Mechanism of Xinfeng Capsules improving rheumatoid arthritis based on CD19~+B cells regulating FAK/CAPN/PI3K pathway / 中国中药杂志

To observe the effect of Xinfeng Capsules on rheumatoid arthritis (RA) B lymphocytes,inflammatory mediators,FAK/CAPN/PI3K pathway,in order to explore the mechanism of Xinfeng Capsules in improving clinical symptoms of RA.Joint and systemic symptoms of RA patients were observed,and laboratory indicators[hemoglobin (HGB),platelet count (PLT),erythrocyte sedimentation (ESR),immunoglobulin (Ig) G,Ig A,Ig M,rheumatoid factor (RF),anti-cyclic citrulline antibody (CCP-AB),C-reactive protein (CRP)]were detected.ELISA was used to detect serum interleukin (IL)-1β，IL-10,IL-33,chemokine 5 (CCL5),and vascular endothelial growth factor (VEGF).CD3~-CD19~+B cells were measured by flow cytometry.Western blot was used to detect FAK,p-FAK,CAPN,PI3K protein.The results showed that Xinfeng Capsules could significantly alleviate RA joint and systemic symptoms and improve clinical efficacy.And Xinfeng Capsules could increase HGB,decrease PLT,CCP-AB,CRP,ESR index,upregulate IL-10 expression,and down-regulate IL-1β，IL-33,CCL5,VEGF,CD3~-CD19~+B cells,FAK,p-FAK,CAPN,PI3K expressions (P<0.01).Based on the above results,Xinfeng Capsules may reduce the expression of CD3~-CD19~+,regulate the balance of inflammatory cytokines and chemokines,inhibit abnormal activation of FAK/CAPN/PI3K pathway,and improve clinical symptoms of RA.

Gene cloning, protein expression and examination of biological activity of chicken CD40L / 生物工程学报

To obtain chicken CD40L protein, the cDNA was prepared from chicken splenic cells and used as a template to clone and amplify CD40L by PCR. The target gene was cloned into pFastBac vector to construct a pFastBac-chCD40L donor plasmid. Recombinant plasmid was transformed into DH10Bac and recombinant Bacmid-chCD40L was obtained. The Bacmid-chCD40L plasmid was transfected into sf9 insect cells to obtain His-chCD40L protein. In addition, the target gene was cloned into pQM01 vector to construct a pQM01-chCD40L plasmid, recombinant plasmid was transfected into HEK 293T cells to obtain Strep-chCD40L protein. The chCD40L protein was purified by affinity chromatography, and the concentration of purified chCD40L protein was determined to be 0.01 mg/mL. Primary cells were isolated from the bursal tissue of 3-week old SPF chickens, and the chCD40L protein was added to the culture medium to stimulate cells. The chCD40L could bind to CD40 on B cells as examined by Western blotting, indirect immunofluorescence assay and flow cytometry, suggesting that chCD40L protein is biologically active. We successfully obtained chicken CD40L protein of biological activity, which laid the foundation in the in vitro culture of primary B lymphocytes for the isolation and diagnosis of virulent IBDV.

Efficacy of mandibular molar distalization by clear aligner treatment / 中南大学学报(医学版)

OBJECTIVES@#At present, the research on clear aligner of molar distalization mainly focuses on the upper jaw, while the research on mandibular molars is few.This study aims to evaluate the therapeutic effect of mandibular molars distalization with clear aligner via cone beam CT (CBCT) and Dolphin software.@*METHODS@#Twenty cases of mandibular molars with clear aligner were included according to the inclusion and exclusion criteria. CBCT was taken before treatment (T0) and when the first molar was moved in place (T1). Dolphin software was used to measure the effectiveness of molar distalization. Three-dimensional changes in direction and the impact on the incisors and facial soft and hard tissues were evaluated.@*RESULTS@#The effective rates of crown and root distalization of the second and first mandibular molars were 74%, 49%, and 71%, 47%, respectively. The second and first molars were both the distal buccal cusp with the largest distalization [(2.15 ± 0.91) mm and (1.85±1.09) mm], respectively, with significant difference between the T0 and T1 (@*CONCLUSIONS@#Clear aligner can effectively move mandibular molars farther, the crown is more effective than the root, and it is tilted. The second mandibular molar is more effective than the first mandibular molar in its distant displacement and three-dimensional changes. Molar distalization causes minor changes in mandibular incisors and facial soft and hard tissues.

Attenuation Mechanism of Aconti Lateralis Radix Praeparata Combined with Glycyrrhizae Radix et Rhizoma Based on Heart CYP450 Isozymes / 中国实验方剂学杂志

<b>Objective::To investigate the effect of Aconiti Lateralis Praeparata Radix combined with Glycyrrhizae Radix et Rhizoma on mRNA expression of cytochrome P450 (CYP450) enzymes in rat' s heart. <b>Method::Male Sprague Dawley rats were randomly divided into the control group, the inducer group (0.08 g·kg^-1·d^-1), the aconite group (0.5 g·kg^-1·d^-1), the licorice group (0.5 g·kg^-1·d^-1) and the compatibility group (aconite and licorice group 0.5 g·kg^-1·d^-1), with 6 rats in each group. In each group, the serum concentrations of aspartate transaminase (AST), creatine kinase (CK) andlactate dehydrogenase (LDH) were respectively measured. Hematoxylin-eosin(HE) staining was used to detect the pathologic changes in the heart tissue of the rats. The expression of several CYP genes were measured. <b>Result::After combination, the contents of AST, CK and LDH were lower than those of the aconite group. At the same time, the pathological results showed cardiac injury in rats in the aconite group, and licorice could alleviate the cardiac injury caused by aconite. At the level of gene transcription, the glycyrrhizae group could up-regulate the expressions of CYP2C11 and CYP2J3 to different degrees (<italic>P</italic><0.05), and the combination of licorice and aconite could up-regulate the expression of CYP2B1, CYP2C11 and CYP2J3 family (<italic>P</italic><0.05). It suggested that the combination could promote the formation of EETs in arachidonic acid(AA) metabolism, and the aconite could up-regulate the expressions of CYP4A1, CYP4A3, CYP4F5 and CYP4F6 in CYP4 family (<italic>P</italic><0.05), promote the formation of 20-hydroxyeicosatetraenoic acid(20-HETE). The combination of aconite and licorice can weaken the ability of aconite, down-regulate the expression of CYP4A3, CYP4F1, CYP4F5 and CYP4F6 family mRNA (<italic>P</italic><0.05), inhibit the formation of 20-HETE and reduce the cardiotoxicity caused by aconite. <b>Conclusion::The combination of licorice and aconite can regulate the expression of CYP450 enzyme in the heart, increase the production of EETs and the content of 20-HETE, and finally play a role in reducing toxicity.

Case-control study on temperament characteristics of children with autism spectrum disorders aged 3-7 years old in Harbin / 中国学校卫生

Objective@#To explore the differences of temperament characteristics between children with autism spectrum disorder (ASD) and normal children, and to provide evidence for early detection of ASD children and the development of personalized treatment plans.@*Methods@#In this case-control study, we enrolled 129 ASD children and 129 normal children aged 3-7 years. The Behavioral Style Questionnaire (BSQ) scale was used to assess the temperament.@*Results@#ASD children got higher scores in terms of "activity level", "withdrawal", "adaptability", "emotional nature", "persistence", and "response threshold" temperament dimensionality scores(P<0.05), and lower scores in terms of "rhythmical", "response intensity" temperament dimensionality scores than normal children(P<0.05). However, there was no significant difference in "attention dispersiveness" between ASD group and control group(P>0.05). Among the children in the ASD and control group, the proportion of each temperament type was "easy to raise temperament type" (41.8% vs 62.8%), "partially easy to raise temperament type" (31.8% vs 27.9%), "partially difficult to raise temperament type" (17.1% vs 6.2%), “slow-up-towarm temperament type" (7.7% vs 2.3%) and "difficult to raise temperament type" (1.6% vs 0.8%).Statistical analysis showed that the rate of "easy to raise temperament type" was lower than that in normal children(P<0.05), while the rates of "partially easy to raise temperament type", "partially difficult to raise temperament type", "difficult to raise temperament type", and "slow-up-to-warm temperament type" in ASD children were higher(P<0.05).@*Conclusion@#There was significant difference in temperament characteristics between ASD children and normal children. The evaluation of temperament type contributes to early detection of ASD children and provides a reference for their behavioral correction.

Effect of Pudilan Xiaoyan Oral Liquid on Acute Lung Injury Rat Induced by Lipopolysaccharide / 中国实验方剂学杂志

Objective:To explore the effect and mechanism of Pudilan Xiaoyan oral liquid(PDL) on the acute lung injury rat induced by lipopolysaccharide (LPS). Method:The 72 Wistar rats were randomly divided into control group, model group, dexamethasone group, PDL 7, 3.5, 1.75 g·kg-1·d-1 group according to body weight.The acute lung injury model was made through inhalation with lipopolysaccharide in the model group, hexadecadrol group, PDL 7, 3.5, 1.75 g·kg-1·d-1 group.To examining each rat alveolar lavage fluid (BALF) of the total number of white blood cells, enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of nuclear transcription factors-kappa B (NF-κB) and interleukin-10 (IL-10).Hematoxylin-eosin(HE) staining was used to observe morphological changes of lung tissue and explore different doses of PDL effect on acute lung injury in rats. Result:Compared with model group, the account of leukocyte in BALF decreased significantly in PDL 7 g·kg-1·d-1 group and PDL 3.5 g·kg-1·d-1 group (PκB significantly decreased in PDL 7, 3.5, 1.75 g·kg-1·d-1 group (P-1·d-1 group (P-1·d-1 group, the inflammation, edema and congestion in lung tissue reduced (PConclusion:PDL has a significant protective effect on the inflammation of acute lung injury model, and its mechanism is related to the expressions of NF-κB and IL-10. PDL could also repair the injury of lung in acute lung injury model.

Pharmacodynamic Evaluation of Tanreqing Inhalation Solution / 中国实验方剂学杂志

Objective: To clarify the antitussive, expectorant, antipyretic and anti-inflammatory effects of Tanreqing inhalation solution, and provide basis and data support for further research and development of this preparation. Method: The methods of cough induced by ammonia and tracheal phenol red excretion were used to observe the antitussive and expectorant effects of Tanreqing inhalation solution in mice. The fever model of rats was established by intraperitoneal injection of bacterial lipopolysaccharide(LPS) to observe the antipyretic effect of the Tanreqing inhalation solution, the acute pneumonia model of rats was established by atomizing LPS inhalation, and the anti-inflammatory effect of Tanreqing inhalation solution was observed. Result: Tanreqing inhalation solution could reduce the number of coughs in mice induced by ammonia water, increase the amount of phenol red excretion in mouse trachea, decrease the levels of body temperature and its related regulatory factors of prostaglandin E2(PGE2) and cyclic adenosine monophosphate(cAMP) of rats induced by LPS, decrease the white blood cell(WBC) count and the neutrophil ratio(NEUT) in bronchoalveolar lavage fluid(BALF) of rats with LPS-induced acute pneumonia, and reduce the levels of nuclear transcription factor-κB(NF-κB) and interleukin-1β(IL-1β) in lung tissue. Conclusion: Tanreqing inhalation solution has obvious antitussive, expectorant, antipyretic and anti-inflammatory effects, which is worthy of further development and promotion.

Effect of Different Administration Methods with Bleomycin on Pulmonary Fibrosis in Rats / 中国实验方剂学杂志

Objective:Compare the effects of 3 administration methods (tracheal perfusion, tail vein injection and aerosol inhalation) with bleomycin (BLM) in inducing pulmonary fibrosis in rats, in order to find out the optimal administration methods. Method:Eighty sprague-dawley (SD) male rats with SPF were randomly divided into aerosol inhalation blank group, single tracheal perfusion group(10 mg·kg-1), multiple tracheal perfusion group(5 mg·kg-1), single intravenous injection group(150 mg·kg-1), multiple intravenous injection group(50 mg·kg-1), single aerosol inhalation group (30 min)and multiple aerosol inhalation group(30 min). The mortality and body weight of rats in each group were observed at 7 d, 14 d and 28 d after the administration. And 28 days later after the administration, the lung coefficients of rats in each group were observed, paraffin sections were prepared, hematoxylin-eosin staining (HE) and Masson staining were performed, and the contents of hydroxyproline (HYP) and plasminogen activator inhibitor-1 (PAI-1) in lung tissues were detected by enzyme-linked immunosorbent assay (ELISA), so as to evaluate the alveoli inflammation and pulmonary fibrosis of rats in each group. Result:Compared with the aerosol inhalation blank group, the rats in the trachea perfusion group had the highest mortality among the drug treatment groups. The pulmonary coefficients of rats in the multiple intravenous injection group and the multiple inhalation group were significantly higher than those in the blank group(PPPConclusion:Bleomycin was inhaled repeatedly to establish pulmonary fibrosis model. The pathological injury and physiological indexes of the model rats were relatively stable, which conforms with the evolution process of pulmonary fibrosis.

Investigation of Physicochemical Properties and Determination Methods of Liposomal Entrapment Efficiency of Dichroa Alkali Hydrochloride / 中国实验方剂学杂志

Objective: To study on the physical and chemical properties of dichroa alkali hydrochloride and to establish a method for the determination of entrapment efficiency of dichroa alkali hydrochloride liposomes. Method: HPLC was used to determine the content of dichroa alkali hydrochloride with mobile phase of acetonitrile-water-triethylamine-glacial acetic acid(9:91:0.35:0.75) and detection wavelength at 265 nm.The oil-water partition coefficient of this compound in different pH range was measured by shake flask method.The stability of the dichroa alkali hydrochloride in phosphate buffer solution with different pH after sterilization at 125℃ for 30 min was investigated.Ammonium sulfate gradient method was used to prepare dichroa alkali hydrochloride liposomes.The microcolumn was prepared by dextran gel and cation exchange resin,respectively.Then the free drug and liposome were separated by centrifugation,the drug content was measured,and the encapsulation efficiency was calculated.The t-test was performed using SPSS 20.0 software,the differences between these two methods were compared. Result: In the pH 6-9,the oil-water partition coefficient of dichroa alkali hydrochloride increased with increasing of pH,which was between 0.016 and 1.44;the recovery rate of dichroa alkali hydrochloride after sterilization was 37.16%-57.91%.Between the dextran gel microcolumn centrifugation and the cation exchange resin microcolumn centrifugation,there was no significant difference in the entrapment efficiency of the liposomes. Conclusion: Dichroa alkali hydrochloride is suitable for preparation of liposomes.However,its stability is not ideal,so the experimental temperature should be strictly controlled in the preparation process.Dextran gel microcolumn centrifugation and cation exchange resin microcolumn centrifugation can be used to determine the entrapment efficiency of dichroa alkali hydrochloride liposomes,and the cation exchange resin microcolumn centrifugation is suggested after comparison.

Effects of human leucocyte antigen-G expression on invasion and proliferation of chorionic trophoblastic cell line JEG-3 / 中华妇产科杂志

Objective To investigate the effects of human leukocyte-associated antigen-G (HLA-G) expression in silencing trophoblast cell line JEG-3 under normal and hypoxic conditions on invasion and proliferation of JEG-3 cells. Methods Inhibition of HLA-G expression in JEG-3 cells by transfection of small interfering RNA (siRNA),the transfected JEG-3 cells were divided into 4 groups: normoxia control group, hypoxia control group, normoxia inhibition group and hypoxia inhibition group. The levels of HLA-G mRNA and protein in 4 groups of cells were detected by real-time quantitive PCR and western blot. The proliferation activity and invasion ability of 4 groups of cells were determined by methylthiazolyl tetrazolium (MTT) assay and invasion assay.Results (1) Real-time quantitive PCR technology showed: the level of HLA-G mRNA in the hypoxic inhibition group (0.220±0.050) was significantly different (P<0.05), when compared with that in the hypoxic control group (0.630±0.030) and normoxic inhibition group (0.400± 0.020). (2) Western blot analysis showed: the expression level of HLA-G protein in the hypoxic inhibition group was 0.260±0.010, statistically different from that in the hypoxic control group (0.850±0.100) and the normoxic inhibition group (0.560±0.020; P<0.05).(3) MTT showed: proliferative activity of JEG-3 cells in the normoxic inhibition group was 0.490 ± 0.070, the ability of cell proliferation was reduced. When compared with that in the normoxic control group (0.850±0.050), the differences was statistically significant (P<0.05). The proliferative activity of JEG-3 cells in the hypoxic inhibition group (0.330±0.070) was lower than that in the normoxic inhibition group (0.490±0.070), and there was a significant difference (P<0.05). (4) Invasion assay showed: compared with the normoxic control group (98±7), the invasive ability of JEG-3 cells in the normoxic inhibition group (73 ± 7) was weakened, and the difference was statistically significant (P<0.05). The number of transmembrane cells (52±11) of JEG-3 cells in the hypoxic inhibition group was lower than that in the hypoxic control group (72±7), and the difference was statistically significant (P<0.05). Compared with the normoxic inhibition group, the invasion ability of JEG-3 cells in the hypoxic inhibition group decreased, and the difference was statistically significant (P<0.05). Conclusion Under hypoxia, using siRNA technology to down-regulate the expression of HLA-G may affect the proliferation and invasion ability of trophoblast cells, which may be involved in the occurrence of hypertensive disorder of pregnancy.

Exploration of the Construction of Medical Alliance Under Public and Private Partnership / 中国医院管理

The medical alliance is an important carrier for the construction of hierarchical medical treatment system.In the study,the medical alliance under public and private partnership is explored and analyzed,the necessity of private medical institutions participating in the medical alliance construction is expounded.The feasibility of the construction of medical alliance under public and private partnership is illustrated from three aspects,such as policy,theory and practice.Based on the policy background and market environment of China,challenges of the construction of public and private partnership is analyzed,and the corresponding suggestions are proposed to provide ideas for the long-term development of medical alliance in China.

The effect of acupuncture combined with auricular application for puerperal hypogalactia / 中国基层医药

Objective To observe the effect of acupuncture and auricular application in the treatment of puerperal hypogalactia.Methods 300 patients with puerperal hypogalactia,according to the principle of random, balanced,together,were divided into 3 groups:Chinese herbal medicine group and acupuncture combined with auricular group and massage group,each group in 1 00cases.Collected patients'informed consent,and obtained the hospital ethics committee approval.The clinical curative effect was observed.Results After 3 days,in the Chinese herbal medicine group 34 cases recovered,24 cases were markedly effective,efficient in 1 5 cases,27 cases were invalid.In the massage group 7 cases recovered,1 3 cases had marked effect,1 5 cases were effective,ineffective in 65 cases.In the acupuncture combined with auricular group 73 cases cured,1 3 cases were markedly effective,effective in 9 cases, 5 cases were ineffective.The total effective rate between the acupuncture combined with auricular group and Chinese herbal medicine group had significant difference(χ2 =9.600,P <0.01 ).The total effective rate between the acupuncture combined with auricular group and the massage group had significant difference (χ2 =1 3.400,P <0.01 ).The total effective rate between Chinese herbal medicine group and massage group had significant difference (χ2 =6.400,P <0.01 ).Conclusion Acupuncture and auricular application has obvious curative effect for patients with postpartum hypogalactia.

Effects of IL-1β on expression of AQP4 and its role in attack of seizure / 实用医学杂志

Objective To observe the effects of interleukin-1β (IL-1β) and pentylenetetrazol (PTZ) induced acute epilepsy and the dynamic expression of aquaporin-4 (AQP4) in hippocampus. To explore the role of IL-1β in the pathogenesis of epilepsy by regulating AQP4. Methods All rats were randomly divided into control group, IL-1β group, PTZ group, IL-1ra + PTZ group and dexamethasone + PTZ group. Observe the behavior of the rats within 60 minutes after injection and record seizure score in each group. Then immunohistochemistry and RT-qPCR were used to detect the expression of AQP4 at at 6 , 12, 24 and 36 h. Results Almost of rats in IL-1β group and PTZ group showed severe degree seizure. The rats in control group and dexamethasone + PTZ group showed no obvious seizure. The seizure of rats were more remarkable serious in PTZ group than that in the IL-1ra + pentylenetetrazole group (P < 0.05). Immunohistochemistry and RT-qPCR Show: the expression of AQP4 in hippocampus in PTZ group increased gradually after 12 h (P < 0.05), then reached in the peak after 24 h (P < 0.001). The expression of AQP4 in IL-1ra + PTZ group was lower compare with PTZ group in each time (P < 0.05). Although the expression of AQP4 in dexamethasone + PTZ group higher than the control group, it was not significantly different (P < 0.05). Conclusion The proinflammatory cytokine IL-1β break the balance of water in brain and increasing the concentration of extracellular excitatory amino acids or ions by upregulate the expression of AQP4 in order to promote the excitatory of neurons.

On-demand versus on-time dosing of tadalafil for erectile dysfunction: a prospective multi-center study / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the safety and efficacy of tadalafil on demand and on time in men with erectile dysfunction.</p><p><b>METHODS</b>We conducted a multi-centered randomized controlled study on 120 ED males, who were assigned to take tadalafil at 10 mg/ 20 mg on demand before sexual activity and at the same dose on time twice a week for 8 weeks. Before and at 4 and 8 weeks after treatment, and 1 month after withdrawal, we obtained the scores on IIEF-5, ED Inventory of Treatment Satisfaction (EDITS) and the short form of Psychological and Interpersonal Relationship Scales (SF-PAIRS) , and compared the safety and efficacy of medication between the two groups of patients.</p><p><b>RESULTS</b>Totally, 110 patients accomplished the trial, 56 in the on-time and 54 in the on-demand group. At 4 and 8 weeks of medication and 1 month after withdrawal, the IIEF-5 scores were improved in both the on-time and on-demand groups, even more significantly in the former than in the latter at 8 weeks of treatment (21.6 +/- 2.9 vs 18.5 +/- 1.7) and 1 month after withdrawal (20.9 +/- 2.1 vs 17.9 +/- 2.3) (P < 0.05). The EDITS scores were significantly higher in the on-time than in the on-demand group at 8 weeks of treatment (31.7 +/- 6.9 vs 28.6 +/- 5.8) and 1 month after withdrawal (30.6 +/- 4.7 vs 27.9 +/- 6.5) (P < 0.05). The scores on the sexual self-confidence, spontaneity and time-concern domains of SF-PAIRS were remarkably improved after medication as compared with the baseline (P < 0.05), even more significantly in the on-time than in the on-demand group at 1 month after withdrawal. Both dosing schedules were well tolerated and no significant differences were observed in safety between the two groups.</p><p><b>CONCLUSION</b>On-time dosing of tadalafil is efficacious and well tolerated in the treatment of ED, and has an even better effect than on-demand dosing at 8 weeks of medication and 1 month after withdrawal.</p>

Vacuum therapy for erectile dysfunction that fails to respond to PDE-5i: report of 70 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>To assess the efficacy and safety of the Osbon ErecAid Vacuum Therapy System in the treatment of erectile dysfunction (ED) that fails to respond to PDE-5i.</p><p><b>METHODS</b>This study included 70 cases of ED not responding to the medication of PDE-5i. The patients were aged 25 -66 (38.9 +/- 9.1) years. Under our direction, the patients used the Osbon ErecAid Vacuum Therapy System for a month to increase blood oxygenation to the corpora cavernosa, and then employed the vacuum constriction device (VCD) to maintain penile erection for sexual intercourse. We compared their IIEF-5 scores and their and their partners' sexual satisfaction before and 3 months after treatment, and evaluated the safety and effect of the vacuum therapy.</p><p><b>RESULTS</b>The mean IIEF-5 scores of the patients were significantly increased from 10.2 +/- 4.0 before treatment to 13.3 +/- 4.7 at 3 months after the vacuum therapy (P < 0.05). The rate of therapeutic effectiveness was 77.1% (54/70), and the rates of sexual satisfaction of the patients and their partners were 64.3% (45/70) and 65.7% (46/70),</p><p><b>CONCLUSION</b>Vacuum therapy for ED that does not respond to PDE-5i can significantly improve the patients' erectile function as well as their and their partners'sexual satisfaction, and therefore deserves clinical application.</p>

Three-step sperm retrieval for 73 non-obstructive azoospermia patients / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the value and clinical application of the Three-Step Sperm Retrieval method in improving the sperm retrieval rate for non-obstructive azoospermia (NOA) patients.</p><p><b>METHODS</b>Seventy-three NOA patients underwent Three-Step Sperm Retrieval in the following order of procedures: testicular fine needle aspiration (TFNA), testicular sperm extraction (TSE), and microdissection testicular sperm extraction (MD-TSE). The testicular tissue obtained from each step was observed for spermatozoa under the 400-fold inverted microscope. If spermatozoa were found in one step, the operation would be stopped; otherwise, the next step would be carried out. The testicular tissue was subjected to pathological examination.</p><p><b>RESULTS</b>Spermatozoa was found in the testicular tissue in 38.4% of the cases (28/73) at TFNA as the first step, in 52.1% (38/73) at TFNA and TSE, and in 64.4% (47/73) at TFNA, TSE and MD-TSE. Pathological examination showed 25 of the cases to be Sertoli cell-only syndrome, 21 to be sperm maturation arrest and the other 27 to be hypospermatogenesis, in which spermatozoa were found in 10, 14 and 23 cases, respectively.</p><p><b>CONCLUSION</b>The Three-Step Sperm Retrieval method can significantly improve the sperm retrieval rate for NOA patients. And the sperm retrieval rate is associated with the pathological type of the testicular tissue, a higher rate with hypospermatogenesis.</p>

Appliance of microsurgery in the treatment of male infertility / 中华泌尿外科杂志

Objective To discuss the application of microsurgery in the treatment of male infertility.Methods From March 2007 to March 2012,there were totally 853 infertile men received microsurgical treatments in our department.Among them,344 patients with unilateral or bilateral varicocele underwent microsurgical varicocelectomy,60 underwent vasovasostomy (VV) and 192 underwent vastoepidystomy (VE)in microsurgical methods due to obstructive azoospermia.257 non-obstructive azoospermia (NOA) patients were performed microdissection of testicular sperm extraction (MD-TESE),at the same time,pathologic examination was done.Results ①For the varicocele patients,the pre-operative sperm density was (10 ±6) × 106/ml,the progressive sperm percentage was (16 ± 9)％.The post-operative density was (15 ± 8) ×106/ml,the progressive sperm percentage was (28 ± 14)％.The natural pregnant rate was 10.8％ (37/344).②In 60 patients undergone VV,the patent rate was 80.0％ (48/60),the natural pregnant rate was 35.0％ (21/60).In 192 VE patients,the patent rate was 53.1％ (102/192),the natural pregnant rate was 19.8％ (38/192).③In 257 NOA patients,the testicular volume,sperm retrieval rate of MD-TESE was significantly higher than that of conventional testicular sperm extraction (60.3％ vs.38.1％).Conclusion The microsurgery techniques in male infertility treatments could have some advantages such as explicit effects and decreased injuries.

Reconstructed seminiferous tubules from ectopic grafting of testicular cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the ectopic grafts of mouse testicular cells by observing the reconstruction of seminiferous tubules, colonization of spermatogenic cells and spermatogenesis using immunodeficient mice as recipients.</p><p><b>METHODS</b>The testes of newborn male ICR mice were digested to obtain single cell suspension. The cells were then mixed with matrigel and subcutaneously grafted into the dorsal region of the male nude mice. The mice were castrated after the operation and the grafts were dissected from 5 of the nude mice at 4, 6, 8 and 10 weeks, respectively. The success rates of transplantation and the graft diameters were calculated, and the structure of the reconstituted seminiferous tubules, colonization of the germ cells and spermatogenesis were observed by HE staining and immunohistochemistry.</p><p><b>RESULTS</b>All the mice recipients survived after the testicular cell transplantation. Within 10 weeks after the operation, tissue masses could be observed, with the diameter increased from (3.91 +/- 0.71) mm at 4 weeks to (6.69 +/- 0.50) mm. Neovascularization was detected at the surface of the masses and seminiferous tubule structures found in the grafts. The germ cells that developed from spermatogonia to round spermatids were observed, but with no sperm in the tubules. Germ cells, Sertoli cells and Leydig cells were identified by immunochemical detection of Mvh, Gata4 and P450Scc in the grafts at 8 weeks.</p><p><b>CONCLUSION</b>Seminiferous tubules could be ectopically reconstructed from suspension of neonatal mouse testicular cells. Ectopic grafting provided a preferable model for the studies on testis tissue engineering and interactions between testicular cells during testicular development and spermatogenesis.</p>

Association of SNP276 in adiponectin gene with type 2 diabetes mellitus and insulin sensitivity / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the distribution of SNP276 in adiponectin gene in Chinese Hans and its impact on type 2 diabetes mellitus and insulin sensitivity.</p><p><b>METHODS</b>The study population consisted of 417 Chinese Hans residents in Anhui province, including 141 subjects with normal glucose tolerance (NGT) and 276 with type 2 diabetes (T2DM). The islet beta-cell insulin secretion and tissue insulin sensitivity were assessed by formulae of homeostasis model assessment (HOMA-IR & HOMA beta). Firstly, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to determine whether variation exists in APM1. Then, exact variation was detected by automated DNA direct sequencing.</p><p><b>RESULTS</b>The genotypes of APM1 SNP276 were 0.489 GG, 0.418 GT and 0.092 TT and the major allele was G (frequency=0.699) in subjects with NGT. The distributions of genotypes and alleles of SNP276 both displayed significant difference between NGT and T2DM groups (P=0.031 and 0.013). The SNP276 non-TT (TG+GG) genotype was associated with increased risk of T2DM (OR=2.447, 95%CI: 1.067-5.612, P=0.035). In T2DM group, the subjects with SNP276 GG or GT genotype had higher body mass index, body fat content, fasting plasma glucose and HOMA-IR than did those with TT genotype (P < 0.05 or P < 0.01). Besides, GG genotype had higher systolic blood pressure (P=0.021). In NGT group, SNP276 non-TT carrier had increased body mass index, body fat content, waist hip ratio, fasting plasma insulin, oral glucose tolerance test 2 h plasma insulin and HOMA-IR when compared with TT genotype (P < 0.05 or 0.01).</p><p><b>CONCLUSION</b>SNP276 in APM1 was associated with T2DM and insulin sensitivity.</p>