Objective To explore the association of anti-mullerian hormone(AMH)in seminal plasma and serum with sperm counts and energy for male.Methods For 215 cases of healthy male selected from our reproductive clinic,with women′s reason for infertility,seminal plasma and serum AMH were detected,as semen parameters(sperm density,living rate,vitality and malformation rate),6 items of serum sex hormone.In seminal plasma and serum AMH respectively as the dependent variable,using multiple line-ar regression model to explore its quantitative relation with semen parameters and sex hormone levels.Results 215 cases were en-rolled,aged 34.28±5.70 years,while the median of the seminal plasma AMH was 0.47,quartile 0.05-3.09 pmol/ejaculation.The median of the serum AMH was 53.07,quartile 32.32 -72.20 pmol/L.Through multiple linear regression analysis,after adjusted by age and BMI,the seminal plasma of AMH and total number of sperm,sperm concentration,dynamic motility,total sperm activi-ty,serum inhibin B were positively correlated(P <0.05);The correlation between sperm morphology and other serum sex hormone had no statistical significance(P >0.05);Serum AMH negatively correlated with serum FSH,with serum inhibin B positively(P <0.05);Seminal plasma in various parameters and other related serum sex hormone had no statistical significance(P >0.05).Conclu-sion The seminal plasma of AMH were positively correlated with sperm concentration,sperm counts,sperm vitality,with the asso-ciation for serum AMH not yet found.

Objective To explore a new surgical procedure for phallic reconstruction. Methods Since Dec 2003 to Nov 2006,4 patients with penile loss were reconstructed by transferring a deep in-ferior epigastric perforator(DIEP)flap with implanting the 12th costal cartilage. Results All of the flaDs survived successfully.With the follow-up from 5 months to 3 years,the appearance oi the neope-nis and donor-site scar was satisfactory. No one developed fistula or urethral stenosls, All patients were reported the return of erogenous sensibility of the neopenis leading to orgasm and achmving Pene-tration during sexual intercourse. Conclusions This new procedure may provide an esthetlcaIlY ac-ceDtable and functional neophallus.It might be a new alternative in penile reconstruction.

BACKGROUND: Classic isolation method of bone marrow mesenchymal stem cells (BMSCs) is Percoll density gradient centrifugation. Blood cell component was removed. However, this method is complicated. Preparation density was needed when isolating dog bone marrow. Moreover, centrifugation was frequent, which had a great damage to cells. OBJECTIVE: To establish methods of the isolation, proliferation, culture and osteoinduction of canine BMSCs, and observe the in vitro proliferation and ability to osteoinduction differentiation. METHODS: 10 mL bone marrow was extracted from dog posterior superior iliac spine, heparin anticoagulation, diluted using Hanks juice, treated with 1.077 g/mL Ficoll solution 3 mL, and centrifuged at 2 000 r/min for 20 minutes. Karyocytes were absorbed to form white cloudlike layering interface, and then centrifuged twice using DMEM supplemented with fetal bovine serum, incubated at 12×10~4/cm~2 at 37 ℃ in a 5% CO_2 incubator. Following subculture, cells were incubated in DMEM containing dexamethasone, β-sodium phosphoglycerol and ascorbic acid 2-phosphate. Immunocytochemical staining and immunofluorescence staining were utilized to detect osteocalcin, osteopontin and type Ⅰ collagen expression in osteoblasts. Alkaline phosphatase staining and alizarin red staining were performed. RESULTS AND CONCLUSION: 1.077 g/mL Ficoll density gradient centrifugation was used to isolate karyocytes that were significant compared with Percoll solution. Obtained BMSCs had high purity, good growth and the mean doubling time was 24 hours. Following in vitro osteogenic incubation of dog BMSCs, osteocalcin, osteopontin and type Ⅰ collagen showed positive expression. Alkaline phosphatase staining demonstrated bluish-green cytoplasm. Alizarin red staining showed red nodes in extracellular matrix, and could differentiate into osteoblasts in vitro.

BACKGROUND: There are currently many cryopreservation methods for the aminotic membrane, which have varying effects on the ultrastructure and biological activity of amniotic membrane, but on no one is effective.
OBJECTIVE: To compare the effects of different cryopreservation methods on the ultrastructure and viability of aminotic membrane and to seek the ideal cryopreservation method.
METHODS: Aminotic membrane separated from the fresh placenta was preserved respectively with deep-frozen cryopreservation and vitrification, and everyway was run for 3 and 6 months. Fresh aminotic membrane was used as control. The ultrastructure of aminotic membrane was observed by transmission electron microscopy, and the viability of aminotic membrane was assessed by microcomputer analysis system for biological oxygen consumption, and immunohistochemical staining combined with image analysis system was used for lactate dehydrogenase activity.
RESULTS AND CONCLUSION:After 3 and 6 months of crypreservation, the damage to the ultrastructure of aminotic membrane by vitreous cryopreservation was slighter than that of amniotic membrane cryopreserved at-80℃. Compared with the fresh aminotic membrane, the gray value of lactate dehydrogenase and partial pressure of oxygen were significantly decreased in the cryopreserved aminotic membrane by deep-frozen cryopreservation at 3 and 6 months (P < 0.05) and by vitreous cryopreservation at 6 months (P < 0.05), but there was no statisticaly significant difference in the change rate of oxygen partial pressure and the gray value of lactate dehydrogenase between the fresh aminotic membrane and the cryopreserved aminotic membrane by vitreous cryopreservation at 3 months. The present study led to the conclusion that vitreous cryopreservation protocol alows to not only maintain the integrity of AM, but also to preserve the viability of the cels. So the vitreous cryopreservation is superior to the deep-frozen cryopreservation for cryopreservation of aminotic membrane.

Objective To investigate the expression and clinical significance of DJ-1 protein in the endometrial carcinoma and delineate its functional roles in endometrial carcinoma progression.Methods Expression of DJ-1 was examined with immunohistochemical staining of 69 paraffin-embedded endometrial carcinoma samples who underwent surgery at Hunan province Tumor Hospital.Kaplan-Meier analysis was performed to describe the survival curve.Univariate and multivariate analyses were performed with COX proportional hazard model.Results (1) The expression of DJ-1 protein in endometrial carcinoma was significantly higher than that in normal endometrial tissues (P ＜ 0.05).(2) There was positive correlation between the high expression of DJ-1 protein in endometrial carcinoma samples and the pathological grade,the International Federation of Gynecology and Obstetrics (FIGO) stage,depth of myometrial invasion,or lymph node metastasis (P ＜ 0.05).(3) Kaplan-Meier analysis indicated that the cumulative 5-year survival rate in the low-DJ-1 expression group was 80.8％,whereas it was only 51.2％ in the high-DJ-1 expression group (P ＜ O.05).COX analysis indicated that the expression of DJ-1 was not the endometrial carcinoma independent prognostic factors for overall survival time (P ＞ 0.05).Conclusions The high expression of DJ-1 protein in endometrial carcinoma samples demonstrated a significantly positive correlation with the pathological grade,FIGO stage,depth of myometrial invasion,and lymph node metastasis.However,DJ-1 is not enough to be independent prognostic factor for overall survival time.

Objective:To determine mechanisms for the role of high mobility group box-1 (HMGB1) to platinum based chemo-sensitivity in cervical cancer.Methods:Using Western blot,we examined the effect of cisplatin on expressions of LC3,Beclin1 and P62 in cervical cancer.After treated with autophagy inhibitor and/or cisplatin in HeLa and CaSki cells,cell counting kit-8 (CCK-8) assay was used to evaluate the chemo-sensitivity.The expression of LC3,Beclin1 and P62 were detected by Western blot.We established HeLashHMGB1,CaSki-shHMGB1 and HeLa-CTR,CaSki-CTR stable cell lines.CCK-8 assay was used to determine the chemosensitivity in these cell lines.Using Western blot,we examined the correlation among the expressions of HMGB1,LC3,Beclin1,and P62.Results:With the increase of cisplatin,the expression of LC3 and Beclin1 was increased,while the expression of P62 was decreased in HeLa and CaSki cell lines.Combination of cisplatin with autophagy inhibitor could increase the cellular sensitivity to cisplatin (P＜0.05).The expression of HMGB1 was related to chemo-sensitivity and autophagy related protein in cervical cancer cells.HMGB1 was positively correlated with LC3 and Beclin1 while negatively correlated with P62 (P＜0.05).Conclusion:HMGB 1 might play an important role in chemo-sensitivity via regulating autophagic activity.The combination of cisplatin with autophagy inhibitor might become a possible strategy for cervical cancer.HMGB1 might serve as a potential biomarker for predicting chemo-therapeutic responses.

Objective To investigate the difference in intestinal flora among patients with esophageal squamous cell carcinoma and normal population and to provide a basis for the early diagnosis of esophageal squamous cell carcinoma as a marker. Methods DNA was extracted from biopsy tissue samples of 30 patients with esophageal squamous cell carcinoma (observation group) and 25 healthy people (control group) by microbial amplification sequencing. The integrity and quality of DNA were detected. The composition and abundance of intestinal flora in the samples of the two groups were determined. Results A great similarity in beta diversity was found between the two groups, but some differences were also observed. The relative abundance of Proteobacteria and Verrucomicrobia in the observation group was higher than that in the control group (P<0.05). The relative abundance of Megamonas in the observation group was lower than that in the control group (P=0.025). Conclusion Strengthening the study on the changes in intestinal flora among patients with esophageal squamous cell carcinoma may be of great significance for its prevention and treatment.

Objective:To clarify the incidence and the related risk factors of postoperative delirium in liver transplantation (LT) recipients to provide rationales for early identification of delirium and constructing the related models.Methods:The authors used the "肝移植""移植术""肝移植手术""肝脏移植""移植肝""谵妄""谵语""危险因素""相关因素""影响因素"and "liver transplantation""liver transplant""delirium""delirious""delirium confusion""risk factors""relevant factors""root cause analysis"as the Chinese and English keywords, searching Wanfang data, China Biomedical Literature Database, CNKI, PubMed, Embase, Web of Science, Cochrane Library, BMJ and the literature for the incidence or risk factors of postoperative delirium in LT recipients. The researchers independently performed literature screening, methodological evaluation and data extraction. And RevMan 5.4 and State16.0 software were employed for data processing.Results:A total of 19 articles involving 5003 samples were retrieved and 22 risk factors identifies. Meta-analysis showed that the incidence of POD was 23%(1151/5003). The statistically significant risk factors included preoperative blood ammonia concentration >46 mmol/L ( OR=3.51, 95% CI: 1.53-8.09, P<0.001), model for end-stage liver disease (MELD) score >15 points ( OR=4.24, 95% CI: 2.51-7.16, P<0.001), preoperative hepatic encephalopathy ( OR=3.00, 95% CI: 2.09-4.31, P<0.001), preoperative dosing of diuretics ( OR=2.36, 95% CI: 1.38-4.04, P<0.001), history of alcoholism ( OR=3.16, 95% CI: 1.06-9.40, P=0.040), longer anhepatic period ( OR=1.04, 95% CI: 1.03-1.06, P<0.001) and elevated aspartate transaminase concentration at Day 1 post-operation ( OR=1.33, 95% CI: 1.15-1.53, P<0.001). Conclusions:Preoperative blood ammonia concentration >46 mmol/L, MELD score >15, hepatic encephalopathy, dosing of diuretic, a history of alcoholism, longer anhepatic period and elevated aspartate transaminase at Day 1 post-operation are risk factors for postoperative delirium after LT. Postoperative reintubation is not a risk factor for postoperative delirium.

Objective To explore the safety and efficacy of intrathecal administration of adipose stem cells de-rived from bioactive secretome (ASCBS)in treatment of whiter matter injury (WMI)in the preterm infants. Methods Sixty - three cases of WMI were recruited according to the uniform standards from multiple medical centers and they were divided into 3 gestational age (GA)subgroups,which were 21 cases in group A (GA 24 - 28 + 6 ),20 cases in group B (GA 29 - 32 + 6 ),and 22 cases in group C (GA 33 - 36 + 6 ). The patients were randomly divided into treatment groups and control groups by tossing coins. The treatment groups received lumbar puncture followed with ASCBS intra-thecal injection once daily for 3 consecutive days. Follow - up study included Neonatal Behavioral Neurological Assess-ment (NBNA)at term - equivalent age and neurodevelopment at corrected age of 6 - month. Neurodevelopment was assessed by using the Bayley Scales of Infant Development and Peabody Developmental Motor Scale. The survival rates, NBNA scores,mental development index (MDI),psychomotor develop index (PDI),total motor development quotient, gross motor development quotient and fine motor development among each subgroup were compared. Results Sixty -three cases were recruited,including 31 in the treatment group and 32 in the control group. Only 1 case in the treatment groups lost in the follow - up. No clinical side effects were found in the treatment groups. There was no significant diffe-rence in the survival rate and complication in the preterms in all subgroups of the treatment group and control group (all P > 0. 05). The gross and total motor development quotient in the treatment group A was higher than that in the control group A(gross motor development quotient:98. 330 ± 6. 282 in treatment group A,90. 330 ± 3. 777 in control group A, P = 0. 040;total motor development quotient:97. 330 ± 4. 803 in treatment group A,91. 000 ± 4. 472 in control group A,P = 0. 023). The rest findings showed no significant difference between groups. Conclusion The treatment of WMI in preterm infants with ASCBS is safe and can promote the motor development of preterm infants with GA in 24 - 28 weeks.

Objective To find a method to repair nasal deformity of Tessier 3 cleft.Methods Five Tessier 3 cleft patients were treated from June 2010 to June 2015 in our department.Nasolabial groove flap or nasolabial groove flap accompanied by offside ala nasi groove flap were performed to correct the nasal deformity according to the defect of the ala.Results Two cases were treated with simple nasolabial groove glap,and 3 cases were treated with nasolabial groove flap accompanied by offside ala nasi groove flap.All the flaps survived with incisions of donor sites healed primarily.The color,texture and thickness of the flaps were similar to those of recipient sites after 1 year follow up.The scar in donor site was concealed and not obvious.The results showed good healing and good appearance,and the symmetry between the nostrils also seemed to be acceptable.Conclusions Nasolabial groove flap and nasolabial groove flap accompanied by offside ala nasi groove flap are good methods to correct nasal deformity of Tessier 3 cleft.The offside ala can be reduced at the same time and the symmetry between the nostrils can be achieved more easily.The operation is easily performed,the scar is acceptable and the results are good.