Three-parent test tube baby technology is important to solve the mitochondrial genetic disease.Once available,it raises greatly ethical controversy such as breaking traditional family values,hitting the religious belief,existing unknown risks,correctly handling the failed embryo,as well as the influence on the social status of the babies.Regarding these controversy,we can discuss it from several aspects.Because the development of ethics is behind the progress of science and technology,we should affirm the value of three-parent test tube baby technology and balance the development of science and technology with respecting the religious beliefs.Strict supervision system and standard application system reflect our respect for life.Incomprehension to the unknown things should become the motivation of our inquiry.We should face up to our fear of three-parent test tube baby technology,and thus to strengthen research and deepen understanding.Based on the above argument,this paper puts forward the ethical principles that should be followed in the development of three-parent test tube baby technology,namely respect,benefit,no harm and justice.

Since CRISPR/Cas9 has been discovered,it opens a new era for gene therapy with its low cast,high efficiency,short cycle,easy operation and other characteristics.In 2015,Huang Junjiu had used the technique to nodify human embryonic cell for the first time,leading to widespread attention at home and abroad.For nearly 50 years,ethical discussion of gene therapy has never stopped.Through reviewing the development of gene therapy,this paper analyzed several key issues of ethical controversy in gene therapy,such as safety,effectiveness,justice,right,interest orientation and so on,and put forward that gene therapy and ethics are not two opposite sides and cannot be treated separately,the development of gene therapy should follow certain ethical norms and technical norms and it should use the bioethical principles to direct gene therapy.

To investigate the effects of 2-(4-methoxycarbonyl-2-tetradecyloxyphenyl)carbamoylbenzoic acid (CX09040) on protecting pancreatic β cells, the β cell dysfunction model mice were induced by injection of alloxan into the caudal vein of ICR mice, and were treated with compound CX09040. Liraglutide was used as the positive control drug. The amount and the size of islets observed in pathological sections were calculated to evaluate the β cell mass; the glucose stimulated insulin secretion (GSIS) test was applied to estimate the β cell secretary function; the oral glucose tolerance test (OGTT) was taken to observe the glucose metabolism in mice; the expressions of protein in pancreas were detected by Western blotting. The effects on the target protein tyrosine phosphatase 1B (PTP1B) were assessed by the PTP1B activities of both recombinant protein and the intracellular enzyme, and by the PTP1B expression in the pancreas of mice, separately. As the results, with the treatment of CX09040 in alloxan-induced β cell dysfunction mice, the islet amount (P<0.05) and size (P<0.05) increased significantly, the changes of serum insulin in GSIS (P<0.01) and the values of acute insulin response (AIR, P<0.01) were enhanced, compared to those in model group; the impaired glucose tolerance was also ameliorated by CX09040 with the decrease of the values of area under curve (AUC, P<0.01). The activation of the signaling pathways related to β cell proliferation was enhanced by increasing the levels of p-Akt/Akt (P<0.01), p-FoxO1/FoxOl (P<0.001) and PDX-1 (P<0.01). The effects of CX09040 on PTP1B were observed by inhibiting the recombinant hPTP1B activity with IC50 value of 2.78x 10(-7) mol.L-1, reducing the intracellular PTP1B activity of 72.8% (P<0.001), suppressing the PTP1B expression (P<0.001) and up-regulating p-IRβ/IRβ (P<0.01) in pancreas of the β cell dysfunction mice, separately. In conclusion, compound CX09040 showed significant protection effects against the dysfunction of β cell of mice by enlarging the pancreatic β cell mass and increasing the glucose-induced insulin secretion; its major mechanism may be the inhibition on target PTP1B and the succedent up-regulation of β cell proliferation.
Alloxan ; Animals ; Benzoates ; pharmacology ; Biological Assay ; Disease Models, Animal ; Glucose ; metabolism ; Glucose Tolerance Test ; Insulin ; secretion ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; Liraglutide ; pharmacology ; Mice ; Mice, Inbred ICR ; Molecular Weight ; Pancreas ; drug effects ; enzymology ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; antagonists & inhibitors ; Signal Transduction

OBJECTIVETo investigate the effect of allogeneic leukocyte immunization combined with in vitro fertilization-embryo transfer (IVF-ET) for treatment of infertility induced by habitual abortion.

METHODSAllogeneic leukocyte immunization was performed in 9 patients with infertility induced by habitual abortion, with another 9 patients undergoing IVF-ET without habitual abortion as the control group. All the patients were treated with long GnRH-a protocols. The infertility patients with recurrent spontaneous abortion history were immunized with lymphocytes from the husband for before IVF-ET and after clinical pregnancy.

RESULTSThe fertilization rates of the immunotherapy group and control group were 81.3% and 82.2%, respectively, showing no significant difference (P>0.05). Five patients in each group had clinical pregnancy, and a twin pregnancy occurred in the control group. The embryo implantation rates were also comparable between the two groups (22.7% vs 28.6%, P>0.05). All the fetuses resulted from IVF-ET developed normally and were healthily delivered.

CONCLUSIONAllogeneic leukocyte immunotherapy along with IVF-ET is effective for treatment of infertility resulting from recurrent spontaneous abortion.

Abortion, Habitual ; physiopathology ; Adoptive Transfer ; methods ; Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Female ; physiopathology ; therapy ; Pregnancy ; Pregnancy Outcome ; Treatment Outcome

OBJECTIVETo discuss the probable causes of the post-surgery complications with the intra-artcular fracture of calcaneus, the proper steps for prevention and solution.

METHODSSeventy-one patients (76 injured feet) included 49 males and 22 females aged from 19 to 56 years old (mean 35.6 years). According to Sanders' classification, 23 cases (24 injuried feet) belonged to type II, 36 (38 injured feet) were type III, the remain 12 (14 injured feet) met the criteria of type IV. All the patients received the operation of open reduction, autogenous bone grafting and internal fixation with stainless steel plates.

RESULTSThirteen injuried feet developed early complications. Two injuried feet got the superficial layer of the wound disrupted and infected, I had the deep layer of the wound disrupted and infected. Cutaneous necrosis at the pointed end of the wound occurred in 7 cases. Another 1 developed osteomyelitis. Two cases suffered from sural nerve damage. Two injuried feet developed late complications, both of them suffered from arthritis of talocalcaneal joint. All the patients were followed up at least 6 months (ranged from 6 to 42 months, mean 19 months). According to Kerr's post-surgery evaluation criteria, 34 injuried feet were excellent, 32 were fine, 9 were acceptable, only 1 was bad.

CONCLUSIONIf proper measures are taken, the post-surgery complications of intra-articular fracture of calcaneus will be reduced. This requires us to be strict in selecting operation indication, to make a good plan and preparation, to select a right time for operation, to improve surgical skills and pay more attention to peri-surgery nursing. If complications happen, according measures should be taken in order to get a better outcome.

Adult ; Calcaneus ; injuries ; Female ; Follow-Up Studies ; Foot Joints ; injuries ; Fractures, Bone ; pathology ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; prevention & control ; Young Adult

Objective:To construct an outpatient counseling record sheet of breastfeeding based on Delphi method, so as to standardize the consultation process of breastfeeding clinics.Methods:Through literature analysis and discussion of the research group, the expert letter questionnaire was designed by themselves. From September to October 2020, two rounds of Delphi letter inquiries were conducted with 16 experts in Hangzhou to determine the outpatient counseling record sheet of breastfeeding.Results:The positive coefficients of two rounds of expert letter inquiries were 100%, the expert authority coefficients were respectively 0.93 and 0.95, and the Kendall's coefficient of concordance were 0.135-0.204 and 0.234-0.563 ( P< 0.05) . The final counseling record sheet constructed included 8 first-level items, 99 second-level items and 94 third-level items. Conclusions:Letter consulting experts have high enthusiasm, high degree of authority and good coordination. The breastfeeding clinic consultation record based on the Delphi method is highly scientific and practical, which can be applied to breastfeeding clinics in various maternal and infant medical institutions.

Objective To analyze the clinical characteristics and the interventional effects of hysteria in leftbehind children in Chongqing rural area for early detection,early treatment and prevention of the disease.Methods One hundred and three left-behind children aged from 7 to 17 years were diagnosed as hysteria from Jun.2008 to Jun.2011.Their general information,clinical presentations were analyzed retrospectively.After comprehensive intervention,they were assessed the clinical efficacy in accordance with Brief Psychiatric Rating Scale,and some of them were tested by Eysenck Personality Questionnaire (EPQ) and Symptom Checklist (SCL-90).Results There were many clinical forms of hysteria in 103 left-behind children.Physical dysfunction accounted for 72 cases(69.90％),whose symptoms were mainly paralysis and convulsive seizure,which was significantly higher than mental disorder(5 cases,4.9％).The mixed episode of physical dysfunction and mental disorder accounted for 26 cases (25.40％).Seventy-eight cases (75.73 ％) were cured,19 cases (18.45％) improved significantly,and 6 cases (5.83％) improved respectively after intervention of 3 to 12 days.Emotional instability accounted for 12 cases(63.16％) in personality factors with EPQ test in 19 cases.Their factors of somatization,interpersonal relationship,depression,anxiety,and hostility with SCL-90 significantly decreased(all P ＜0.05) after intervention of 7 to 12 days.Conclusions The main forms of hysteria in leftbehind children in Chongqing rural area are physical dysfunction and mixed episode,which are paralysis and convulsive seizure.Emotional instability is common in personality factors for them.It can be achieved good results of hysteria in left-behind children to pay attention to the emotional exchange between the children and their parents,early detect,early treat and active prevent the disease.

OBJECTIVETo study the expression of recombinant human SCF-TPO fusion protein and its biological function.

METHODSFour primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography.

RESULTSThe high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml.

CONCLUSIONSExpressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.

Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; genetics ; physiology ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Recombinant Fusion Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; genetics ; metabolism ; physiology ; Thrombopoietin ; genetics ; metabolism ; physiology

The purpose of the research is to provide a new standard for matching of HLA three-dimensional structure, and summarize the major permissible mismatch and immunogenic mismatch antigens. The molecular modeling method was used to create HLA molecular structures by Swiss Model Server, and the comparison of the differences among the alleles was done by SPDV software with the function of iterative magic fit. The results were recorded by relative mean square deviation (RMSD, nm). The differences among alleles were scattered below 0.06 nm for HLA-A and -B molecules, and below 0.03 nm for HLA-DRB1 molecules. On the basis of the statistical analysis, when RMSD is greater than 0.04 nm for -A and -B molecules and 0.02 nm for -DRB1 molecules, the difference is meaningful and can be related with graft versus host disease. When RMSD is lower than 0.02 nm for -A and -B molecules and 0.01 nm for -DRB1 molecules, the difference is decided unmeaningful. From the data, the permissible mismatch and immunogenic mismatch alleles within HLA-A, HLA-B and HLA-DRB1 molecules were summarized. Three-dimensional structure matching is a new area in the transplantation field, much research should be done in the future.

To probe into the expression of alpha subunit for IL-6R at both mRNA and protein levels in human leukemic cells and to discuss its implication in targeted treatment for leukemia with recombinant IL-6-PE40 exotoxin fusion protein mediated by IL-6/IL-6R system, semi-quantitative RT-PCR, sequencing and FCM were used to analyze the gene and protein expression levels of alpha subunit for IL-6R in leukemic cells. Our results showed that not only mRNA but also protein of alpha subunit for IL-6R are highly expressed in the myelogenous leukemic cell lines, the relative expression levels of mRNA were KG-1(1.22) > (1.02) > U266(1.00) > U937(0.99) > HL-60(0.76). Among lymphoblastic leukemic cell lines, Raji expressed a certain amount of alpha subunit mRNA (0.77), whereas its alpha subunit protein was not detected. Expression of alpha subunit mRNA and protein were negative in lymphoblastic leukemic cell lines, HuT28 and CEM, and chronic myelocytic leukemic cell line K562. These results correlate with those of FCM highly. Noteworthily, normal human peripheral blood mononuclear cells expressed hardly protein of IL-6R alpha subunit. So this study provides sufficient experimental evidence that the targeted treatment by recombinant IL-6-PE40 can specifically kill leukemic cells highly expressing IL-6R without toxicity to normal hematopoietic cells.
Base Sequence ; DNA, Neoplasm ; analysis ; Gene Expression ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; metabolism ; Molecular Sequence Data ; Protein Subunits ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; Receptors, Interleukin-6 ; biosynthesis ; genetics ; Tumor Cells, Cultured ; U937 Cells