Changes in the ECM such as accumulation of high molecular weight of HA by HAS and degradation of HA by endogenous HYAL were essential for adipogenesis both in vitro and in vivo.

We read with interest the comments raised by Gong et al.1 on our recently published study, 2 and appreciate these important and critical remarks.We agree that contamination is a potential important problem with polymerase chain reaction (PCR)-based work. To avoid the possibility of contamination, we confirmed our data before completion of our manuscript. We performed the PCR experiment using new experimental reagents at a different laboratory. We also repeated the PCR to detect the fusion using two different set of primers, the new primer set producing longer amplicons than the old primer set. All these efforts produced identical results, confirming that our samples were not contaminated by PCR amplicons.As the direct fusion of one exon of CTLA4 to an exon of CD28 is less likely than fusion breakpoints within the introns, we confirmed direct fusion of the two exons at the genomic DNA level with other experiments before submitting our manuscript. As shown in Figure 1, direct fusion of exon 3 of CTLA4 to exon 4 of CD28 was confirmed by PCR using primers from intron 2 of CTLA4 and the 3' region of CD28 using genomic DNA from formalin-fixed, paraffin-embedded samples. PCR products were validated by Sanger sequencing. DNA from normal blood was used as a control as well as no template controls. These data confirm that the fusion is present at the DNA level and is not the result of contamination.We also tried to perform fluorescent in situ hybridization experiments. However it was difficult to design probes which can distinguish the fusion segment because the CD28 and CTLA4 genes are located very close to each other. As shown in our manuscript 2 in Online Supplementary Figure S5, the result was not helpful for delineating the fusion signal from the normal signal.

Objectives To evaluate efficacy of the antenatal, intra-partum and post-natal antiretroviral components of a public service Prevention of Mother to Child (MTCT) program in infants. Design Analysis of prospectively collected screening data of demographic and MTCT-related interventions and HIV-infection status of infants identified through HIV-specific DNA polymerase chain reaction. Setting Tygerberg Children’s Hospital, Western Cape Province, South Africa. Subjects HIV-infected women and their infants identified through participation in a public service MTCT program were referred for possible participation in a prospective study of isoniazid prophylaxis. Interventions Key components of the Program include voluntary counselling and testing, zidovudine to the mother from between 28 and 34 weeks gestation and to the newborn infant for the first week, single dose nevirapine to the mother in labour and the newborn shortly after birth and free formula for 6 months. Main Outcome Measures Number and percentage of HIV-infected infants and extent of exposure to antenatal, intrapartum and post-natal antiretrovirals. Results Of 656 infants with a median age of 12.6 weeks, screened between April 1st 2005 through May 2006, 39 were HIV-infected giving a transmission rate of 5.9% (95% CI: 4.4% – 8.0%). Antenatal prophylaxis was significantly associated with reduced transmission (OR: 0.43 (95% CI: 0.21 – 0.94)) as opposed to intrapartum and postpartum components (p=0.85 and p=0.84, respectively). In multivariable analysis the antenatal component remained significant (OR=0.40 (95% CI 0.19 – 0.90)). Conclusions The antenatal phase is the most important antiretroviral component of the MTCT program, allowing most opportunity for intervention.