Untargeted metabolomic reveals the changes in muscle metabolites of mice during exercise recovery and the mechanisms of whey protein and whey protein hydrolysate in promoting muscle repair.

Our previous study indicated that whey protein hydrolysate (WPH) showed effective anti-fatigue properties, but its regulatory mechanism on recovery from exercise in mice is unclear. In the present study, we divided the mice into control, WP, and WPH groups and allowed them to rest for 1 h and 24 h after exercise, respectively. The changes in muscle metabolites of mice in the recovery period were investigated using metabolomics techniques. The results showed that the WPH group significantly up-regulated 94 muscle metabolites within 1 h of rest, which was 1.96 and 2.61 times more than the control and WP groups, respectively. In detail, significant decreases in TCA cycle intermediates, lipid metabolites, and carbohydrate metabolites were observed in the control group during exercise recovery. In contrast, administration with WP and WPH enriched more amino acid metabolites within 1 h of rest, which might provide a more comprehensive metabolic environment for muscle repair. Moreover, the WPH group remarkably stimulated the enhancement of lipid, carbohydrate, and vitamin metabolites in the recovery period which might provide raw materials and energy for anabolic reactions. The result of the western blot further demonstrated that WPH could promote muscle repair via activating the Sestrin2/Akt/mTOR/S6K signaling pathway within 1 h of rest. These findings deepen our understanding of the regulatory mechanisms by WPH to promote muscle recovery and may serve as a reference for comprehensive assessments of protein supplements on exercise.

Application of Magnetic Resonance T1rho and T2 Mapping in Evaluating Cartilage Injury in Middle-aged and Elderly Patients with Knee Osteoarthritis.

Objective: To investigate magnetic resonance longitudinal relaxation time quantitative imaging (T1rho) and transverse relaxation time quantitative imaging (T2 mapping) techniques in evaluating cartilage damage in middle-aged and elderly patients with knee osteoarthritis (OA). Methods: To carry out this investigation, the researchers enrolled 65 OA patients subjects for the study. These patients were divided into 2 groups based on the severity of their OA. Thirty healthy individuals were included as the control group. All study participants underwent magnetic resonance T1rho and T2 mapping scans. OA patient scores and values from the Western Ontario and McMaster University Osteoarthritis Index (WOMAC), T2, and a T1rho MRI measurement indicating potential early indication of bone and joint diseases from each cartilage area were compared among the OA patients as well as the control group. Pearson correlation analysis was used to examine the relationships between T2 and T1rho values and WOMAC scores. Results: The WOMAC scores in the mild OA group were lower than the severe OA group (P < .05). There were no significant differences in T2 and T1rho values of lateral tibial cartilage among the 3 groups (P > .05). On the other hand, the T2 and T1rho values of medial femoral, lateral femoral, and medial tibial cartilage areas increased progressively in the control, mild OA, and severe OA groups (P < .05). A Pearson analysis found a positive correlation between the T2 values of medial, lateral, and medial tibial cartilages and the WOMAC scores. Similarly, the T1rho values of these cartilage areas were also positively correlated with the WOMAC scores. Conclusion: Magnetic resonance T1rho and T2 mapping offer good evaluation value for assessing cartilage injury in middle-aged and elderly patients with knee OA. The values obtained from T1rho and T2 mapping in various areas of the cartilage show a positive correlation with WOMAC scores.

The Degree of Hydrolysis and Peptide Profile Affect the Anti-Fatigue Activities of Whey Protein Hydrolysates in Promoting Energy Metabolism in Exercise Mice.

The purpose of this study was to investigate the effects of characteristics of whey protein hydrolysates (WPHs) on energy metabolism in exercise mice. Results showed that high-degree of hydrolysis (DH) hydrolysates (22%, H-Alc and H-AXH) showed better anti-fatigue effects than low-DH hydrolysates (10%, L-Alc and L-AXH) in enhancing energy substances and reducing metabolic byproducts. It might be related to the higher content of components less than 3 kDa in H-Alc and H-AXH (92.35 and 81.05%, respectively) and higher intensities of small peptides containing two to nine residues. Moreover, Western blot results revealed that WPHs maintained the energy balance in exercise mice by regulating the AMP-activated protein kinase (AMPK) and mTOR signaling pathways. Notably, H-Alc had higher intensities of peptides containing two to five residues than H-AXH and these peptides were rich in essential amino acids, which might explain why H-Alc exhibited better effects in decreasing protein metabolites. Meanwhile, H-AXH contained more free amino acids, especially Leu, which might contribute to its ability to promote glucose consumption in muscle. Furthermore, 40 peptides with two to nine residues and high intensities (>5 × 105) were screened from H-Alc and H-AXH and predicted by bioinformatics tools. Among them, LLL, LLF, GTW, AGTW, and ALPM showed high bioavailability, cell permeability, and potential bioactivity.

NOD2 is involved in regulating odontogenic differentiation of DPSCs suppressed by MDP through NF-κB/p65 signaling.

Dental pulp stem cells (DPSCs) are well known for their capable of both self-renewal and multilineage differentiation. Dental tissue diseases, include caries, are often accompanied by inflammatory microenvironment, and muramyl dipeptide (MDP) is involved in the inflammatory stimuli to influence the differentiation of DPSCs. Nucleotide-binding oligomerization domain 2 (NOD2), a member of the cytosolic Nod-like receptor (NLR) family, plays a key role in inflammatory homeostasis regulation, but the role of NOD2 in DPSCs differentiation under inflammatory is still unclear. In this study, we identified that MDP suppressed odontogenic differentiation of DPSCs via NOD2/ NF-κB/p65 signaling pathway. Alizarin red staining and ALP activity showed the odontogenic differentiation was suppressed by MDP in a concentration-dependent manner, and the expression of dentin differentiation marker protein dentin matrix protein 1 (DMP-1) and dentin Sialophosphoprotein (DSPP) also indicated the same results. The expression of NOD2 increased gradually with the concentration of MDP as well as the phosphorylation and nuclear translocation of p65, which meant NF-κB signaling pathway was activated. Further, the interference of NOD2 inhibited the phosphorylation and nuclear translocation of p65 and reversed the MDP-mediated decrease of odontoblast differentiation of DPSCs. Our study showed that MDP can inhibit the odontoblast differentiation of DPSCs in a concentration-dependent manner. The NF-κB signaling pathway was activated by increasing expression of NOD2. Interference of NOD2 reversed the negative ability odontoblast differentiation of DPSCs in the inflammatory environment. Our study might provide a theoretical basis for the clinical treatment for dentinogenesis of DPSCs.

[Ultrasound anatomy and needle-knife insertion approach of common tendon lesions in knee osteoarthritis based on meridian sinew theory].

OBJECTIVE: To explore the ultrasonic anatomical characteristics and needle-knife insertion approach of common tendon lesions in knee osteoarthritis (KOA), so as to provide the references for accurate release of KOA by needle- knife along tendon lesions based on meridian sinew theory. METHODS: Sixty patients with one-knee KOA were selected. High-frequency musculoskeletal ultrasound was used to collect sonograms at the anatomical positions of "Hedingci" "Binwaixia" "Binneixia" and "Yinlingshang". The anatomic levels were marked on the sonograms. The anatomic levels and sonographic features of lesions were compared and analyzed, and the relevant data of needle-knife simulation approach was measured. RESULTS: The "Hedingci" lesions were mainly located at the attachment of quadriceps tendon to patella and suprapatellar bursa. The "Binwaixia" and "Binneixia" lesions were mainly located at the attachment of retinaculum patellae laterale and retinaculum patellae mediale to patella and infrapatellar fat pad. The "Yinlingshang" lesions were mainly located at the attachment of goose foot tendon to medial tibial condyle and bursa of goose foot. With "Hedingci" as an example, when the needle-knife entry point was 1 cm above the patella, the attachment of quadriceps tendon to patella was released, and the average depth of needle-knife was (3.60±0.10) cm, and the needle body was perpendicular to the skin. The average depth of needle-knife for releasing suprapatellar bursa was (2.35±0.17) cm, and the needle body was 45° towards head. CONCLUSION: The musculoskeletal ultrasound could clearly show the local detailed anatomical level, ultrasonic characteristics and anatomical level of common tendon lesions of KOA, and could improve the accuracy of needle-knife along tendon lesions with non-direct vision, which has important reference value for needle-knife medical standardization and standardized operation.

The Transmission and Evolution of HIV-1 Quasispecies within One Couple: a Follow-up Study based on Next-Generation Sequencing.

Next-generation sequencing (NGS) has been successfully used to trace HIV-1 infection. In this study, we investigated the transmission and evolution of HIV-1 quasispecies in a couple infected through heterosexual behavior. A heterosexual couple in which both partners were infected with HIV-1 was followed up for 54 months. Blood samples including whole-blood and plasma samples, were collected at various time points. After HIV-1 subtyping, NGS (Miseq platform) was used to sequence the env region of the HIV-1 quasispecies. Genetic distances were calculated, and phylogenetic trees were generated. We found both partners were infected with HIV-1 subtype circulating recombinant form (CRF), CRF65_cpx. The quasispecies distribution was relatively tightly clustered in the phylogenetic tree during early infection. Over time, the distribution of HIV-1 quasispecies gradually became more dispersed at 12th months, with a progressive increase in gene diversity. By 37th months, the sequences obtained for both partners formed different clusters in the phylogenetic tree. These results suggest that the HIV-1 contact tracing results generated by the Miseq platform may be more reliable than other conventional sequencing methods, which can provide important information about the transmission and evolution of HIV-1. Our findings may help to better target preventative interventions for promoting public health.

Upregulation of PAG1/Cbp contributes to adipose-derived mesenchymal stem cells promoted tumor progression and chemoresistance in breast cancer.

C-terminal Src kinase (Csk)-binding protein (Cbp) is a ubiquitously expressed transmembrane adaptor protein which regulating Src family kinase (SFK) activities. Although SFKs are well known for their involvement in breast cancer, the function of Cbp in breast carcinogenesis upon the adipose-tumor microenvironment has not been investigated. Here, we reported that adipose-derived mesenchymal stem cells (ASCs) induced increased expression of Cbp accompanied by enhanced cell proliferation and chemotherapy resistance in breast cancer cell MCF-7/ADR. Depletion of Cbp in breast cancer cell by RNA interference led to remarkable inhibition of cell proliferation, invasion as well as synergy with adriamycin hydrochloride to suppress the tumor growth. Furthermore, silencing of Cbp concomitantly inhibited the expression of phosphoryl of Src, AKT and mTOR signals. Our study highlights the underlying mechanism of cross interaction between ASCs and breast cancer cells, and indicates that PAG1/Cbp in breast cancer cell may modulate tumor progression and acquired chemoresistance in the ASCs-associated breast cancer microenvironment through Src and AKT/mTOR pathways.

Overexpression of NOTCH-regulated Ankyrin Repeat Protein is associated with papillary thyroid carcinoma progression.

Papillary thyroid cancer (PTC) is one of the endocrine cancers with high clinical and genetic heterogeneity. NOTCH signaling and its downstream NOTCH-Regulated Ankyrin Repeat Protein (NRARP) have been implicated in oncogenesis of many cancers, but the roles in PTCs are less studied. In this study, we show that NRARP is frequently over-expressed in thyroid carcinoma. The over-activation of NRARP is highly and positively correlated with NOTCH genes. Moreover, we find that the expression of NRARP is highly associated with several epithelial mesenchymal transition (EMT) markers and contributes to poor survival outcomes. Therefore, these results indicate that NRARP is an important clinical biomarker in thyroid carcinoma and it promotes EMT induction as well as the progression of PTCs via NOTCH signaling activation.

Hypoxia promotes thyroid differentiation of native murine induced pluripotent stem cells.

Hypothyroidism is a very common hormonal deficiency and the stem cell technology which developed in the recent years may offer a therapeutic strategy for treating this disorder. Hypoxia has been demonstrated to play an important role in embryonic formation and development and to modulate stem cell differentiation. However, the influence of oxygen tension on thyroid differentiation has not been studied. In this study, we used murine induced pluripotent stem (iPS) cells for thyroid cell differentiation under normoxic and hypoxic conditions and compared differentiation efficiency in morphology, function, gene and protein expression under both conditions. We found that hypoxia promoted adhesion and outgrowth of embryoid bodies (EBs) derived from murine iPS cells. Expression of endodermal markers (Foxa2 and Gata4) and thyroid transcription factors (Pax8 and Nkx2.1) was increased by hypoxia at both gene and protein levels during early-mid differentiation stages (p<0.05). And so were the thyroid specific markers NIS and TSHR at the end of the experiment (p<0.05). In addition, functional iodide uptake by differentiated cells was also increased after hypoxia. Thyroid differentiation from iPS cells is enhanced under hypoxia and this may involve hypoxia inducible factors (HIFs) and their downstream gene FGF2. Our data offer a foundation for understanding thyroid development and provide a potentially more efficient way to use cell therapy for treating thyroid deficiency.

[Study on the mortality and risk factors among HIV-infected individuals during 1989 - 2011 in Dehong prefecture, Yunnan province].

OBJECTIVE: To examine the mortality and risk factors among HIV-infected patients during 1989 - 2011 in Dehong prefecture, Yunnan province. METHODS: All HIV-infected patients reported during 1989 - 2011 in Dehong prefecture who held local residency were included in the study. Mortality rates and cumulative survival rates were calculated. Multiple regression analysis under Cox proportional hazard model was conducted to examine the risk factors for deaths. RESULTS: A total of 13 006 HIV-infected patients were included in this study including 73.2% males, 79.1% peasants and 48.7% married at the time of reporting. 64.5% of the patients were ethnic minorities, and 68.7% were illiterate or having received only primary school education. All the patients were followed-up for a total of 55 962.30 person-years with 4648 patients died, with overall mortality rate as 8.31/100 person-years. The mortality rate had been increasing from 1990 to 2004 but decreasing since 2005. The average survival time since the identification of HIV infection was 9.48 years overall, and was 16.65 years for those having received antiretroviral treatment (ART) and 7.67 years for those without ART. Data from multiple regression analysis indicated that ART and socio-demographic characteristics such as age, gender, ethnicity, occupation, marital status, education background etc. were significantly associated with death among HIV-infected patients. CONCLUSION: The comprehensive AIDS campaigns including ART had significantly reduced the deaths among HIV-infected patients in Dehong prefecture. More efforts on the scaling up program of ART as well as the enhanced management and follow-up program tailored for HIV-infected patients with different socio-demographic characteristics were needed to further reduce the deaths in the area.

[Adherence and related determinants on methadone maintenance treatment among heroin addicts in Dehong prefecture, Yunnan province].

OBJECTIVE: To determine the adherence to and its determinants of methadone maintenance treatment (MMT) among heroin addicts in Dehong prefecture in Yunnan province from 2005 to 2009. METHODS: A dynamic cohort analysis was conducted with the time of treatment initiation as the time of cohort entry, to calculate the proportion of adhering to the treatment at different time points after initiation of the treatment, and to study the determinants of adherence. RESULTS: A total of 3758 had been treated. Among them, 95.8% were males, 75.8% aged between 20 - 39 years, 90.4% were peasants or unemployed, 57.0% were ethnic minorities, 35.0% were single and 55.5% were married with spouses, 43.9% were illiterate or educated at most primary school. The age of first using drugs averaged at 23.93 years. About 96.3% of the study subjects had used heroin and 21.7% were HIV-infected. The minimum time under MMT was less than 1 month and the maximum 61 months. The median methadone dose at first delivery was 25 ml, with the minimum 1 ml and the maximum 330 ml. By the end of the study or observation period, a total of 1798 patients had withdrawn from treatment and 1960 were still under treatment. The proportions of adherence to or still being under the treatment after 1, 3, 6, 9, 12, 24, 36, 48 and 60 months treatment were 0.919, 0.847, 0.756, 0.690, 0.637, 0.519, 0.417, 0.360 and 0.321, respectively. Multiple regression analysis using Cox proportional hazard model indicated that withdraw from the methadone maintenance treatment was significantly associated with location of the treatment clinics, year of treatment initiation, marital status, HIV infection status, methadone dose of first delivery and the result of last urine test for heroin use. CONCLUSION: MMT attendants in Dehong prefecture had a relatively high withdraw rate and low adherence rate. More efforts are needed to provide tailored counseling and education to MMT attendants, to provide family and community support, appropriate methadone dose at first delivery, and to better coordinate with local police department.

[Studies on the determinants and changes related to the natural CD4(+) T cell counts among antiretroviral treatment-naive HIV/AIDS patients in Dehong prefecture, Yunnan province].

OBJECTIVE: To explore the determinants and changes of CD4(+) T cell counts among antiretroviral treatment-naive HIV/AIDS patients in Dehong prefecture, Yunnan province. METHODS: A retrospective cohort analysis was conducted on HIV-infected local residents, being reported during 1989 through May 2010, in Dehong prefecture. The patients had received at least two CD4(+) T cell counting tests before receiving the antiretroviral treatment (ART). Difference between the first and the last CD4(+) T cell counts was calculated and described. Logistic regression analysis was conducted to examine the determinants of significant depletion of CD4(+) T cell counts among them. RESULTS: A total of 4487 HIV/AIDS patients were included in the study. The change of CD4(+) T cell counts between the first and the last CD4(+) T cell count tests had a median of -2.0 cells/µl in month (IQR: -8.2 - 3.6) and was significantly associated with socio-demographic characteristics, HIV transmission mode, the first or baseline CD4(+) T cell counts and the time interval between the first and the last CD4(+) T cell counting tests etc. About 60.0% (2693/4487) of the HIV/AIDS patients had deletions of CD4(+) T cell counts, and 31.2% (1400/4487) had significant (≥ 30%) deletions of CD4(+) T cell counts. Results from the multiple logistic regression analysis indicated that age, ethnicity, marital status, HIV transmission mode, the first CD4(+) T cell counts and the interval between the first and the last CD4(+) T cell counting tests were significantly associated with the significant depletion of CD4(+) T cell counts. CONCLUSION: The changing rate of CD4(+) T cell count among ART-naive local HIV-infected patients in Dehong prefecture, Yunnan province was relatively slow. However, substantial proportion of them showed significant decreases of CD4(+) T cell counts, which was determined by many factors. More efforts were needed to systematically and consistently follow-up those HIV-infected patients and measure their CD4(+) T cell counts in China, in order to instantaneously monitor the disease progression, and the initiation of ART, if necessary.

[Causes of death among HIV-infected patients in Dehong prefecture, Yunnan province: 1989-2010].

OBJECTIVES: To analyze the fatality and causes of death related to comprehensive prevention and care programs among HIV-infected patients in Dehong prefecture of Yunnan province, from 1989 to 2010. METHODS: Data on HIV/AIDS death cases in Dehong prefecture were extracted from the "Chinese National Comprehensive HIV/AIDS Prevention and Care Information System" and were analyzed. RESULTS: From 1989 to the end of 2010, a total of 13 493 HIV/AIDS cases registered as local residents or currently living in Dehong, had been reported. Among them, 8569 were reported as HIV cases with 2036 deaths and the other 4924 were reported as AIDS cases with 2251 deaths. A few of the cases had survived for 15 - 20 years. By the end of 2010, the number of deaths was higher than the number of survivors among HIV/AIDS cases reported before 2004, whereas the number of survivors was higher than the number of deaths among HIV/AIDS cases reported in 2004 and there after. During the twenty years' period, the proportion of reported HIV/AIDS cases died in the same year showed a secular trend of being low-highest-low, rising up to > 10.0% in 2001, peaking at 18.9% in 2003 and then continuously going down to 5.8% in 2010. The proportion of HIV/AIDS cases who survived at the beginning but died later in the year was going down since 2007. The proportion of HIV/AIDS deaths died directly from AIDS was increasing whereas the proportion of HIV/AIDS deaths dying directly from overuse of drugs was decreasing in the recent years. Among HIV/AIDS deaths, the proportion of ever received CD4(+) T-cell testing and the proportion of ever having received antiretroviral treatment were also increasing in the past years, reaching to 89.9% and 25.5% in 2010, respectively. CONCLUSION: The case fatality of HIV/AIDS was decreasing in the past years in Dehong prefecture. More efforts were needed to scale up the CD4(+) T-cell count testing and antiretroviral treatment in order to further reduce both morbidity and mortality among HIV/AIDS patients in Dehong prefecture. It is critical to improve surveillance program on HIV/AIDS deaths in the rural areas.

[Proportions and correlates of recent HIV infections among newly reported HIV/AIDS cases from 2005 through 2009 in Dehong prefecture, Yunnan province].

OBJECTIVE: To determine the proportions and correlates of recent HIV infections among newly reported HIV/AIDS cases from 2005 through 2009 in Dehong prefecture, Yunnan province. METHODS: All available serum samples of newly reported HIV/AIDS cases during 2005 - 2009 period in Dehong prefecture, were tested using the BED HIV incidence capture enzyme immunoassay (BED-CEIA). RESULTS: A total of 9367 HIV/AIDS cases were newly reported in 2005 through 2009, of whom 7252 (77.4%) were tested with BED-CEIA. Among the tested, 954 (13.2%) were positive for BED-CEIA and were regarded as recent HIV infections. The proportion of recent HIV infection among newly reported HIV/AIDS cases was 11.21% in 2005, 11.87% in 2006, 17.55% in 2007, 13.22% in 2008, and 12.22% in 2009. Multiple logistic regression analysis indicated that the proportion of recent HIV infections among newly reported HIV/AIDS cases in 2009 was significantly higher among females, those aged 11 - 19 years, and internal residents outside of Dehong prefecture, but significantly lower among immigrants who were mostly from Myanmar, than local residents. CONCLUSION: From 2005 to 2009, the proportion of recent HIV infections among newly reported HIV/AIDS cases in Dehong prefecture in Yunnan province was fluctuating slightly. Future research is needed to examine its long-term and secular trend. Such proportion was significantly different by different sociodemographic characteristics.

[Natural history of HIV infections among injecting drug users in Dehong prefecture, Yunnan province].

OBJECTIVE: To study the natural history of HIV-1 infection among intravenous drug users (IDUs) detected in late 1989 in the study area and the factors related to survival of these IDUs infected with HIV. METHODS: 196 injecting drug users first detected during August and December, 1989 were observed in Ruili county, Yunnan province. Data gathered from the 20-year follow-up program was collected and analyzed retrospectively. RESULTS: After 20 years' follow-up period, 90.3% of the 196 IDUs with HIV infection died, 5.1% of them were still alive, and 4.6% were lost. The crude pre-AIDS mortality rate was 98.1/1000 person-years, and the AIDS mortality rate was 54.9/1000 person-years. Malaria, septicemia were the main causes of death among the natural diseases whereas overdose and accidental causes were the principal causes related to those non-disease deaths. The median survival time from sero-conversion to death was 8.6 years (95%CI: 7.6 - 9.7). The median survival time from sero-conversion to death due to AIDS was 11.3 years (95%CI: 10.3 - 12.8) with the incubation time as around 10.3 years. People older than 30 years at seroconversion and length of drug usage were associated with shorter survival time, with hazards ratios as 1.9 and 0.7, respectively. CONCLUSION: A high pre-AIDS mortality was observed among IDUs. Both the median survival time from sero-conversion to death and the HIV incubation period were shorter than that observed in the developed countries. Age of HIV infection seemed to have a strong effect on survival.

[Tracing the sources of newly reported HIV infections in Dehong prefecture of Yunnan province].

OBJECTIVE: To trace and provide HIV-testing among those having contacts with HIV-infected individuals at various levels in Dehong prefecture, Yunnan province and to evaluate the effectiveness and feasibility of such investigation as a supplemental strategy for HIV testing and control. METHODS: Newly reported HIV infections from August throughout October in Dehong prefecture, in 2008 were asked to provide contact information of persons whom they had high risk contacts with. Persons having had risk contacts with HIV-infected cases were here of interviewed and their blood tested on the sero-status of HIV. RESULTS: A total of 335 HIV cases were newly reported during this three-month period. A total of 309 cases of them and 148 HIV infections identified thereafter from their risk contacts were under informed consent, to participate in this study. A total number of 3395 risk contacts were reported, of whom only 20.7% (704/3395) had 'contact information' and 51.3% (361/704) were successfully located and interviewed, including 117 previously confirmed HIV infections and 244 people with unknown HIV status. The majority of them (203 or 83.2% of 244) were then tested for HIV and 56 (27.6% of 203) were tested positive for HIV. The proportion of having detailed contact information and the proportion of being traced or followed among reported risk contacts of HIV infections were 68.8% and 68.2% for spouses of HIV patients, respectively, which were much higher than those among commercial sex partners (1.2% and 16.7%), casual sex partners (37.3% and 22.3%) and peers who sharing needles (34.1% and 56.4%). CONCLUSION: Newly reported HIV infections reported a large number of risk contacts and new HIV infections were identified among them. It was extremely difficult to trace commercial sex partners or casual sex partners on their HIV infection status. Nevertheless, tracing the risk contacts of newly reported HIV infections seemed to be helpful in identifying new HIV infections and in understanding the nature of transmission towards controlling the HIV epidemics.