Boosting Stimulated Emission via a Hydrogen-Bonded Co-Crystallization Approach in Molecular Crystals.

Stimulated emission of organic π-conjugated molecule in solid state remains a significant challenge, mainly involving the mode of molecular stacking that invariably alters the photo-physical processes. Herein, we successfully realized the stimulated emission in molecular crystals using a hydrogen-bonded co-crystallization strategy. Two hydrogen-bonded co-crystals, obtained from 1,4-bis-p-cyanostyrylbenzene (CNDSB) and two types of co-formers, can boost stimulated emission and show decent amplified spontaneous emission (ASE), whereas the parent CNDSB crystal is not SE-active. Crystal structural analysis demonstrated that the co-crystallization eliminated excimer formation. The resulting higher kr and shorter excited-lifetime led to a larger stimulated-emission cross section, which benefited to the occurrence of ASE. Simultaneously, the uniaxial arrangements along long axis of co-crystal together contributed to highly polarized emission. This system presents very rare evidence of boosting stimulated emission by binary co-crystallization, which enriches our insights into organic solid-state lasers.

Stress response and tolerance mechanisms of NaHCO3 exposure based on biochemical assays and multi-omics approach in the liver of crucian carp (Carassius auratus).

The development and utilization of saline-alkaline water, an important backup resource, has received widespread attention. However, the underuse of saline-alkaline water, threatened by the single species of saline-alkaline aquaculture, seriously affects the development of the fishery economy. In this work, a 30-day NaHCO3 stress experimental study combined with analyses of untargeted metabolomics, transcriptome, and biochemical approaches was conducted on crucian carp to provide a better understanding of the saline-alkaline stress response mechanism in freshwater fish. This work revealed the relationships among the biochemical parameters, endogenous differentially expressed metabolites (DEMs), and differentially expressed genes (DEGs) in the crucian carp livers. The biochemical analysis showed that NaHCO3 exposure changed the levels of several physiological parameters associated with the liver, including antioxidant enzymes (SOD, CAT, GSH-Px), MDA, AKP, and CPS. According to the metabolomics study, 90 DEMs are involved in various metabolic pathways such as ketone synthesis and degradation metabolism, glycerophospholipid metabolism, arachidonic acid metabolism, and linoleic acid metabolism. In addition, transcriptomics data analysis showed that a total of 301 DEGs were screened between the control group and the high NaHCO3 concentration group, of which 129 up-regulated genes and 172 down-regulated genes. Overall, NaHCO3 exposure could cause lipid metabolism disorders and induce energy metabolism imbalance in the crucian carp liver. Simultaneously, crucian carp might regulate its saline-alkaline resistance mechanism by enhancing the synthesis of glycerophospholipid metabolism, ketone bodies, and degradation metabolism, at the same time increasing the vitality of antioxidant enzymes (SOD, CAT, GSH-Px) and nonspecific immune enzyme (AKP). Herein, all results will provide new insights into the molecular mechanisms underlying the stress responses and tolerance to saline-alkaline exposure in crucian carp.

Solvation-Mediated Self-Assembly from Crystals to Helices of Protic Acyclic Carbene AuI -Enantiomers with Chirality Amplification.

Constructing chiral supramolecular assembly and exploring the underlying mechanism are of great significance in promoting the development of circularly polarized luminescence (CPL)-active materials. Herein, we report a solvation-mediated self-assembly from single-crystals to helical nanofibers based on the first protic acyclic (methoxy)(amino)carbenes (pAMACs) AuI -enantiomers driven by a synergetic aurophilic interactions and H-bonds. Their aggregation-dependent thermally activated delayed fluorescence properties with high quantum yields (ΦFL ) up to 95 % were proved to be attributed to packing modes of Au⋅⋅⋅Au dimers with π-stacking or one-dimensional extended Au⋅⋅⋅Au chains. Via drop-casting method, supramolecular P- or M-helices were prepared. Detailed studies on the helices demonstrate that formations of extended helical Au⋅⋅⋅Au molecular chains amplify supramolecular chirality, leading to strong CPL with high dissymmetry factor (|glum |=0.030, ΦFL =67 %) and high CPL brightness (BCPL ) of 4.87×10-3 . Our findings bring new insights into the fabrication of helical structures to improve CPL performance by modifying aurophilic interactions.

Design and synthesis of fluorinated peptides for analysis of fluorous effects on the interconversion of polyproline helices.

The unique interaction between fluorine atoms has been exploited to alter protein structures and to develop synthetic and analytical applications. To expand such fluorous interaction for novel applications, polyproline peptides represent an excellent molecular nanoscaffold for controlling the presentation of perfluoroalkyl groups on their unique secondary structure. We develop approaches to synthesis fluorinated peptides to systematically investigate how the number, location and types of the fluorous groups on polyproline affect the conformation by monitoring the transition between the two major polyproline structures PPI and PPII. This work provides valuable information on how fluorous interaction affects the peptide structure and also benefits the design of functional fluorous molecules.

Anti-phytopathogenic activity of sporothriolide, a metabolite from endophyte Nodulisporium sp. A21 in Ginkgo biloba.

Phytopathogenic fungi such as Rhizoctonia solani and Sclerotinia sclerotiorum caused multiple plant diseases resulting in severe loss of crop production. Increasing documents endorsed that endophytes are a striking resource pool for numerous metabolites with various bioactivities such as anti-fungal. Here we reported the characterization and anti-phytopathogenic activity of sporothriolide, a metabolite produced by Nodulisporium sp. A21-an endophytic fungus in the leaves of Ginkgo biloba. Among the total twenty-five endophytic fungi isolated from the healthy leaves of G. biloba, the fermentation broth (FB) of the strain A21 was found potently inhibitory activity against R. solani and S. sclerotiorum using mycelia growth inhibition method. A21 was then identified as Nodulisporium sp., the asexual stage of Hypoxylon sp., by microscopic examination and ITS rDNA sequence data comparison. Under the bioassay-guided fractionation, sporothriolide was isolated from the petroleum ether extract of the FB of A21, whose structure was established by integrated interpretation of HR-ESI-MS and (1)H- and (13)C-NMR. Furthermore, the crystal structure of sporothriolide was first reported. In addition, sporothriolide was validated to be potently antifungal against R. solani, S. sclerotiorum and inhibit conidium germination of Magnaporthe oryzae in vitro and in vivo, indicating that it could be used as a lead compound for new fungicide development.

New flavonoid from Patrinia villosa.

CONTEXT: Patrinia villosa (Thunb.) Juss (Valerianaceae) is an important ancient herbal medicine widely used for inflammation, wound healing, and abdominal pain. But little is known of the phytochemical constituents of this herbal plant. OBJECTIVE: The objective of this study is to isolate and identify the bioactive components from P. villosa. MATERIALS AND METHODS: A 70% EtOH extract of P. villosa was subjected to normal-phase silica, ODS silica gel column chromatography, and semi-preparative HPLC chromatography after partitioned successively with light petroleum, dichloromethane and n-BuOH. Chemical structures of the compounds were elucidated by spectroscopic methods including UV, 1D-NMR, 2D-NMR, HR-ESI-MS, and CD spectra. The cytotoxic activity of the new component was determined with the SMMC-7721 cell line using the MTT method after incubation for 48 h. RESULTS: A new flavonoid named patriniaflavanone A (1) along with four known compounds was isolated from P. villosa. The four known compounds were identified as luteolin 7-O-glucuronide-6″-methyl ester (2), p-hydroxyphenylacetic acid methyl ester (3), trans-caffeic acid (4), and trans-caffeic acid methylate (5) by comparison of their spectral data with the reported data. The IC50 value of patriniaflavanone A (1) on SMMC-7721 was 61.27 µM. DISCUSSION AND CONCLUSION: This is the first report on the isolation and identification of patriniaflavanone A (1), and compounds 2-5 were isolated for the first time from the title plant. Patriniaflavanone A (1) exhibited moderate cytotoxic activity.

[Synthesis and antitumor activity of fluoroquinolon-3-yl-s-triazole sulfide ketones and their derivatives from ciprofloxacin].

To discover an efficient strategy for the conversion of the antibacterial activity of fluoroquinolones into the antitumor activity, the three series of C-3 s-triazole-based derivatives including sulfide ketones (6a-6g), thiosemicarbazones (7a-7g) and fused heterocyclic thiazolotriazoles (8a-8g) were synthesized from ciprofloxacin (1), respectively. The structures were characterized by elemental analysis and spectral data. The antitumor activity was tested against three tumor cell lines (Hep-3B, Capan-1 and HL60) using the MTT assay. The three types of compounds all exhibited stronger anti-proliferative activities than ciprofloxacin in the test. The order of their activities was in compounds 7>8>6, and the order of selectivity against cancer cell lines was Capan-1, Hep-3B and HL60. Meanwhile, the SAR revealed that some compounds with electron-drawing group substituted such as fluoro- and nitro-phenyl compounds (6f, 7f, 8f) and (6g, 7g, 8g) displayed more significant activity than the control compounds, especially the IC50 values of thiosemicarbazone compounds 7f and 7g against Capan-1 was comparable to doxorubicin. Thus, a five-membered triazole as the C-3 bioisostere modified with the functionalized side-chain of sulfide-ketone thiosemicarbazone warrants special attention and further investigation.

[Inhibitory Effect of the Excretory/Scretory Proteins of Trichinella spiralis on Proliferation of Human Hepatocellular Carcinoma HepG2 Cell line].

Human hepatocellular carcinoma HepG2 Cell line were cultured with different concentrations of excretory/secretory proteins from Trichinella spiralis, and MTT assay was used to evaluate the cell inhibition rate. After co-cultured with 300 µg/ml excretory/secretory proteins for 24 h, the HepG2 cells were observed under a fluorescence microscope with AO and EB staining. When co-cultured with 75 µg/ml excretory/secretory proteins for 24 h, the HepG2 cells were quantified by flow cytometry using Annexin V-FITC/PI stain, and the expression of cleaved-caspase 9 was detected by immunofluorescence assay. The proliferation of HepG2 cells was inhibited significantly by excretory/secretory proteins in a dosage dependant manner. Under fluorescence microscope, some HepG2 cells presented typical apoptotic morphologic changes and the cleaved-caspase 9 protein expression was higher than that of the control. The early and late apoptotic cells and necrotic ones occupied 17.9%, 7.3%, and 6.6%, respectively.

A new species of the subgenus Stigmatochirus of Plastus Bernhauer (Coleoptera: Staphylinidae: Osoriinae) from Yunnan, China.

Plastus (Stigmatochirus) menglaius sp. nov. is described from Yunnan, China. Color images of the habitus and aedeagu of the new species are included. A key to the subgenus Stigmatochirus of World species is provided.

Pheromones emitted by both female and male moths regulate coordination between the sexes for Agriphila aeneociliella (Lepidoptera: Crambidae).

The complex and efficient sex pheromone communication system in insects is essential for reproduction and for reproductive isolation of species. In moths, sex pheromone communication starts with male attraction to compounds emitted by females; only a few species act in the reverse. However, how the pheromones that are emitted by both sexes co-regulate and coordinate mate finding and mating remains unknown. Here, we identified both the male and female pheromones of Eastern Grass Veneer moth, Agriphila aeneociliella (Lepidoptera: Crambidae), and demonstrated their efficiency in manipulating behavioral responses of the opposite sex. Combining data from analysis of gas chromatography-electroantennogram detection, gas chromatography-mass spectrometry, and olfactory behavior assays, the female pheromone of A. aeneociliella was identified as (Z,Z,Z)-9,12,15-octadecatrienal and (Z)-9-hexadecenyl acetate, while the male pheromone was determined to be 1-nonanal. Both the 2 individual components of the female pheromone and their binary mixture were significantly attractive to males, and the 1-nonanal male pheromone induced strong electrophysiological responses in females and induced attraction of females in a Y-tube olfactory test. Depending on the concentration of 1-nonanal, its addition to the binary mixture of the female pheromone either enhanced (10-3 or 10-2  µg/µL) or reduced (1 µg/µL) the aphrodisiac effect of the mixture on males. In wind-tunnel bioassays, different concentrations of pheromones, including the binary mixture of female pheromone and the mixture of male and female pheromones, had significant effects on male behavior. Our findings suggested that the blend of both female and male pheromones plays a significant role in the sexual communication system in some moths.

Fluorinated poly(p-triphenyl piperidine) anion exchange membranes with robust dimensional stability for fuel cells.

Anion exchange membrane fuel cells (AEMFCs), which are more economical than proton exchange membrane fuel cells (PEMFCs), stand out in the context of the rapid development of renewable energy. Superacid-catalyzed ether-free aromatic polymers have recently received a lot of attention due to their exceptional performance, but their development has been hampered by the trade-off between the dimensional stability and ionic conductivity of anion exchange membranes (AEMs). Here, we introduced fluoroketones containing different numbers of fluorinated groups (x = 0, 3 and 6) in the main chain of p-terphenyl piperidine because of the favorable hydrophobic properties of fluorinated groups. The results show that fluorinated AEMs can enhance OH- conductivity by building more aggregated hydrophilic channels while ensuring dimensional stability. The PTF6-QAPTP AEM with more fluorinated groups has the most excellent performance at 80 °C with an OH- conductivity of 142.7 mS cm-1 and a swelling ratio (SR) of only 4.55 %. Additionally, it exhibits good alkali durability, with the OH- conductivity and quaternary ammonium (QA) cation retaining at 93.45% and 92.6%, respectively, after immersion in a 2 M NaOH solution at 80 °C for 1200 h. In addition, the power density of the PTF6-QAPTP based single cell reaches 849 mW cm-2 when the current density is 1600 mA cm-2. The PTF6-QAPTP based cell has a voltage retention of 88% after 80 h of stability testing at a constant current density of 300 mA cm-2 at 80 °C.

A novel Vestitain A from the ripe fruits of Embelia vestita Roxb.

A new compound, vestitain A (1), together with 11 known compounds were isolated from the ripe fruits of Embelia vestita Roxb., among them compounds 5,10-12 were isolated from this plant for the first time. Their structures were elucidated and characterized by detailed spectroscopic analysis. Further, the isolated new compound 1 was evaluated for its hypoglycemic effects in vivo. Our research showed that compound 1 could decrease the fasting blood glucose (FBG) by approximately 36.31% in diabetic rats at the high dose (800 mg/kg). By the Morris Water Maze experiments, we found that compound 1 had the effect of intervention on social behavior in diabetic rats, which might provide a reference basis for its development and utilization as a potential hypoglycemic drug.

Nirmatrelvir/ritonavir use in patients with COVID-19 on hemodialysis: a case series.

Patients undergoing hemodialysis (HD) are particularly vulnerable to coronavirus disease 2019 (COVID-19) and are at increased risk of developing severe infection. However, given the exclusion of such patients from clinical trials, there are limited data regarding the effectiveness of the antiviral drug nirmatrelvir/ritonavir (N/R) in patients on HD. We prescribed N/R to 4 patients on HD with COVID-19 after obtaining informed consent. Their clinical symptoms were improved at approximately 3 days after N/R administration. The viral load was reduced after approximately 10 days. The main adverse effects were nausea and vomiting. Rational dosage adjustment obtained good tolerance but did not influence the efficacy. These results suggest that N/R may be a promising agent for patients on HD with COVID-19.

Effects of the crown ether cavity on the performance of anion exchange membranes.

Anion exchange membrane fuel cells (AEMFCs) have emerged as a promising alternative to proton exchange membrane fuel cells (PEMFCs) due to their adaptability to low-cost stack components and non-noble-metals catalysts. However, the poor alkaline resistance and low OH- conductivity of anion exchange membranes (AEMs) have impeded the large-scale implementation of AEMFCs. Herein, the preparation of a new type of AEMs with crown ether macrocycles in their main chains via a one-pot superacid catalyzed reaction was reported. The study aimed to examine the influence of crown ether cavity size on the phase separation structure, ionic conductivity and alkali resistance of anion exchange membranes. Attributed to the self-assembly of crown ethers, the poly (crown ether) (PCE) AEMs with dibenzo-18-crown-6-ether (QAPCE-18-6) exhibit an obvious phase separated structure and a maximum OH- conductivity of 122.5 mS cm-1 at 80 °C (ionic exchange capacity is 1.51 meq g-1). QAPCE-18-6 shows a good alkali resistance with the OH- conductivity retention of 94.5% albeit being treated in a harsh alkali condition. Moreover, the hydrogen/oxygen single cell equipped with QAPCE-18-6 can achieve a peak power density (PPD) of 574 mW cm-2 at a current density of 1.39 A cm-2.

[The sexual behavior characteristics and STD infection status of women who have sex with women in Beijing].

OBJECTIVE: To study sexual behavioral characteristics, infection status of HIV and sexually transmitted infections (STDs) among women who have sex with women (WSW) in Beijing. METHODS: A total of 150 WSWs were recruited by snowball sampling in several main types of activity sites of WSW (including NGOs, salons, bars, etc.) in Beijing during September 2010 to April 2011. Information on demographic factors, sexual behavioral characteristics, infection status of STDs, and use of medical care were collected by questionnaire investigation. Serum samples, vaginal and cervical swabs were taken for each subject to test HIV, hepatitis B (HBV), hepatitis C (HCV), syphilis, herpes simplex virus (HSV), neisseria gonorrhoeae, chlamydia trachoma, bacterial vaginosis, trichomonas vaginalis and mycotic vaginitis. RESULTS: The age range of the subjects was from 19 to 46 years old. Approximately 82.67% (124/150) of them aged 20 to 29 years old, and 54.00% (81/150) were unmarried. Approximately 76.67% (115/150) had an education level of college degree or above, and 70.67% (106/150) were migrants. Approximately 66.66% (100/150) of the subjects considered themselves as homosexual, 28.00% (42/150) as bisexual. The age at first homosexual sex ranged from 11 to 30 years old. The median of number of female sex partners in last 1 year was 1, and 33.78% (50/148) of the subjects had 2 or more female sex partners. Approximately 20.27% (30/148) of the subjects applied sex instruments, of which 66.67% (20/30) never or seldom used condoms. Hand-clitoris and hand-vagina contacts were the main types of woman-to-woman sex, accounting for 91.22% (135/148). 12.00% (18/150) of the subjects had sex with men during last year. Among them, 50.00% (9/18) used condoms at last heterosexual sex, and 66.67% (12/18) never or seldom used condoms at last heterosexual sex in past 1 year. One subject provided paid sex service for men, and used condoms every time. In the past one year, 45.33% (68/150) of the subjects had symptoms of STDs, but only 36.76% (25/68) of them sought medical care. The infection rate of STDs was 34.67% (52/150). The infection rates of NG and GV were both 16.11% (24/149), and those of mycotic vaginitis, chlamydia trachoma, and TV were 8.72% (13/149), 4.03% (6/149), and 0.67% (1/150), respectively. The numbers of subjects tested positive for HBV, HCV, and syphilis were all 1. No subjects were found HIV positive or HSV-2 positive. CONCLUSION: Multiple sexual partners and heterosexual sex behavior were common among WSW in Beijing. More than 1/3 of the subjects were infected with STD.

[Characteristics of sexual behaviors and infection status of AIDS and other sexually transmitted diseases among men who have sex with men in 2009 in Beijing].

OBJECTIVE: To analyze sexual behavior characteristics and sexually transmitted diseases such as AIDS infections in man who have sex with men (MSM) of Beijing Chaoyang district. METHODS: Through selecting 10 MSM as seed in Chaoyang district of the existing education member companion in its main activity place, using snowball sampling to select 1008 people that 16 years old or above, nearly a year had sex with male in Chaoyang to conduct an investigation.Questionnaires were used to investigate the general conditions, characteristics of sexual behaviors, condom usage, AIDS knowledge awareness and sexually transmitted diseases history. Each participant was asked to exsanguinated 5 ml peripheral blood for applying ELISA for HIV, syphilis helicoids detection and HIV positive samples confirm test. Using χ(2) test on the statistical analysis of the data. RESULTS: In the 1008 MSM, most of them were with the age of 20 - 29 years old (66.6%, 671/1008), giving priority to unmarried (79.1%, 797/1008), college degree and above culture during accounted for 43.3% (436/1008), 56.2% (566/1008) had both male and female sexual partners; the rate of temporary sexual partners who were one-night stand objects or friends or guest was 72.2% (226/313). The forever condom utilization rate was 29.0% (170/587) in MSM who had both male and female sexual partners, lower than that who had both male and female sexual partners (47.2%, 345/731). That the study objects of HIV infection who can use condom correctly rate was 3.6% (16/442), and that who never or occasionally used condom rate was 8.5% (24/228). AIDS, gonorrhea and other sexually transmitted diseases (to give priority to the acuteness wet wart) infection rates were 4.8% (6/124), 10.5% (13/124) and 8.9% (11/124) in those who had the history of group sex, and that was 5.9% (52/884), 1.9% (17/884) and 2.3% (20/884) in those who without the history of group sex. The research object's gonorrhea and other sexually transmitted diseases (to give priority to the acuteness wet wart) who provided commercial sex service infection rates were 7.7% (9/117) and 6.8% (8/117), significantly higher than those who did not provide a commercial sex service object of study subject (2.4% (21/891)and 2.6% (23/891)). CONCLUSION: HIV infection is high among MSM population. Types and identification of sexual partners, and condom use are associated with HIV and STD infection.

[Incidence and risk factors of HIV and syphilis seroconversion among men who have sex with men in Beijing].

OBJECTIVE: To study the incidence and risk factors of HIV and syphilis seroconversion among men who have sex with men (MSM) in Beijing. METHODS: A total of 550 MSM were recruited on the basis of community and followed up after 6 and 12 months in Beijing. Each subject was investigated by only one investigator at one time to collect information on demographics and behaviors. Blood samples were collected to test HIV and syphilis seroconversion. ELISA was used for screening test, west blotting (WB) and Particle agglutination were used for confirmatory test. RESULTS: A total of 550 MSM investigated, among which 4.5% (25/550) were HIV-positive and 29.3% (161/550) were syphilis-positive. For 525 HIV-negative MSM, 87.0% (457/525) retained during the 12-month investigation. Seroincidence for HIV and syphilis were 3.37/100 person-years (95%CI = 1.66 - 5.08) and 9.32/100 person-years (95%CI = 5.87 - 12.77) respectively. HIV seroconversions for those who performed and did not perform rectal douching after homosexual anal intercourse in the past 3 months were 7.11/100 and 0.76/100 person-years respectively. Multivariate Cox regression analysis revealed that rectal douching after homosexual anal intercourse in the past 3 months (HR = 9.23, 95%CI = 2.08 - 40.88) was significantly associated with HIV seroconversion. Syphilis seroconversions for those who met male sex partners in parks, public washrooms or bathhouses in the past 3 months were 41.77/100 and 7.97/100 person-years respectively. Syphilis seroconversions for those who performed and did not perform rectal douching after homosexual anal intercourse in the past 3 months were 16.17/100 and 4.92/100 person-years respectively. In the past 3 months, meeting male sex partners in parks, public washrooms or bathhouses (HR = 4.67, 95%CI = 1.77 - 12.34) and performing rectal douching after homosexual anal intercourse (HR = 3.09, 95%CI = 1.40 - 6.83) were significantly associated with syphilis seroconversion. CONCLUSION: The seroconversions of HIV and syphilis during the follow-up visits in this MSM cohort study in Beijing were very serious, and that the associated factors for seroconversions were rectal douching after homosexual anal intercourse and meeting male sex partners in parks, public washrooms or bathhouses.

[Regulatory effect of GDNF on the proliferation and differentiation of mammalian spermatogonial stem cells].

The glial cell-derived neurotrophic factor (GDNF) is a member of the transforming growth factor beta (Tgf-beta) superfamily, which is produced by Sertoli cells and plays an important role in the proliferation and differentiation of spermatogonial stem cells (SSC). The addition of proper amount of GDNF to the culture media can promote SSC proliferation in vitro. Besides, GDNF regulates the self-renewal and differentiation of SSCs through various signaling pathways. This review focuses on the effects of GDNF on the proliferation and differentiation of mammalian SSCs and GDNF-mediated signaling pathways.

Prevention and control of HIV/AIDS in China: lessons from the past three decades.

ABSTRACT: In the past 37 years, human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) has undergone various major transmission routes in China, with the world most complex co-circulating HIV-1 subtypes, even the prevalence is still low. In response to the first epidemic outbreak of HIV in injecting drug users and the second one by illegal commercial blood collection, China issued the Anti-Drug Law and launched the Blood Donation Act and nationwide nucleic acid testing, which has avoided 98,232 to 211,200 estimated infections and almost ended the blood product-related infection. China has been providing free antiretroviral therapy (ART) since 2003, which covered >80% of the identified patients and achieved a viral suppression rate of 91%. To bend the curve of increasing the disease burden of HIV and finally end the epidemic, China should consider constraining HIV spread through sexual transmission, narrowing the gaps in identifying HIV cases, and the long-term effectiveness and safety of ART in the future.