The therapeutics of ESWL combined with PCNL may clear complex calculi of solitary kidney effectively and safe. It is necessary to take emergent ESWL in renal obstructive calculi cases. And the patients with lower ureter obstructive calculi may take lithotripsy first. It is proper to choose open operation on the patients with deformity of renal pelvis or obstruction of ureter.</p>

Cerebral Proerectile Effect of Apomorphine and Vasoactive Agents in Anesthetized Rats / 대한비뇨기과학회지

PURPOSE: To establish central control of a penile erection, the centrally elicited erectile effects of apomorphine hydrochloride and various vasoactive agents were investigated after intracerebroventricular administration. MATERIALS AND METHODS: Anesthetized male Sprague-Dawley rats were used for the study. After intracerebroventricular administration of normal saline(NS), apomorphine hydrochloride(AH), prostaglandin E1(PGE1), sodium nitroprusside(SNP), phenylephrine hydrochloride(PE) and phentolamine(PT) under a stereotaxic setting, the intracavernosal pressure(ICP), systolic femoral artery pressure(FAP), heart rate(HR), time to first response, duration and number of erectile response and adverse reactions were evaluated for 60 minutes. To show if the centrally elicited erection by agents in native setting were effective, the above criteria were re-evaluated after a bilateral pelvic neurotomy and bilateral orchiectomy. RESULTS:Cerebral proerectile effects were elicited by AH with no significant changes in the FAP and HR, but with PGE1, SNP, PE and PT significant changes in both the FAP and HR were observed. The ICP/FAP ratio was highest with SNP at 0.75+/-0.08. The mean time to first response was shortest with AH at 18.1+/-5.1min. The mean duration was longest with AH at 39.4+/-10.9min. The number of responses was highest with AH at 2.7+/-1.1. Adverse reactions, such as stretching, yawning and ejaculation, were simultaneously observed during increases in the ICP. In the case of a bilateral pelvic neurotomy or bilateral orchiectomy, these elicited erectile responses disappeared. CONCLUSIONS: Vasoactive agents, such as SNP, PGE1 and PE, had a cerebral proerectile effect, but AH showed more an evidently potent proerectile effect in the aspects of ICP, time to first response and duration of erectile response. Testosterone and the pelvic nerve are suggested to be essential for a central proerectile response.

Construction and sequencing of recombinant lentiviral plasmid affecting human CCL20 gene by RNA interfering / 第三军医大学学报

Objective To design the small interference RNA (siRNA) specific to human CCL20 gene by RNA interfering technique, construct its recombinant lentiviral expression vectors, and identify these vectors by DNA sequencing. Methods According to Tuschl’s principle, the siRNA was designed and converted into cDNA of shRNA (small hairpin RNA) of siRNA for CCL20 gene. The cDNA was synthesized and inserted into plasmid pHSER-dsRNA-GFP-SIN which was linearized by restriction endonucleases SpeⅠ and SalⅠ. The recombinant plasmid was transformed into competent E. coli. DH5? cells. The positive recombinant colony was selected by ampicillin medium agar and identified by DNA sequencing. Results Two recombinant lentiviral plasmids of siRNA specific to CCL20 gene were constructed successfully. Their DNA sequence analysis completely coincided with their designed sequences. Conclusion Lentiviral vector-based siRNA expression plasmids against CCL20 gene have been successfully constructed and identified. They will be further used for interfering CCL20 mRNA transcription and lay the foundation for CCL20 gene modified human keratinocyte stem cells.

Surgical Outcome of 153 Vasovasostomies on 10 Years or More after Vasectomy / 대한비뇨기과학회지

PURPOSE: In the 1970s, male sterilization (vasectomy) has been performed on a large scale as an accepted method of family planning in Korea. Since then, especially during last decade, there has been an increase in the number of patients requesting vasovasostomy after a long term interval following a vasectomy. MATERIALS AND METHODS: We evaluated 153 cases of vasovasostomy performed, 10 years or more after a vasectomy, at the Pusan National University Hospital between March 1983 and March 2002. Clinical data were collected through telephone interviews and a survey of medical records. RESULTS: The ages at the time of reversal ranged from 30 to 57, with an average of 41.6 years. The obstructive intervals ranged from 10 to 24, with an average of 17.0 years. To the cases were divided into 3 the obstructive intervals; 10 to 14, 15 to 19 and 20 to 24 years, with 130 (85.0%), 15 (9.8%) and 8 cases (5.2%), respectively. The overall patency and pregnancy rates were 81.6 and 36.0%, respectively. There were no significant differences according to the methods of vasovasostomy, suture materials, ooze from the proximal vasal end, the presence of sperm granuloma, the levels of anastomosis or the age of patients, with the exception of the ages of the partners in the patency and pregnancy rates for the patients with vasovasostomy 10 years or more after vasectomy. CONCLUSIONS: Our results suggest that the pregnancy rates after vasovasostomies, following at least a 10 years long term obstruction, are very low, despite the desirable patency rates. Therefore, it is important that we should apply the assisted reproductive technology, as well as advanced surgical skills, to improve the pregnancy rates.

The Antioxidant Effect of Rebamipide in Human Semen Specimens / 대한비뇨기과학회지

PURPOSE: Rebamipide is a propionic acid derivative that inhibits superoxide production and removes hydroxyl radicals. This study was performed to investigate the effects of adding rebamipide to semen, in an effort to determine if reactive oxygen species (ROS) production and lipid peroxidation of the sperm cell membrane as well as an improvement in seminal parameter and fertilizing capacity under oxidative stress was inhibited. MATERIALS AND MTHODS: Semen was collected from 30 normal healthy volunteers by masturbation after at least 48 hours abstinence. After liquefaction of the semen at room temperature, the prepared sperm was diluted with a sperm wash media to a uniform density of 20x106/ml. The semen was treated with 0.25ml of 0.2mM FeSO4 and 1mM sodium ascorbate for 60 min in the presence of various rebamipide concentrations (0, 10, 30, 100, and 300microM). ROS production, sperm motility, vitality, fertilizing capacity and the level of lipid peroxidation were analyzed by chemiluminescence, computer assisted semen analysis, eosin-nigrosin staining, a hypo-osmotic swelling test and the thiobarbituric acid method, respectively. RESULTS: Rebamipide at 100 and 300microM increased the sperm motility (p<0.05) but did not affect the sperm vitality. The ROS production and lipid peroxidation in the sperms treated with FeSO4/sodium ascorbate were inhibited by rebamipide in a dose-dependent fashion (p<0.05 in each). The total swelling rate of the hypo-osmotic swelling test was also increased by high rebamipide concentrations (100 and 300microM), respectively 49.2 17.9 and 50.8 21.7% (p<0.05). CONCLUSIONS: These results suggest rebamipide is an effective free radical scavenger and may be useful as an oral antioxidant in patients with male infertility due to increased ROS generation. However, further study to be possible the clinical use of rebamipide for improve the fertilizing capacity in male infertility is required.

The Antioxidant Effect of Rebamipide in Human Semen Specimens / 대한비뇨기과학회지

PURPOSE: Rebamipide is a propionic acid derivative that inhibits superoxide production and removes hydroxyl radicals. This study was performed to investigate the effects of adding rebamipide to semen, in an effort to determine if reactive oxygen species (ROS) production and lipid peroxidation of the sperm cell membrane as well as an improvement in seminal parameter and fertilizing capacity under oxidative stress was inhibited. MATERIALS AND MTHODS: Semen was collected from 30 normal healthy volunteers by masturbation after at least 48 hours abstinence. After liquefaction of the semen at room temperature, the prepared sperm was diluted with a sperm wash media to a uniform density of 20x106/ml. The semen was treated with 0.25ml of 0.2mM FeSO4 and 1mM sodium ascorbate for 60 min in the presence of various rebamipide concentrations (0, 10, 30, 100, and 300microM). ROS production, sperm motility, vitality, fertilizing capacity and the level of lipid peroxidation were analyzed by chemiluminescence, computer assisted semen analysis, eosin-nigrosin staining, a hypo-osmotic swelling test and the thiobarbituric acid method, respectively. RESULTS: Rebamipide at 100 and 300microM increased the sperm motility (p<0.05) but did not affect the sperm vitality. The ROS production and lipid peroxidation in the sperms treated with FeSO4/sodium ascorbate were inhibited by rebamipide in a dose-dependent fashion (p<0.05 in each). The total swelling rate of the hypo-osmotic swelling test was also increased by high rebamipide concentrations (100 and 300microM), respectively 49.2 17.9 and 50.8 21.7% (p<0.05). CONCLUSIONS: These results suggest rebamipide is an effective free radical scavenger and may be useful as an oral antioxidant in patients with male infertility due to increased ROS generation. However, further study to be possible the clinical use of rebamipide for improve the fertilizing capacity in male infertility is required.

Oral Testosterone Decanoate (Andriol(R)) Supplement Therapy Improve the Quality of Life in the Men with Testosterone Deficiency / 대한남성과학회지

PURPOSE: We evaluated the changes in quality of life after administering an oral testosterone supplement to men with testosterone deficiencies. MATERIALS and METHODS: The changes of subjective symptoms were estimated in 39 cases (33 cases in the experimental group and 6 cases in the placebo group) of testosterone definiency,in men who were treated with oral testosterone undecanoate (Andriol(R)) or a placebo by the single blind method from March 2001 to January 2002. The 80mg dose of oral testosterone was administered twice daily in a period of three months. Digital rectal examinations and laboratory tests including serum testosterone, PSA, cholesterol, AST and ALT were conducted on pretreatment and during the 1st and 3rd months after treatment. The changes in quality of life were evaluated on pretreatment and monthly after treatment by the PNUH QOL Scoring System, which consisted of 21 question sinquiring about the following 7 functions: metabolic, cardiopulmonary, musculoskeletal, gastroinestinal, neurologic and psychiatric and sexual. The Saint Louis University Questionnaire was used simultaneously to evaluate the changes in quality of life. RESULTS: The 39 cases showed a 19~64 years old age distribution (mean 45.9 years old). Underlying causes of testosterone deficiency were primary hypogonadism and andropause in 10 and 29 cases, respectively. During the follow-up period, the drop-out after treatment in the experimental group was 4 cases due to low compliance or personal reasons. Serum testosterone was significantly increased in the each month of posttreatment compared to pretreatment levels only in the experimental group (p<0.05). No abnormal finding was detected on digital rectal examination or in serum PSA, cholesterol, AST and ALT levels in either guoup. Sexual dysfunction was the most common problem prior to treatment, followed by metabolic, psychatric, musculoskeletal, neurologic, gastrointestinal and cardiopulmonary dysfunction, in order. After treatment, sexual dysfunction showed the most improvement in symptom score followed by cardiopulmonary, metabolic, cardiopulmonary, musculoskeletal, and gastrointestinal functions showed significantly more improved symptom scores in the experimental group than in the control group. The positive rate of Saint Louis University Questionnaire significantly decreased in the posttreatment compared to pretreatment levels only in the experimental group (p<0.05). The complications were nonspecific symptoms such as gastrointestinal problems and fatigue in 3 cases, and acne, urticatria and joint pain in 1 case of the experimental group. But, they were self-limited and did not cause drop-out. CONCLUSIONS: Androgen supplement therapy with oral testosterone undecanoate (Andriol(R)) enhanced the quality of life through the improvement of general body functions including physical functions and mental functions in men with testosterone deficiencies.