ICG-labeled PD-L1-antagonistic affibody dimer for tumor imaging and enhancement of tumor photothermal-immunotherapy.

In clinical practice, tumor-targeting diagnosis and immunotherapy against programmed death ligand 1 (PD-L1) have a significant impact. In this research, a PD-L1-antagonistic affibody dimer (ZPD-L1) was successfully prepared through Escherichia coli expression system, and conjugated with the photosensitizer of ICG via N-hydroxysuccinimide (NHS) ester to develop a novel tumor-targeting agent (ICG-ZPD-L1) for both tumor imaging diagnosis and photothermal-immunotherapy simultaneously. In vitro, ZPD-L1 could specifically bind to PD-L1-positive LLC and MC38 tumor cells, and ICG-ZPD-L1-mediated photothermal therapy (PTT) also showed excellent phototoxicity to these tumor cells. In vivo, ICG-ZPD-L1 selectively enriched into the PD-L1-positive MC38 tumor tissues, and the high-contrast optical imaging of tumors was obtained. ICG-ZPD-L1-mediated PTT exhibited a potent anti-tumor effect in vivo due to its remarkable photothermal properties. Furthermore, ICG-ZPD-L1-mediated PTT significantly induced the immunogenic cell death (ICD) of primary tumors, promoted maturation of dendritic cells (DCs), up-regulated anti-tumor immune response, enhanced immunotherapy, and superiorly inhibited the growth of metastatic tumors. In addition, ICG-ZPD-L1 showed favorable biosafety throughout the brief duration of treatment. In summary, these results suggest that ICG-ZPD-L1 is a multifunctional tumor-targeting drug integrating tumor imaging diagnosis and photothermal-immunotherapy, and has great guiding significance for the diagnosis and treatment of clinical PD-L1-positive tumor patients.

Successful treatment by a chlorin e6 derivative mediated photodynamic therapy combined holmium laser for cervical and vaginal giant condyloma acuminata and low-grade intraepithelial neoplasia:A case report.

Photodynamic therapy (PDT) is proved effective for treating low-grade squamous intraepithelial lesions (LSIL) and condylomata acuminata (CA). 5-Aminolevulinicacid (5-ALA) is the most common applied photosensitizer, but high rate of unbearable pain and relative long incubation time were reported. Here, we report a 27-year-old woman suffering from cervical and vaginal giant CA with LSIL involving the whole right vaginal fornix, cervical surface, and vaginal wall. Holmium yttrium aluminum garnet (Ho: YAG) laser was first applied to remove the giant CA lesions. STBF, a derivative of chlorin e6 (Ce6) was then applied on suspicious lesions as a new photosensitizer for 1 h. Lesions were exposed to LED illumination with a wavelength of 630 nm and light dose of 200-284 J/cm2 for cervical canal and the vaginal surfaces, 100-150 J/cm2 for cervix surface. Vaginal giant CA and LSIL lesions got complete remission at 6-month follow-up. Mild tolerable adverse reactions were observed after STBF-PDT and relieved in 24 h. Thus, the combination of Ho: YAG laser and STBF-PDT may be a novel option for cervical and vaginal giant CA and LSIL, especially for special vaginal fornix areas.

The combination of holmium laser and ALA-PDT for Bowenoid Papulosis with diffuse large B-cell lymphoma.

Bowenoid Papulosis (BP) is an anogenital pre-malignancy. BP with immunosuppression may recur, worsen, or possibly evolve into squamous cell carcinoma or Bowen's disease (BD), and it may also become resistant to conventional treatment. Here, we describe a complex case of BP together with BD and Diffuse Large B-Cell Lymphoma that was effectively treated with a holmium laser in conjunction with 5-Aminolevulinic Acid Photodynamic Therapy (ALA-PDT). The lesion totally vanished and the affected area remained intact with no recurrence at five years.

Modified painless photodynamic therapy for facial multiple actinic keratosis in China: A prospective split-face control study.

BACKGROUND: Aminolevulinic acid photodynamic therapy (ALA-PDT) is an effective treatment for multiple actinic keratosis (AK). However, PDT-induced pain often discontinues the therapy to reduce its efficacy, limiting its application. If modified painless PDT schedule with shorter photosensitizer dressing and higher dose illumination could achieve good efficacy in AK, it is still unknown. OBJECTIVES: To explore the efficacy and pain tolerance of the modified painless PDT (M-PDT) in facial multiple AK. METHODS: A split-face controlled clinical study including 14 patients with facial multiple AK was conducted. The patients received conventional PDT (C-PDT) on the left and M-PDT in the contralateral area. The left area (C-PDT) was illuminated by a red light-emitting diode light (144 J/cm2 ) after applying the 10% ALA cream for 3 h; the other had illumination for a total light dose of 288 J/cm2 after applying the 10% ALA cream for 0.5 h. The primary endpoint was the lesion clearance rate at 1-month postthree sessions of PDT. Secondary endpoints included pain scores, the incidence of adverse events during treatment, and cosmetic outcomes. RESULTS: At 1 month following three treatments, the total lesion clearance rate was comparable between M-PDT and C-PDT (91.6% vs. 89.0%). While the lesion clearance rate of M-PDT was higher than that of C-PDT in the Grade III lesions (86.5% vs. 72.0%, respectively) (p < 0.05). M-PDT achieved a 100% lesion clearance rate for Grade I lesions earlier than C-PDT, with M-PDT treated twice and C-PDT treated thrice. Moreover, the pain score during illumination was significantly lower for M-PDT than for C-PDT (p < 0.01). Regarding photoaging, the Global Subjective Skin Aging Assessment score showed that the total and atrophy scores of C-PDT and M-PDT were significantly improved, and M-PDT also reduced discoloration. There was no significant difference in adverse reactions between C-PDT and M-PDT. CONCLUSIONS: M-PDT is comparable to C-PDT's efficacy for treating facial multiple AK, resulting in much lower pain scores.

Successful treatment of non-melanoma skin cancer in three patients with Xeroderma Pigmentosum by modified ALA-PDT.

Xeroderma pigmentosum(XP) is a rare autosomal recessive genodermatosis. Individuals with XP are characterized by severe skin sensitivity to sunlight, and more susceptible to the development of skin malignancies in sun-exposed regions. We report the experience of modified 5-aminolaevulinic acid photodynamic therapy (M-PDT) in the treatment of three children with XP. They all developed multiple freckle-like hyperpigmented papules and plaques on the face from an early age. Multiple cutaneous squamous cell carcinoma (cSCC) and actinic keratosis (AK) were developed in case 1 and case 2, and basal cell carcinoma (BCC) was observed in case 3. Sanger sequencing of targeted gene identified that case 1 and case 3 carried compound heterozygous mutations, and case 2 carried a homozygous mutation in the XPC gene. After multiple courses of M-PDT, the lesions were removed with mild adverse reactions, nearly painless and satisfactory safety.

Local narrow margin excision sequential with modified ALA-PDT for successful treatment of an 86-year-old patient with malignant proliferating trichilemmal tumor.

Malignant proliferating trichilemmal tumor (MPTT) is thought to represent the malignant counterpart of benign proliferative pilar cyst, a lesion originating from the outer hair root sheath. We report a case of an 86-year-old woman with an exophytic nodule containing an ulcerated surface in the temporal region. Histopathologically confirmed MPTT without metastasis, and considering the risk of metastasis and recurrence, we used single local narrow margin excision combined with modified 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT). No recurrence of skin lesions was found in the 2-year follow-up after combined therapy. In conclusion, local narrow margin excision sequential with modified ALA-PDT may be a particularly promising and effective treatment option for MPTT.

Disubstituted pyrimidine-5-carboxamide derivatives as novel HIV-1 NNRTIs: Crystallographic overlay-based molecular design, synthesis, and biological evaluation.

Herein, via crystallographic overlay-based molecular hybridization strategy, a series of disubstituted pyrimidine-5-carboxamide derivatives were designed by introducing an amide moiety to the central core of the lead etravirine. All the newly synthesized compounds were evaluated for their anti-HIV-1 potencies in MT-4 cells using the MTT method. Most of the synthesized compounds displayed promising antiviral activities against the wild-type (IIIB) and a panel of HIV-1 NNRTIs-resistant strains. Especially, 21c exhibited the most potent activity (EC50 = 0.009-0.065 µM) against HIV-1 IIIB, L100I, K103N, Y181C, Y188L, and RES056, being comparable to those of etravirine. The inhibitory activity to reverse transcriptase (RT) was evaluated by ELISA method, and the target of the compounds was proved to be RT. Moreover, the molecular docking was investigated to clarify the binding mode of 21c with RT. Overall, the results demonstrated that 21c could serve as a lead for further modification to develop novel HIV-1 NNRTIs.

Identification of Novel Diarylpyrimidines as Potent HIV-1 Non-Nucleoside Reverse Transcriptase Inhibitors by Exploring the Primer Grip Region.

HIV-1 reverse transcriptase (RT) plays a crucial role in the viral replication cycle, and RT inhibitors can represent a promising pathway in treating AIDS. To explore the primer grip region of HIV-1 RT, using -CH2O- as a linker, substituted benzene or pyridine rings were introduced into the left wing of diarylpyrimidines (DAPYs). A total of 17 compounds with new structures were synthesized. It showed that all compounds exhibited anti-HIV-1 (wild-type) activity values ranging from 7.6−199.0 nM. Among them, TF2 (EC50 = 7.6 nM) showed the most potent activity, which was better than that of NVP (EC50 = 122.6 nM). Notably, compared with RPV (CC50 = 3.98 µM), TF2 (CC50 > 279,329.6 nM) showed low cytotoxicity. For HIV-1 mutant strains K103N and E138K, most compounds showed effective activities. Especially for K103N, TF2 (EC50 = 28.1 nM), TF12 (EC50 = 34.7 nM) and TF13 (EC50 = 28.0 nM) exhibited outstanding activity, being superior to that of NVP (EC50 = 7495.1 nM) and EFV (EC50 = 95.1 nM). Additionally, TF2 also showed the most potent activity against E138K (EC50 = 44.0 nM) and Y181C mutant strains (EC50 = 139.3 nM). In addition, all the compounds showed strong enzyme inhibition (IC50 = 0.036−0.483 µM), which demonstrated that their target was HIV-1 RT. Moreover, molecular dynamics simulation studies were implemented to predict the binding mode of TF2 in the binding pocket of wild-type and K103N HIV-1 RT.

Identification of Boronate-Containing Diarylpyrimidine Derivatives as Novel HIV-1 Non-Nucleoside Reverse Transcriptase Inhibitors.

In this study, privileged boronic acid ester was introduced into the right wing of etravirine (ETR) to obtain a series of novel boronate-containing derivatives. These newly synthesized derivatives were evaluated for their anti-HIV potency in MT-4 cells using the MTT method, and their inhibitory activity to HIV-1 reverse transcriptase (RT) was assayed by the ELISA method. Most of the synthesized compounds displayed promising antiviral activity against the wild-type and a wide range of HIV-1 mutant strains. In particular, 4a exhibited the most potent activity against the wild-type and a panel of single mutations (L100I, K103N, Y181C, and E138K) with EC50 values ranging from 0.005 to 0.648 µM, which were much superior to those of nevirapine (EC50 = 0.151 µM). Moreover, 4b turned out to be an effective inhibitor against the double-mutant strains F227L + V106A and RES056 with EC50 values of 3.21 and 2.30 µM, respectively. RT inhibition activity and molecular docking were also investigated.

Vulvar lichen sclerosus progressing to squamous cell carcinoma due to the poor compliance for the follow-up after ALA-PDT.

5-Aminolevulinic acid photodynamic therapy (ALA-PDT) is an effective modality for vulvar lichen sclerosus (VLS) and squamous cell carcinoma (SCC) in situ. Here we first describe a case of VLS that relapsed after ALA-PDT without follow-up and progressed to SCC eventually. The first two courses of ALA-PDT (total of 21 sessions) were effective for VLS. The third course of ALA-PDT (2 sessions) was changed to the surgery because of SCC was diagnosised. In our case, we emphasis that maintain regular follow-up to treat the recurrent lesions to prevent the development of SCC.

Modified photodynamic therapy of vulvar giant condyloma acuminatum complicated by systemic lupus erythematosus: A case report.

Giant condyloma acuminatum (GCA) is a rare disease characterized by cauliflower-like tumors in the genital area, with a peculiar smell, bleeding, and local infection. Its occurrence is related to a variety of risk factors, such as human papillomavirus infection, immune deficiency, poor sanitary conditions, multiple sexual partners, and chronic genital infection. Surgical resection is still the preferred treatment for the disease. This paper reports that a patient with GCA complicated by systemic lupus erythematosus (SLE) and persistent thrombocytopenia who could not be treated surgically was treated with modified photodynamic therapy (M-PDT) in our outpatient department. After several treatments, the pain, odor, and secretion on the surface of the wart were significantly reduced, although the size of the wart was not significantly improved. Unfortunately, the patient died subsequently due to the aggravation of SLE. However, clinical treatment with M-PDT may be used as a palliative treatment when similar patients are encountered.

Identification of key genes and infiltrating immune cells among acetaminophen-induced acute liver failure and HBV-associated acute liver failure.

Background: Acute liver failure (ALF) is a life-threatening complication that is relatively uncommon. ALF causes severe hepatocyte damage and necrosis, which can lead to liver dysfunction and even multi-organ failure. A growing body of evidence suggests that immune cell infiltration and some abnormally expressed genes are associated with ALF development. However, in ALF, they have yet to be thoroughly investigated. Methods: The Gene Expression Omnibus (GEO) database was used to obtain microarray datasets such as GSE74000, GSE120652, GSE38941, and GSE14668, which were then examined via GEO2R to determine differentially expressed genes (DEGs) associated with ALF. Metascape was employed to annotate the underlined genes using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. The mechanism of IGF1 in 2 different kinds of ALF including acetaminophen-induced ALF and hepatitis B virus (HBV)-induced ALF was studied using gene set enrichment analysis (GSEA). Next, immune cell infiltration was investigated and differentiated in ALF using CIBERSORT. Results: Six genes (HAO2, IGF1, PLA2G7, SC5D, GNE, SLC1A1) were found to be abnormally expressed in the 2 distinct types of ALF i.e., acetaminophen-induced ALF and HBV-induced ALF. IGF1 was identified as a hub gene in ALF and was found to be associated with several developmental cascades including immune responses, inflammatory responses, and intracellular calcium homeostasis. Additionally, the number of CD4 naive T cells, CD8 T cells, and follicular helper T cells was increased in acetaminophen-induced ALF, whereas the number of activated NK cells, resting NK cells, and plasma cells was increased in HBV-induced ALF. Conclusions: The present study determined a potential molecular target, namely IGF1, in acetaminophen-induced ALF and HBV-induced ALF, which may provide novel insights into the pathophysiology and management of ALF. Concurrently, the putative immunological pathways have been found.

ERß overexpression may not be a direct prognostic factor in patients with NSCLC: A meta-analysis.

Overall survival of non-small cell lung cancer (NSCLC) patients remains disappointingly low. The estrogen receptor (ER) was considered a promising therapeutic target for NSCLC. Numerous studies have linked expression of ERß to lung cancer outcome. However, results are conflicting regarding the association of ERß with surviving lung cancer. The aim of this meta-analysis was to evaluate the prognostic aspect of ERß expression on survival among NSCLC patients. We performed a final analysis of prognostic value of overexpression ERß on 3500 patients from 18 evaluable studies (from January 1, 2000 to May 1, 2021). The reference category is specified as low ERß expression levels. Summarized hazard ratios were calculated. Our study showed that the pooled hazard ratios of ERß overexpression for overall survival in NSCLC was 0.81 (95% confidence interval (CI): 0.64-1.02, P = 0.07) by univariate analysis and 1.06 (95% CI: 0.83-1.36, P = 0.63) by multivariate analysis. Pooled hazard ratio by univariate analysis in Asian studies was 0.73 (95%CI: 0.59-0.89, P = 0.002). Pooled hazard ratio by univariate analysis was 0.75 (95% CI: 0.61-0.93, P = 0.009) from seven studies reported for nuclear ERß. No significant results were found in subgroups by multivariate analysis. No significant results were found in studies outside Asia or in studies reported for cytoplasmic ERß. Our results suggested that expression of ERß might not be a direct prognostic factor for NSCLC patients. More detailed prospective studies are needed to identify direct prognostic factors in these patients.

Acute acne flare following isotretinoin administration successfully treated by 5-aminolevulinic acid photodynamic therapy.

The acne flare occurs in a small proportion of patients at the initiation of oral isotretinoin. The traditional therapy is oral corticosteroids. But for particular patients it also can be accompanied by side effects, such as metabolic disorder, inhibition of immune function among other effects. Here, we report a young man with acute acne flare following isotretinoin administration treated by 5-aminolevulinic acid photodynamic therapy (ALA-PDT). 5-ALA cream (5%) was applied on all skin lesions and kept under a black plastic film for 2 h, and then the skin lesions were exposed toLED PDT illumination with a wavelength of 633 ± 6 mm, power density of 42 mW/cm2, irradiation duration of 30 min for each light spot and light dose of 75.6 J/cm2, for each treated area. A total of seven treatments were provided, with complete clearance and excellent cosmetic result. Here, ALA-PDT as a novel therapeutic option in acne flare.

Fluorescence kinetics study of twice laser irradiation based HpD-PDT for nonmelanoma skin cancer.

BACKGROUND: Hematoporphyrine injection (HpD)-based photodynamic therapy (HpD-PDT) has emerged as a promising cancer therapy. However, its tumor-targeting ability and metabolokinetics in nonmelanoma skin cancer (NMSC) have not been well explored. Importantly, photodynamic diagnosis is widely used for cancer lesion assessment and positioning to ensure effective therapy, while the photosensitizer metabolic kinetics study is utilized for biosafety assessment and light-protection instruction. These are particularly important for the optimization of therapeutic parameters. OBJECTIVES: In the present study, NMSC patients were subjected to twice laser irradiation-based HpD-PDT strategy. Broadly, the study aimed to assess long-term variations in fluorescence (FL) intensity in vivo in NMSC patients after intravenous (i.v.) administration of HpD, and thus obtain information regarding metabolism, biosafety, and light-protection instruction for HpD during the therapy. METHODS: In vitro experiments were used for the evaluation of absorption and fluorescent characterization of HpD in aqueous solution and cutaneous squamous cell carcinoma (SCC) cells. For in vivo assessment, 20 patients with NMSC, including SCC, basal cell carcinoma (BCC), Bowen disease (BD), extramammary Paget's disease (EMPD), and malignant proliferating tricholemmoma (APT), were recruited, and treated with HpD-PDT. To evaluate the selectivity and pharmacokinetics of HpD in vivo, relative changes in FL intensity for lesional, perilesional, and nonlesional skin of nonmelanoma skin cancer patients, before and after HpD injection, were semiquantitatively analyzed for 1 month, using the FL detection system and Wood's lamp. RESULTS: The absorption and FL spectra were detected and semiquantitatively analyzed in HpD diluted solution and SCC cells after coincubation with HpD. After i.v. administration of HpD in EMPD patients, FL was detected in the skin lesions at 24 hours, and it was characterized by clear edges. Importantly, FL intensity in the skin lesions increased significantly at 48 and 72 hours postinjection, which was suitable for HpD-PDT. After 72 h, it decreased gradually and reached close to the baseline value at 4 weeks postinjection. No severe side effects were observed during HpD injection and the therapy. Urinary tract infection was recorded in one patient (with a previous history of recurrent urinary tract infections) after HpD-PDT, and the patient was cured afterward. Transient light was observed in two patients after HpD-PDT and they soon recovered after therapy. CONCLUSIONS: The present study reported a significant increase in FL intensities at 48 and 72 hours after i.v. administration of HpD in patients with nonmelanoma skin cancers, which indicated accumulation of HpD at the cancer site. Importantly, HpD was found to be safe for NMSC patients. After therapy, FL intensities decreased, which indicated expending and metabolization of HpD. Thus, the results of the present study highlighted the suitability of a twice red-light laser irradiation strategy for the application of HpD-PDT in nonmelanoma skin cancer treatment.

Photodynamic therapy of intravenous injection combined with intratumoral administration of photosensitizer in squamous cell carcinoma.

Cutaneous squamous cell carcinomas (cSCC) is one of the most common types of non-melanoma skin cancer (NMSC), and surgical excision is the primary regimen in most cases. However, in some circumstances of special lesion locations like lips, eyelids or vulva, old age or patient choice non-surgical therapy may be alternative. This is a case of one 93-year-old female cSCC patient who declined surgery. Treatment of hematoporphyrin derivatives photodynamic therapy (HpD-PDT) consisting of both intravenous and topical photosensitizers plus red light irradiation was prescribed. Clinical remission was achieved without evidence of recurrence and most cosmetic function was preserved.

A prospective study of adverse reactions of ALA-PDT for acne vulgaris.

BACKGROUND: Acne vulgaris is a chronic inflammatory skin disease around pilosebaceous unit. 5-Aminolaevulinic acid photodynamic therapy (ALA-PDT) is an effective therapy for severe acne vulgaris. However, the lack of detailed information of adverse reactions limits the promotion of ALA-PDT in clinic. OBJECTIVE: To systemically investigate the adverse reactions relating to ALA-PDT for acne vulgaris. METHODS: A prospective study was performed at the Shanghai Skin Disease Hospital. RESULTS: In the prospective study, 35 patients with acne vulgaris completed the trial. The adverse reactions were first divided into acute-phase adverse reactions, including erythema (94.3%), post-treatment pain (91.4%), burning skin (91.4%), dry skin (91.4%), itching (85.7%), pustule (82.9%), edema (20%) and blister (11.4%), or recovery-phase adverse reactions, which included crust (65.6%), exudation (48.6%) and hyperpigmentation (42.7%). Younger patients were more likely to have pustules (P<0.05). Male patients were represented itching more often than female patients (P<0.05). The efficacy of ALA-PDT for severe acne was positively related to the severity of acute-phase adverse reactions. The duration of pain was shortest. Crusting and hyperpigmentation lasted considerably longer. CONCLUSION: In the present study, we recorded the relative incidence of various adverse reactions following ALA-PDT for acne vulgaris. The severity of adverse reactions tended to decrease with increased patient age, except for itching and hyperpigmentation. Light-to-moderate adverse reactions might be the inflammatory reactions of ALA-PDT, predicting a good efficacy. A form for evaluation of adverse reactions based on the present study could assist dermatologists in predicting and managing adverse reactions for greater efficacy and higher patient satisfaction.

Adverse reactions of ALA-PDT for the treatment of cutaneous diseases: A retrospective study.

BACKGROUND: 5-Aminolaevulinic acid photodynamic therapy (ALA-PDT) is an effective therapy for cutaneous diseases, such as precancers, superficial non melanoma skin cancers and certain inflammatory or viral conditions. However, the absence of a complete picture of adverse reactions limits the promotion of ALA-PDT. OBJECTIVE: To systemically investigate the detailed evidence of adverse reactions relating to ALA-PDT for skin diseases. METHODS: A retrospective study performed at the Shanghai Skin Disease Hospital. RESULTS: In the retrospective study, 439 patients were included. Incidences of adverse reactions, including in-treatment pain (98.8%), erythema (92.4%), edema (35.0%), exudation (23.0%), hyperpigmentation (27.3%) were clarified. Edema was more common in female patients (P<0.05). Patients with HPV-related skin diseases were more likely to suffer erythema, edema or exudation (P<0.05). Hyperpigmentation was more likely to occur in skin appendage disorders (P<0.05). Fever (2.4%) and hypopigmentation (1.9%) are two neglected adverse reactions analyzed in detail. Fever is more prevalent in female patients. Hypopigmentation occurred predominantly in elderly with skin cancer or precancerosis lesions. CONCLUSION: The results outline detailed information about the adverse reactions, including systemic reactions following ALA-PDT, assisting dermatologists in predicting and managing adverse reactions for greater efficacy and higher patient satisfaction.

Efficacy and therapeutic reactions of tri-needle-pretreatment combined with painless ALA-PDT for the treatment of moderate-to-severe acne vulgaris: A randomized controlled trial.

BACKGROUND: Our clinical experience suggests that pretreatment of the original lesions may be crucial for enhancing the efficacy of 5-aminolevulinic acid-photodynamic therapy (ALA-PDT) in moderate-to-severe acne vulgaris. We performed this randomized controlled trial (RCT) to validate this observation. METHODS: Efficacy and therapeutic reactions between tri-needle-pretreatment with ALA-PDT (TP-PDT) and conventional ALA-PDT without pretreatment (NP-PDT) were compared. In TP-PDT group, tri-needle-pretreatment was performed using comedone extractors, fire needles, or plum-blossom needles, according to the lesion type. In the TP-PDT group, 5% ALA cream was applied to lesions 30 min before illumination (LED red light: 633±10 nm, 40 mW/cm2, 150 J/cm2). In the NP-PDT group, 5% ALA cream was applied 1 h before illumination (60 mW/cm2, 72 J/cm2). Patients underwent four sessions, at 1-week intervals. The efficacy was evaluated as the proportion of patients achieving a remarkable effective rate, based on the reduction in the number of lesions. A numeric rating scale was used to assess the severity of pain, erythema, and edema. RESULTS: Forty-eight patients completed the trial. The proportion of remarkable effective rate was significantly greater and the pain score was significantly lower for the TP-PDT than NP-PDT group. The edema score was significantly higher for the TP-PDT than NP-PDT group. There was no difference in erythema scores between the two groups. CONCLUSION: The tri-needle-pretreatment can improve the efficacy of ALA-PDT, without an increase in pain, for the treatment of moderate-to-severe acne vulgaris. These qualities make the TP-PDT a promising gold standard pretreatment for ALA-PDT for acne vulgaris.