[Efficiency of CNV-seq in detecting fetal DMD gene deletion or duplication in prenatal diagnosis].

Objective: To evaluate the diagnostic efficiency of copy number variation sequencing (CNV-seq) to detect the deletion or duplication of DMD gene in prenatal diagnosis. Methods: A retrospective analysis was carried out on the CNV-seq results of 34 544 fetuses diagnosed in the First People's Hospital of Yunnan Province from January 2018 to July 2023. A total of 156 cases of fetuses were collected, including Group 1:125 cases with family history of Duchenne muscular dystrophy or Becker muscular dystrophy (DMD/BMD), and Group 2:31 cases with no family history but a DMD gene deletion or duplication was detected unexpectedly by CNV-seq. Multiplex ligation-dependent probe amplification (MLPA) was used as a standard method to detect the deletion or duplication. Consistency test was carried out basing on the results of CNV-seq and MLPA of all 156 cases. Results: Comparing to MLPA, CNV-seq had a coincidence rate of 92.3% (144/156) for DMD gene deletion or duplication, with a sensitivity and positive predictive value of 88.2%, with a specificity and negative predictive value of 94.3%, a missed detection rate of 3.8%, and a Kappa value of 0.839. CNV-seq missed 4 cases with deletions and 2 with duplications due to involved fragments less than 100 Kb, among 20 cases of deletions and 6 cases of duplications detected by MLPA in Group 1. In Group 2, the deletions and duplications detected by CNV-seq were 42% (13/31) and 58% (18/31), respectively, in which the percentage of duplication was higher than that in Group 1. Among those 18 cases with duplications, 3 cases with duplication locating in exon 42~67 were likely pathogenic; while 9 cases with duplication covering the 5' or 3' end of the DMD gene, containing exon 1 or 79 and with only one breakpoint within the gene, along with the last 6 cases with duplications locating at chrX: 32650635_32910000 detected only by CNV-seq, which might be judged as variants of uncertain significance. Conclusions: CNV-seq has a good efficiency to detect fetal DMD gene deletion or duplication in prenatal diagnosis, while a further verification test by MLPA is recommended. The duplications on chrX: 32650635_32910000, 5' or 3' end of DMD gene detected by CNV-seq should be carefully verified and assessed because those variants appear to be nonpathogenic polymorphisms.

[Appllication of human acellular dermal matrix in surgical treatment of genitourinary disease].

Humanacellular dermal matrix (HADM) is widely used in the field of burn wound repair and tissue engineering plastic surgery. HADM is manufactored by physical and chemical decellular process to remove the antigenic components that might cause immune rejection in dermis.The extracellular matrix of three-dimensional cell scaffold structure with collagen fibers had been used for wound repair and tissue regeneration, while HADM characterized with low absorption rate after implantation and strong ability to induce angiogenesis in host tissue. Studies reported that after the HADM was implanted into the patient, the host cells, such as fibroblasts and myofibroblasts, as well as lymphocytes, macrophages, granulocytes and mast cells, rapidly infiltrated the graft. The connective tissue and neovascularization were then formed within the HADM three-dimensional cell scaffold, the lymphatic system also appears after vascular reconstruction. Traditional urethral reconstruction using autologous skin flaps has some defects, such as complexity of the technology, risk of necrosis of the skin flaps after transplantation, and failure to achieve functional repair of the urethral epithelium. It has been reported that using HADM to reconstruct the urethra in patients with urethral stricture, hypospadias and bladder-vaginal fistula, showed promising results. Others have reported the experience of using HADM to repair and reconstruct congenital classic bladder exstrophy. HADM has also been used for tissue repair in patients with penile skin defect caused by Fonier's gangrene and hidradenitis suppurativa, and implanted under Bucks' fascia to enlarge the penis. The report of HADM implantation for treating premature ejaculation also deserves attention. Researchers found that HADM implantation can form a tissue barrier between the skin and corpus cavernosum, which can effectively reduce penile sensitivity and treat premature ejaculation. The safety and effectiveness of HADM implantation in the treatment of premature ejaculation need to be further standardized by data from multi-center, large-sample clinical studies. In summary, HADM is the extracellular matrix and three-dimensional cell scaffold of human dermis. As a new type of tissue repair material, new blood vessels are formed actively after implantation, which shows good histocompatibility. HADM has shown increasingly broad application prospects in treatment of genitourinary diseases including penis, urethra and bladder diseases. HADM has also been used in the treatment of premature ejaculation in recent clinical studies, and its long-term safety and efficacy need to be further investigated.

[The effect of birth ball on the outcome of COOK balloon induction].

Objective: To investigate the effect of birth ball on the outcome of COOK balloon induction. Methods: A total of 580 patients were hospitalized in the obstetrics department of Jiaxing Maternal and Child Health Hospital from May 1, 2017 to December 31, 2017. According to the patient's wishes, 290 cases of cervical ripening with birth ball combined with COOK balloon were taken as experimental group, and 290 cases of cervical ripening group with COOK balloon were set as control group. Bishop score improvement, labor time, cesarean section rate, delivery forceps rate, postpartum hemorrhage, and neonatal scores were compared between the two groups of patients. Results: (1)There was no significant difference in the ag, Body Mass Index (BMI), education background, registered residence and gravidity between the two groups (P>0.05). (2)There was no significant difference in Bishop score improvement between the two groups (P>0.05). (3)The labor time in the experimental group (8.4 h±1.5 h)was significantly shorter than that in the control group (10.6 h±2.7 h). The differences were statistically significant (P<0.05). The cesarean section rate (11.03% vs 15.17%), forceps delivery rate (5.52% vs 8.28%), and postpartum hemorrhage (308 ml±9 ml vs 367 ml±16 ml) were significantly lower in the experimental group than in the control group. The differences were statistically significant (P<0.05). Conclusion: The application of the birth ball in the induction of the COOK balloon can shorten the labor time, reduce the cesarean section rate and the forceps delivery rate, and reduce the postpartum hemorrhage, which has certain clinical application value.

Classical Fractals and Quantum Chaos in Ultracold Dipolar Collisions.

We examine a dipolar-gas model to address fundamental issues regarding the correspondence between classical chaos and quantum observations in ultracold dipolar collisions. The theoretical model consists of a short-range Lennard-Jones potential well with an anisotropic, long-range dipole-dipole interaction between two atoms. Both the classical and quantum dynamics are explored for the same Hamiltonian of the system. The classical chaotic scattering is revealed by the fractals developed in the scattering function (defined as the final atom separation as a function of initial conditions), while the quantum chaotic features lead to the repulsion of the eigenphases from the corresponding quantum S matrix. The nearest-eigenphase-spacing statistics have an intermediate behavior between the Poisson and the Wigner-Dyson distributions. The character of the distribution can be controlled by changing an effective Planck constant or the dipole moment. The degree of quantum chaos shows a good correspondence with the overall average of the classical scattering function. The results presented here also provide helpful insights for understanding the role of the inherent dipole-dipole interaction in the currently ongoing experiments on ultracold collisions of highly magnetic atoms.

Identification of lactoferrin and glutamate receptor-interacting protein 1 in bovine cervical mucus: A putative marker for oestrous detection.

Accurate detection of oestrus is important for artificial insemination. The aim of this study was to identify oestrous-specific bovine cervical mucus proteins that could be used to determine the optimal time for artificial insemination. Non-oestrous and controlled internal drug release (CIDR)-induced oestrous-stage mucus proteins were purified and subjected to surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, sodium dodecyl sulphate polyacrylamide gel electrophoresis and MALDI-TOF/TOF. Among differentially expressed proteins, lactoferrin (LF) and glutamate receptor-interacting protein 1 (GRIP1) showed a twofold increase during the CIDR-induced oestrous stage compared to the levels in non-oestrous stage in bovine cervical mucus. The RT-PCR, Western blotting and immunohistochemistry results showed that LF and GRIP1 expression was significantly increased during the oestrous stage in the uterus. This study demonstrated that bovine LF and GRIP1 exist during the oestrous stage, but not during the non-oestrous stage, suggesting that cervical mucus LF and GRIP1 are useful oestrous detection markers in cattle.

Expression of IncRNA UCA1 in ovarian cancer and its clinical significance.

OBJECTIVES: To explore the expression and clinical significance of IncRNA-UCA1 in ovarian cancer. MATERIALS AND METHODS: The ex- pression of IncRNA-UCA1 in 26 ovarian cancer tissue and 16 normal and benign ovarian tissue were detected using qRT-PCR method, and the correlation of expression level with clinicopathological features were analyzed. RESULTS: Higher lncRNA-UCA1 expression level were detected in ovarian cancer tissue than those in normal ovarian tissue (p < 0.05). There were significant correlations between higher expression of IncRNA-UCA1 with tumor staging (p = 0.000), histological grades (p = 0.000), peritoneal effusion (p = 0.001), and lymph node metastasis (p = 0.000), but not with age. CONCLUSION: IncRNA-UCA1 may play a vital role in the metastasis of ovarian cancer and it is expected to be a potential novel biomarker and therapeutic target of ovarian cancer.

[Feasibility study of transplantation of penile corpus cavernosum and major pelvic ganglion in renal subserous region].

OBJECTIVE: To study the feasibility of transplantation of normal rat penile corpus cavernosum and major pelvic ganglion (MPG) into the renal subserous region of a Nu/Nu mouse based on allograft technology. METHODS: Penile corpus cavernosum and MPG, harvested from Sprague-Dawley (SD) rats under sterile condition, were transplanted underneath the kidney capsule of Nu/Nu mice through the microsurgery instruments and surgery microscope. The histopathologic changes and cellular proliferation in the transplanted penile corpus cavernosum and MPG were then analyzed at the end of 1week and 4 weeks after transplantation. Histological staining and immunohistochemical staining were used to evaluate the main outcome measures. RESULTS: After 1 week, the tissue morphology of the transplanted corpus cavernosum underneath the kidney capsule of Nu/Nu mice was consistent with normal penile corpus cavernosum, and blood could be observed in the penis cavernous sinus of the graft; after 4 weeks, the mophorlogy of the tranplanted corpus cavernosum near the kidney was consistent with normal penile corpus cavernosum, while fibrosis was noteworthy in the graft away from the kidney, but blood could still be seen in the penis cavernous sinus. After 1 week, the tissue morphology of the transplanted MPG was consistent with normal MPG, multiple islet-like cell clusters could be seen in the transplanted MPG in the renal subserous region, and angiogenesis could be observed near the kidney; after 4 weeks, a network of blood vessels was clearly visible away from the kidney, and islet-like cell clusters were still clearly observed in the transplanted MPG. In addition, ki67 positive cells were observed in the transplanted penile corpus cavernosum and MPG after 4 weeks of transplantation, which indicated that there was still cell proliferation activity in the grafts. CONCLUSION: The transplanted corpus cavernosum and MPG underneath the kidney capsule of Nu/Nu mice could survive at least 4 weeks. Moreover, the inner structure of the transplanted corpus cavernosum and MPG was close to the normal tissue. The underlining mechanism may be related to the local microenvironment underneath the kidney capsule of Nu/Nu mice and the neovascularization in the transplanted grafts.

[Feasibility study of transplantation of penile corpus cavernosum and major pelvic ganglion in renal subserous region].

OBJECTIVE: To study the feasibility of transplantation of normal rat penile corpus cavernosum and major pelvic ganglion (MPG) into the renal subserous region of a Nu/Nu mouse based on allograft technology. METHODS: Penile corpus cavernosum and MPG, harvested from Sprague-Dawley (SD) rats under sterile condition, were transplanted underneath the kidney capsule of Nu/Nu mice through the microsurgery instruments and surgery microscope. The histopathologic changes and cellular proliferation in the transplanted penile corpus cavernosum and MPG were then analyzed at the end of 1week and 4 weeks after transplantation. Histological staining and immunohistochemical staining were used to evaluate the main outcome measures. RESULTS: After 1 week, the tissue morphology of the transplanted corpus cavernosum underneath the kidney capsule of Nu/Nu mice was consistent with normal penile corpus cavernosum, and blood could be observed in the penis cavernous sinus of the graft; after 4 weeks, the mophorlogy of the tranplanted corpus cavernosum near the kidney was consistent with normal penile corpus cavernosum, while fibrosis was noteworthy in the graft away from the kidney, but blood could still be seen in the penis cavernous sinus. After 1 week, the tissue morphology of the transplanted MPG was consistent with normal MPG, multiple islet-like cell clusters could be seen in the transplanted MPG in the renal subserous region, and angiogenesis could be observed near the kidney; after 4 weeks, a network of blood vessels was clearly visible away from the kidney, and islet-like cell clusters were still clearly observed in the transplanted MPG. In addition, ki67 positive cells were observed in the transplanted penile corpus cavernosum and MPG after 4 weeks of transplantation, which indicated that there was still cell proliferation activity in the grafts. CONCLUSION: The transplanted corpus cavernosum and MPG underneath the kidney capsule of Nu/Nu mice could survive at least 4 weeks. Moreover, the inner structure of the transplanted corpus cavernosum and MPG was close to the normal tissue. The underlining mechanism may be related to the local microenvironment underneath the kidney capsule of Nu/Nu mice and the neovascularization in the transplanted grafts.

[Prenatal diagnosis of chromosome abnormalities and nine microdeletion syndromes using both traditional karyotyping and BoBs].

OBJECTIVE: To evaluate a new prenatal diagnosis model of chromosomal abnormalities and nine microdeletion syndromes by using both traditional karyotyping and a newly-developed rapid prenatal diagnosis technology, BACs-on-Beads (BoBs) technique. METHODS: From June 2012 to December 2014, 807 pregnant women with high risk after screening or with other indicators, were performed amniocentesis. Traditional karyotyping and BoBs were employed simultaneously for prenatal diagnosis. RESULTS: Thirty-two cases with chromosome aneupoidies were successfully detected both by BoBs and karyotyping, including 18 cases of trisomy 21, 6 cases of trisomy 18, 1 case of trisomy 13, and 7 cases with sex chromosome abnormality. All 8 fetuses with chromosome structural abnormalities detected by karyotyping were missed by BoBs; while BoBs contributed more in detection of five microdeletion syndrome cases, including 3 cases of DiGeorge syndromes (two with microduplication and one with microdeletion), one case of Miller-Dieker syndrome, and one case of Wolf-Hirschhorn syndrome. CONCLUSION: Combined use of traditional karyotyping and BoBs, is a rapid and effective prenatal diagnosis model that may enlarge our horizon on chromosomal diseases and should be widely used in future clinical service.

A novel MICA allele, MICA*069, identified by sequence-based typing in a Chinese individual.

MICA*069 differs from MICA*010:01 by one nucleotide at position 1118 within exon 6.

Protein Profile in Corpus Luteum during Pregnancy in Korean Native Cows.

Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism, but the profile of proteins associated with progesterone synthesis in cyclic and pregnant corpus luteum (CL) is not well-known in cattle. In Experiment 1, plasma progesterone level was monitored in cyclic cows (n = 5) and pregnant cows (n = 6; until d-90). A significant decline in the plasma progesterone level occurred at d-19 of cyclic cows. Progesterone level in abbatoir-derived luteal tissues was also determined at d 1 to 5, 6 to 13 and 14 to 20 of cyclic cows, and d-60 and -90 of pregnant cows (n = 5 each). Progesterone level in d-60 CL was not different from those in d 6 to 13 CL and d-90 CL, although the difference between d 6 to 13 and d-90 was significant. In Experiment 2, protein expression pattern in CL at d-90 (n = 4) was compared with that in CL of cyclic cows at d 6 to 13 (n = 5). Significant changes in the level of protein expression were detected in 32 protein spots by two-dimensional polyacrylamide gel electrophoresis (2-DE), and 23 of them were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Six proteins were found only in pregnant CL, while the other 17 proteins were found only in cyclic CL. Among the above 6 proteins, vimentin which is involved in the regulation of post-implantation development was included. Thus, the protein expression pattern in CL was disorientated from cyclic luteal phase to mid pregnancy, and alterations in specific CL protein expression may contribute to the maintenance of pregnancy in Korean native cows.

Characterization of the genomic full-length sequence of HLA-Cw*010201, Cw*0706, and Cw*0801 in Chinese individuals.

Genomic full-length sequence of human leukocyte antigen (HLA)-Cw*0706, differing from its closet allele Cw*070 101 by six nucleotide exchanges, was detected in two Chinese individuals.

Genomic full-length sequence of HLA-Cw*0103 and *0108, identified by cloning and sequencing.

Genomic full-length sequences of human leukocyte antigen (HLA)-Cw*0103 and *0108 were identified by cloning and sequencing from two Chinese donors. All introns, exons 4-8, 5'-promoter, and 3'-UTR were found to be identical between these two alleles.

Conformation changes of albumin and lysozyme on electrospun TiO2 nanofibers and its effects on MSC behaviors.

Protein adsorption plays a key role in bone repair and regeneration by affecting cell behavior. In this study, TiO2 nanofibers (TiO2 NFs) with different structures, including anatase TiO2 nanofibers (A-NFs), anatase TiO2 nanofibers with beads (B-NFs), anatase-rutile TiO2 nanofibers (AR-NFs) and rutile TiO2 nanofibers (R-NFs), were prepared by electrospinning method. Bovine serum albumin (BSA) and lysozyme (LYZ) were used to explore the adsorption behaviors of TiO2 NFs and then the effects of materials with protein on bone marrow mesenchymal stem cells (MSCs) were studied. Pure titanium metal (PT) was used as control. The results displayed that the adsorption amounts of BSA on samples were B-NFs > AR-NFs > A-NFs ≈ R-NFs > PT, and that for LYZ were B-NFs > AR-NFs > R-NFs > A-NFs > PT. The conformation of proteins changed remarkably when they were adsorbed on meterials. Soaking the TiO2 NFs with and without protein into SBF revealed that the BSA and LYZ on B-NFs, A-NFs and AR-NFs could accelerate the HA deposition on its surface, but it had no promoting effect on HA deposition on B-NFs. MTT and PCR tests showed that the BSA and LYZ adsorbed on materials could promote the proliferation and osteogenic differentiation of MSCs to different degrees due to their different adsorption amount and conformation changes on different TiO2 NFs. The current work demonstrated that the surface properties and crystal structure of TiO2 NFs could influence the adsorption behavior and conformational change of BSA and LYZ, and then further regulate MSCs biological behavior.

Time-dependent exposure of nicotine and smoke modulate ultrasubcellular organelle localization of dopamine D1 and D2 receptors in the rat caudate-putamen.

The correlation of the subcellular localization of dopamine D(1) and D(2) receptors (DA D(1) R, DA D(2) R) with nicotine addiction has not been studied. We demonstrated the ultrasubcellular organelle localization of DA D(1) and D(2) Rs in the caudate-putamen (CPu) area of rat brain in vivo exposed to nicotine (3 mg/day; oral) and passive cigarette smoking (500 ml each; 3 times/day) for 1, 4, and 12 weeks, respectively. Our results revealed DA D(1) R localization in the presynaptic and postsynaptic dendrites, endocytic vesicles, and secretory granules, and DA D(2) R localization in the presynaptic dendrites and vesicles. DA D(1) R immunogold particles were highly decreased in the secretory granules of CPu, and increased in the postsynaptic area and vesicles after prolonged nicotine and smoking exposures, suggesting the strong influence of long time smoking and nicotine exposures on DA D(1) R subcellular organelle localization. DA D(2) R immunoreactivity was comparatively less changed than that of the DA D(1) R. Western blot analysis also showed the differential expression of DA D(1) and D(2) R proteins upon nicotine and smoking exposures as compared to the untreated controls. Taken together, the results for the first time suggests the execution of addictive behavior of nicotine through modulation of mesolimbic dopaminergic system targeting subcellular organelle of DA D(1) and D(2) Rs in the CPu of adult rat brain that may lead to novel therapeutic approaches related to nicotine's neuropsychological disorders including drug addiction.

Synergistic effects of fibronectin and bone morphogenetic protein on the bioactivity of titanium metal.

To improve the biological properties of bioactive titanium metal, recombinant human bone morphogenetic protein 2(rhBMP-2) and fibronectin (Fn) were adsorbed on its surface solely or contiguously to modify the anodic oxidized titanium (AO-Ti), acid-alkali-treated titanium (AA-Ti), and polished titanium (P-Ti). It is found that the different bioactive titanium surface structures had great influence on protein adsorption. The adsorption amounts of BMP adsorbed solely and Fn/BMP adsorbed contiguously were AA-Ti > P-Ti > AO-Ti, and that for Fn adsorbed solely was AA-Ti ≈ P-Ti > AO-Ti. The conformation of proteins was changed remarkably after the adsorption. For BMP, the α-helix decreased on AA-Ti and stabilized on P-Ti and AO-Ti. For Fn, the ß-sheet on PT-Ti and AA-Ti increased significantly. For Fn/BMP, the percentage of ß-sheet on AA-Ti increased, and that of α-helix on all samples was stable. MSCs showed greater adhesion and spreading on Fn/BMP groups. MTT and Elisa tests showed that the synergistic effects of proteins made the cells proliferate and differentiate faster. It indicated both the surface structure and the synergistic effects of proteins could influence the biological properties of titanium metals. It provides research foundation for improving the biological properties of bioactive titanium metals by simultaneous application of several proteins. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2485-2498, 2017.

Bio-functionalization of biomedical metals.

Bio-functionalization means to endow biomaterials with bio-functions so as to make the materials or devices more suitable for biomedical applications. Traditionally, because of the excellent mechanical properties, the biomedical metals have been widely used in clinic. However, the utilized functions are basically supporting or fixation especially for the implantable devices. Nowadays, some new functions, including bioactivity, anti-tumor, anti-microbial, and so on, are introduced to biomedical metals. To realize those bio-functions on the metallic biomedical materials, surface modification is the most commonly used method. Surface modification, including physical and chemical methods, is an effective way to alter the surface morphology and composition of biomaterials. It can endow the biomedical metals with new surface properties while still retain the good mechanical properties of the bulk material. Having analyzed the ways of realizing the bio-functionalization, this article briefly summarized the bio-functionalization concepts of six hot spots in this field. They are bioactivity, bony tissue inducing, anti-microbial, anti-tumor, anticoagulation, and drug loading functions.

Preparation of bioactive nanotitania ceramics with biomechanical compatibility.

In this article, bioactive nanotitania ceramics with biomechanical compatibility was prepared by using an additive of hydroxyapatite or MgO as particle growth inhibitor. After sintering at 1000 degrees C, the particle size of nanotitania ceramics prepared by using HA as additive (HT) was much smaller than that prepared by using MgO as additive (MT). In simulated body fluid (SBF), HT could induce apatite formation in 4 days, while no apatite could be found on MT even after it was soaked in SBF for 14 days. After Ros17/28 osteoblasts were cultured on the materials for 1, 4, and 6 days, MTT results showed that the osteoblasts on the HT differentiated faster than that on the MT. Mechanical tests results showed that the bending and compressive strength of HT were 160 and 200 MPa, while those of MT were 70 and 88 MPa, respectively. These results demonstrated that it is suitable to prepare bioactive nanotitania ceramics, with biomechanical compatibility, by using HA as particle growth inhibitor.

The influence of surface bioactivated modification on titanium percutaneous implants anchored in bone.

In order to achieve biological sealing and resist mechanical damage of load-bearing percutaneous devices, Ti with excellent mechanical properties was anodic-oxidized to be endowed with bioactivity, with plasma-sprayed hydroxyapatite coated Ti as control. Similar to previous works, hydroxyapatite coating could bond tightly with living tissues, resulting in implant stability for whole implantation periods. Meanwhile, when anodic-oxidized bioactivated Ti was implanted percutaneously in vivo, it could induce a layer of calcium phosphate at the interface of tissues/implant. This layer of Ca-P not only induced the fibrous tissue or collagen ingrowth in its structure, but also improved the osteointegration between the bone and the implant. There was no significant biological response difference for the anodic-oxidized Ti and HA coated Ti at different implantation period with histological statistical analysis (p>0.05). Accordingly, suitable bioactivated modified surface of Ti by anodic-oxidized method could not only obtain the same results as the HA coating, but also might avoid some drawbacks of plasma-sprayed HA coatings to achieve biological sealing for a long period in vivo.

Transient induction of apoptosis in serum-starved glioma cells by insulin and IGF-1.

Insulin has a wide variety of biological effects. One of them is a mitogen-like activity whereby cell proliferation is stimulated. In this study we found a heretofore unreported insulin-elicited transient apoptosis of glioma cells. When serum-starved glioma cells were fed with a fresh regular medium, in the 6- to 12-h post-starvation period, the growth rate as determined by cell number was significantly suppressed by insulin, although cell cycle progression and DNA synthesis were actually accelerated. Increase in apoptosis in those growth-retarded cultures was demonstrable by Hoechst staining, detection of histone-associated DNA fragment, and in situ cell death detection. Apoptosis occurred among cells in all stages of cell cycle. After 24 h post-starvation, insulin increased the total cell number like a typical growth-promoting mitogen. In this regard, IGF-1, but not EGF nor TGF-beta 1, behaved like insulin.