AIM: To study the effect of double gene transduction mediated by lentiviral vectors on the characteristics of human embryonic stem cells (hESCs). METHODS: Using the backbone from inducibal dual and excisable transgene vector (iDuet101) , lentiviral vectors overexpressing cytotoxic T lymphocyte - associated molecule - 4 immuno-globulin (iDuet101 - CTLA4Ig) , indoleamine 2, 3 dioxygenase (iDuet101 - IDO) , and ubiquitin - C promoter - lueifer-ase - ires - puromycin ( ULIP) were constructed and packaged according to the standard recombinant techniques. Human embryonic stem cells (hESCs) were first transduced with iDuet101, iDuet101 - CTLA4Ig or iDuet101 - IDO, then, after the selection, were transduced again with ULIP. The expression and function of the exogenous genes were detected. Immu-nohistochemistry, RT - PCR and flow cytometry were applied for detection of embryoid bodies ( EB) formation in vitro and in vivo teratoma formation. RESULTS: Double - transduced hESCs showed typical shape of cell clones and positive staining of tumor rejection antigen -1 - 60 ( Tra -1 - 60 ) and octomer transcription factor - 4 ( OCT - 4 ). The formation of EB was observed, in which a - fetoprotein (AFP), paired box gene 6 ( Pax6) and Musashi 1 ( MSI1) were positively expressed. The cells formed teratomas, and the luciferase signals existed until 28 days after xeno - transplantation. CONCLUSION : Double transduction of non - transcriptional factors mediated by lentiviral vectors does not affect the cell growth rate and their differentiation ability.

Objective To analyze the characteristics of erectile dysfunction(ED)in aged men.Methods Erectile dysfunction(ED)was diagnosed according to the International Index of Erectile Function(IIEF).78 aged men(average 65.9 yrs)as the study group,and 82 young patients(average 36.0 yrs)as the control group,all with ED,Were compared in complicating diseases,self-rating SDS scores,penile-brachial indexes(PBI),the time of achieving erection,and ejaculatory latency time.Results The main complicating diseases in study group were cardiovascular diseases(42 cases,53.9%),lower urinary tract symptoms(LUTS)(31 cases,39.7%)and diabetes(26 cases,33.3%).In the control group,the main complicating diseases were chronic prostatitis(52 cages,63.4%),premature ejaculation(PE)(32 cases,39.0%)and depression(12 csses,14.6%).SDS scores of study group and control group were(29.13±5.63)and(39.59±13.31),PBI were(0.78±0.12)and(0.91±0.06),the time of achieving erection were(13.85±5.75)min and(3.61±4.29)min,ejaculatory latency time were(7.03±5.35)min and(3.81±5.53)min.All with significant difference(P＜0.01).Conclusion Most of the ED old men were complicated with organic diseases,such as the time of achieving erection,PBI low scores and longer ejaculatory latency time.

[Objective]To compare the clinical outcomes of fresh embryo transfer of the in vitro fertilization/intracytoplasmic sperm injection and embryo transfer(IVF/ICSI-ET)in different age groups as well as in different responders using gonadotropin-re-leasing hormone agonist(GnRH-a)long protocol or GnRH antagonist(GnRH-ant)protocol.[Methods]A retrospective analysis was performed on 737 IVF/ICSI cycles,including 386 cycles of GnRH-a long protocol(group A)and 351 cycles of GnRH-ant protocol (group B),from August 28,2015 to December 31,2016. Then all the cycles were divided into sub-groups by ages and retrieved oo-cyte numbers:group a1(<38 years),group a2(≥38 years);group b1(n≤5),group b2(6≤n≤15),group b3(n>15). The basic information of patients and clinical outcomes were compared.[Results](1)Comparable results were obtained from group A and group B in these following variables such as fertilization rate,normal fertilization rate,biochemical pregnancy rate and miscarriage rage. But the stimulation period,the total gonadotropin(Gn)dosage,estradiol(E2)level and endometrial thickness on the day of human chorionic gonadotropin(hCG)administration,number of oocytes retrieved and mature oocytes,ovarian hyperstimulation syn-drome(OHSS)rate,implantation rate and clinical pregnancy rate were significantly higher in group A than group B(P<0.05),and significantly higher cancellation rate of fresh embryo transfer was observed in group B(P<0.001).(2)When divided by ages,no mat-ter in sub-group a1 or sub-group a2,the implantation rate was slightly lower in GnRH-ant protocol than in GnRH-a long protocol, although they failed to reach significant difference(sub-group a1:32.6%vs 39.8%,P=0.067;sub-group a2:9.7%vs 17.9%,P=0.066). The clinical pregnancy rate was comparable using these two protocols in sub-group a1(54.8%vs 50.4%,P=0.429),but it was significantly lower by using GnRH-ant protocol than GnRH-a long protocol in sub-group a2(19.6%vs 39.1%,P=0.021).(3) When divided by numbers of oocytes retrieved,the implantation rate was significantly lower when using GnRH-ant protocol in sub-group b1(13.1%vs 26.0%,P=0.026),but we failed to observe significant differences in other two sub-groups. The clinical preg-nancy rates were comparable in all sub-groups ,whereas differed considerably in sub-group b1 (36.6% vs 19.3%,P = 0.056).[Conclusion]Overall,the implantation rate and clinical pregnancy rate were higher in GnRH-a long protocol than those in GnRH-ant protocol. Nevertheless,GnRH-ant protocol could reduce the dosage of Gn,shorten the treatment duration,and effectively reduce the occurrence of OHSS. There were similar pregnancy outcomes in two protocols for normal responders and high responders ,while for advanced patients or other poor responders,the implantation rate and clinical pregnancy rate were higher in GnRH-a protocol.

AIM:To generate thalassemia-specific integration-free induced pluripotent stem cells ( iPSC) and to detect their ability of differentiation into hematopoietic precursors .METHODS:The plasmids pEB-C5 and pEB-Tg were transfected into the fibroblast cells from hemoglobin Bart ’ s hydrops fetalis ’ s skin by the method of nuclear transfection to reprogramm the cells into iPSC .The ability of the iPSC to differentiate into 3-germ layer cells was determined .The iPSC were cocultured with mouse OP 9 cells to differentiate into hematopoietic precursors and the hematopoietic precursor specific antigens were detected .RESULTS:The integration-free iPSC from hemoglobin Bart ’ s hydrops fetalis ’ s skin fibroblasts were successfully derived, and had the ability to differentiate into 3 germ layers.When cocultured with OP9 cells for 9 d, the positive rate of hematopoietic progenitor cell marker CD 34 was 18.7%, and the CD34 and CD45 double positive rate was 12.2%.CONCLUSION:Hemoglobin Bart ’ s hydrops fetalis ’ s skin fibroblasts can be successfully induced into “in-tegration-free” iPSC.This cell line has the ability to differentiate into 3 germ layers , and can be differentiated into hemato-poietic precursors when cocultured with OP 9 cells.

Objective To explore the clinical features and risk factors of poor prognosis in neonatal necrotizing enterocolitis(NEC).Methods A retrospective study was carried out in the infants with NEC admitted to 6 cooperative hospitals in Guangdong Province between January 2005 and December 2014.The clinical features and risk factors of poor prognosis in preterm and full-term infants diagnosed NEC,early onset and late onset NEC were analyzed.Results A total of 449 cases who met the criteria were admitted during the study time.The mortality was 23.6％ (106/449 cases),of which the preterm group was 24.6％ (58/238 cases) while the full-term group was 22.7％ (48/211 cases),the early onset group was 22.1％ (45/204 cases) while the late onset group was 24.3％ (57/235 cases).The median number of NEC onset in preterm group was 11 d after birth while the number of the full-term group was 6 d.Full-term infants who diagnosed NEC were more likely to manifest themselves as abdominal distension (52.1％ vs.42.0％,x2 =4.597,P =0.032),vomiting(36.5％ vs.17.2％,x2 =21.428,P =0.000) and bloody stool(30.3％ vs.21.4％,x2 =4.653,P =0.031);but in the onset of NEC,preterm infants more likely to have feeding intolerance (21.0％ vs.12.8％,x2=5.309,P =0.021).The early onset group of full-term NEC was much common in twins or multiplets(9.4％ vs.1.1％,x2 =6.226,P =0.013),which rate of surgical therapy was much higher (41.0％ vs.27.0％,P =0.036) and the breast-feeding rate before NEC was lower than the late onset group(14.5％ vs.32.6％,x2 =9.500,P =0.002),the differences were statistically significant.The gestational age and birth weight were bigger in the early onset group of preterm NEC[(33.8 ±2.5) weeks vs.(32.2 ±2.8) weeks,t =4.261,P =0.000;(2.1 ±0.5) kg vs.(1.7 ± 0.5) kg,t =4.735,P =0.000)],but length of stay was shorter than the late onset group (18.0 d vs.26.5 d,P =0.000).Logistic regression analysis showed that the risk factors of poor prognosis of full-term NEC were shock,peritonitis and sepsis;while risk factors of poor prognosis of preterm NEC were small for gestational age infant,pulmonary hemorrhage,shock,intestinal perforation and sepsis;the risk factors of poor prognosis of the early onset group of full-term NEC was shock;while those of the late onset group were shock and peritonitis;the risk factors of poor prognosis in the early onset group of preterm NEC were shock and sepsis,while those in the late onset group were pulmonary hemorrhage,shock,intestinal perforation and sepsis.Conclusions Compared to the preterm NEC,the onset time of full-term NEC was earlier and the clinical manifestations were more typical.Early identification and management of shock,peritonitis,intestinal perforation,sepsis and pulmonary hemorrhage can reduce the risk of poor prognosis of neonate NEC.

Objective:To introduce and validate the Pediatric Nausea Assessment Tool (PeNAT) and the Baxter Retching Faces Scale (BARF) in the assessment of chemotherapy induced nausea in Chinese children with malignant neoplasms, and to explore the cut-off value for rescue antiemetic.Methods:A prospective descriptive study was conducted, 244 children in Sun Yat-sen University Cancer Center with malignant neoplasms who received chemotherapy were selected by convenience sampling from July to August 2021. PeNAT, BARF, Visual Analogue Scale (VAS) and the Faces Pain Scale-Revised(FPS-R) were used to assess the severity of nausea and pain before and after chemotherapy, before and 30-60 minutes after the use of rescue antiemetic or analgesic. After chemotherapy, the children also were asked the changes of nausea severity and whether antiemetic was needed.Results:A test-retest reliability was conducted on the patients with the same severity of nausea before and after chemotherapy, and the intraclass correlation coefficient of the PeNAT and BARF were 0.940 (both P<0.05). After chemotherapy, the PeNAT and BARF were 1.5(1.0, 2.0) and 2.0(0, 2.0) points, which were significantly higher than the 1.0(1.0, 1.0) and 0(0, 0) points before chemotherapy ( Z = - 9.19, - 9.09, both P<0.01). The PeNAT and BARF of 11 cases receiving antiemetic before medication were 4.0 (4.0, 6.0) and 3.0(2.0, 4.0) points, which were higher than the 0(0, 2.0) and 1.0(1.0, 2.0) points without antiemetic ( Z = - 4.03, - 3.86, both P<0.05). After chemotherapy, the correlation coefficients between PeNAT or BARF and VAS-nausea were r = 0.933, 0.957 (both P<0.01), and FPS-R were r = 0.192, 0.189 (both P<0.05). After using antiemetic, PeNAT and BARF were 2.0(2.0, 3.0) and 2.5(2.0, 4.0) points, which were significant different than the 3.0(3.0, 3.8) and 4.0(4.0, 8.0) points before using antiemetic ( Z = - 2.97, - 2.83, both P<0.05). According ROC curves and cut-off values, it was determined that PeNAT≥3 and BARF≥4 had clinical significance and require clinical intervention. Conclusions:PeNAT and BARF have excellent reliability and validity in the assessment of chemotherapy induced nausea in children with malignant neoplasms, they can effectively identify the requirement of rescue antiemetic, and evaluate the efficacy of antiemetic.

OBJECTIVETo understand the current situation and characteristics of commercial sexual behavior among old male clients in Guangxi, and to explore the associated risk factors of HIV infection.

METHODSThe areas in Guangxi where more old male HIV cases were reported followed by survey on those male clients who were over 60 years of age, were selected. A total of 5 ml blood sample was collected to test antibodies of HIV and syphilis. Risk factors were calculated by the multivariate logistic analysis method.

RESULTSA total of 1 236 questionnaires were completed in the survey. 65.61% of the old male clients were from low-grade venues with 53.64% of them never using a condom when engaging in commercial sex behavior. 47.33% of them patronized sex workers for around 10 years. 53.32% of them patronized prostitutes not less than 2 times in the last 30 days. Thirty subjects were HIV positive (2.43%) and 40 were syphilis positive (3.24%). Factor as 'having fixed partners' (OR = 0.302, 95% CI:0.140-0.650) appeared protective for the old male clients of HIV infection, with a history of patronizing sex workers from 1 to 5 years (OR = 2.552, 95% CI: 1.086-5.998). Frequency of patronizing sex workers not less than 4 times in the last 30 days (OR = 3.002, 95%CI:1.047-8.607) seemed the high risk factor of HIV infection.

CONCLUSIONOlder-aged male clients showed high HIV and syphilis infection rates in Guangxi, and most of them were using the low-grade venues. Commercial sexual behaviors in the areas would include early and, frequent exposure, but with poor protections. High risk factors of HIV infection might relate to "no fixed sexual partners" and "frequently commercial sexual behavior".

Aged ; Antibodies ; China ; epidemiology ; Condoms ; HIV Infections ; epidemiology ; Humans ; Male ; Middle Aged ; Risk Factors ; Sex Work ; statistics & numerical data ; Sex Workers ; Sexual Behavior ; Sexual Partners ; Surveys and Questionnaires ; Syphilis ; epidemiology