[Role of intercellular adhesion molecule-1 in adhesion and migration of mesenchymal stem cells in ankylosing spondylitis and its mechanisms].

Objective: To investigate the role and underlying mechanisms of intercellular adhesion molecule-1 (ICAM-1) in the adhesion and migration of mesenchymal stem cells (MSCs) in patients with ankylosing spondylitis (AS). Methods: Bone marrow and ligament tissues were collected during surgery from patients with AS and thoracolumbar fractures (as controls, HC) treated from October 2021 to October 2022 at Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital. MSCs were isolated and cultured from the bone marrow using the Ficoll separation method. Cell morphology was observed under high-resolution microscopy, and differences in the cytoskeletal features between AS-and HC-MSCs were analyzed through immunofluorescence staining. The expression of ICAM-1 was quantified in both groups using real-time quantitative polymerase chain reaction (RT-qPCR) and flow cytometry. Transwell migration assays and wound healing experiments were conducted to evaluate the differences in migration rates between the two groups of MSCs. Results: The interspinous ligament and bone marrow was acquired in AS (2 males and 1 female; 33, 37, 32 years old, respectively) and no-AS patients (2 males and 1 female; 35, 32, 38 years old, respectively). AS-MSCs exhibited broader cell morphology compared to HC-MSCs under bright field and fluorescence microscopy. Immunofluorescence staining of the interspinous ligament showed higher expression of ICAM-1 (68.38±3.42 vs 48.31±2.43) and CD105 (37.97±2.16 vs 23.36±2.06) in AS patients (both P<0.001). Western blot and RT-qPCR analysis revealed significantly stronger protein expression and transcription levels of ICAM-1 in AS-MSCs when compared to those in HC-MSCs (both P<0.001). Flow cytometry confirmed greater mean fluorescence intensity of ICAM-1 in AS-MSCs than in that in HC-MSCs (924.30±54.99 vs 636.47±40.03, P=0.002). Regarding cell adhesion efficiency, it showed no significant difference between AS-MSCs and HC-MSCs in the early stage of adhesion (0.5 h: 1 496±213 vs 1 205±163, P=0.133), but they were all significantly higher in AS-MSCs in the later stage (1 h: 2 894±172 vs 1 908±155, P=0.002; 2 h: 4 540±286 vs 3 334±188, P=0.004; 3 h: 5 212±281 vs 4 208±303, P=0.014). Finally, cell migration experiments demonstrated a stronger migration capability of AS-MSCs compared to HC-MSCs (5 449±172 vs 4 016±155, P<0.001), and the inhibition efficiency of A-205804 on the migration rate of AS-MSCs was stronger than that on HC-MSCs (2 145±239 vs 3 539±316, P=0.004). Conclusions: The aberrant expression of ICAM-1 markedly influences the adhesion and migration dynamics of MSCs. Elevated ICAM-1 levels in MSCs derives from patients with AS significantly enhance their migratory capabilities.

[Advances in the regulation of craniomandibular regeneration by topical application of parathyroid hormone].

Parathyroid hormone (PTH) is a polypeptide molecule synthesized and secreted by parathyroid principal cells. It is an important hormone to maintain the balance of calcium and phosphorus metabolism in the body. It has the dual function of promoting bone formation and bone resorption. In clinic, it promotes osteogenesis by intermittent low-dose subcutaneous injection. In order to avoid the problems of subcutaneous injection, such as poor patient compliance, low utilization of target organs and pain at the injection site, the local application of PTH has attracted much attention in recent years. However, how to realize the local application of PTH and the effect of the local application need to be confirmed by more experiments. This article reviews the local application of PTH and the promotion of jaw regeneration in recent years, in order to provide reference for the local application and research of PTH.

[Interleukin-9 promotes the activation of hepatic stellate cells in mice infected with Schistosoma japonicum].

OBJECTIVE: To investigate the effects of interleukin-9 (IL-9) in the activation of hepatic stellate cells (HSCs) in mice infected with Schistosoma japonicum. METHODS: Primary HSCs were isolated from mice 7 weeks post-infection with S. japonicum using the in situ liver perfusion and density gradient centrifugation, and cultured in vitro. HSCs were randomly assigned to the PBS control group and IL-9 stimulation group (stimulation with 20 ng/mL IL-9). HSCs were harvested 48 h and 72 h poststimulation, and the expression of α-smooth muscle actin (α-SMA), type I collagen (Col I) and type III collagen (Col III) was determined in HSCs using Western blotting. RESULTS: Following stimulation with 20 ng/mL IL-9 for 48 h, the expression of α-SMA [(0.87 ± 0.02) vs. (0.69 ± 0.01); t = 17.39, P < 0.01], Col I [(0.74 ± 0.02) vs. (0.65 ± 0.01); t = 9.56, P < 0.01] and Col III [(0.94 ±0.04) vs. (0.75 ± 0.03); t = 6.15, P < 0.01] was significantly greater in HSCs in the IL-9 stimulation group than in the PBS control group. Following stimulation with 20 ng/mL IL-9 for 72 h, the expression of α-SMA was significantly greater in HSCs in the IL-9 stimulation group than in the PBS control group[(0.76 ± 0.02) vs. (0.58 ± 0.02); t = 12.52, P < 0.01]; however, there were no significant differences between the two groups in terms of Col I [(0.68 ± 0.02) vs. (0.66 ± 0.02); t = 1.15, P > 0.05] or Col III expression [(0.75 ± 0.01) vs. (0.72 ± 0.02); t = 2.22, P > 0.05]. CONCLUSIONS: IL-9 promotes the activation of HSCs in mice infected with S. japonicum.

[Expression of osteoprotegerin and receptor activator of nuclear factor kappa-B ligand in mandibular ramus osteotomy healing with administration of different doses of parathyroid hormone].

Objective: To investigate the effect of parathyroid hormone (PTH) on the bone healing of mandibular ramus osteotomy. Methods: The mandibular ramus osteotomy model was established in sixty rabbits and these rabbits were randomly divided into experimental group A, experimental group B and control group. In the experimental group A and experimental group B, the rabbits were given PTH (20 and 40 µg/kg respectively) every other day after operation. In the control group, 1 ml saline was given. The animals were sacrificed at 1 week, 2 weeks, 3 weeks and 4 weeks postoperatively. The new bone formation was observed by histology and cone bone CT. The expression of osteoprotegerin and receptor activator of nuclear factor kappa-B (RANKL) in the new bone was detected by real-time quantitative PCR. Results: The experimental groups has better osteogenesis and the bone mineral density than the control group in osteotomy area. The experimental group B showed the best osteogenesis.Osteoprotegerin mRNA expression in experimental group A (1.127±0.035, 1.742±0.049, 1.049±0.062, 1.063±0.036) was significantly higher than that in the control group in each period (0.965±0.082, 1.254±0.071, 0.793±0.061, 0.684±0.055) (P=0.010, P=0.000, P=0.001, P=0.020), while group B (1.416±0.205, 2.648±0.168, 1.652±0.091, 1.712±0.070) was significantly higher than group A (P=0.000, P=0.010, P=0.023, P=0.003). RANKL mRNA expression in control group (1.666±0.086, 1.058±0.105, 0.885±0.124, 0.972±0.136) was significantly higher than that of the group A (0.788±0.036, 0.585±0.017, 0.692±0.017, 0.527±0.051) (P=0.001, P=0.006, P=0.003, P=0.028) in each period, while group A was significantly higher than group B(0.247±0.022, 0.240±0.034, 0.134±0.011, 0.103±0.050) (P=0.000, P=0.001, P=0.002, P=0.012). Conclusions: PTH can upregulate the expression of osteoprotegerin and reduce expression of RANKL, thus promoting new bone formation. Intermittent administration of high dose of parathyroid hormone can further promote the healing process after mandibular ramus osteotomy.

[The progress in application of parathyroid hormone in craniomaxillofacial bone regeneration study].

Parathyroid hormone(PTH)is synthesized and secreted by chief cell of Gley's glands which possesses dual functions of catabolism and anabolism. It regulates the proliferation and differentiation of multiple cell lines including osteoblast, osteoclast and skeletal lining cells. Furthermore, PTH activates various signaling pathways which control calcium, phosphorous' metabolism and bone conversion, accelerating the bone regeneration and reconstruction. However, the study of PTH in craniomaxillofacial bone regeneration is relatively less and whether the role of parathyroid glands and the mechanism of ossification are consistent with the long bone or not needs further investigation. This review focuses on the progress of PTH in craniomaxillofacial bone regeneration in recent years.

Facile preparation of free-standing rGO paper-based Ni-Mn LDH/graphene superlattice composites as a pseudocapacitive electrode.

A novel film electrode was assembled via a simple filtration process, with an rGO paper as the substrate and Ni-Mn LDH/graphene superlattice composites as the functional layer. The electrode presented typical pseudocapacitive behaviours with excellent rate property and cycle stability.

Recurrent splice-site mutation in MBTPS2 underlying IFAP syndrome with Olmsted syndrome-like features in a Chinese patient.

Mutations in MBTPS2 have been reported to cause a broad phenotypic spectrum of X-linked genodermatoses, including IFAP (ichthyosis follicularis; atrichia and photophobia) syndrome (OMIM 308205) with or without BRESHECK (brain anomalies, retardation of mentality and growth, ectodermal dysplasia, skeletal malformations, Hirschsprung disease, ear deformity and deafness, eye hypoplasia, cleft palate, cryptorchidism, and kidney dysplasia/hypoplasia) syndrome, keratosis follicularis spinulosa decalvans (KFSD; OMIM 308800) and an X-linked form of Olmsted syndrome. We report a recurrent intronic mutation in MBTPS2 (c.671-9T>G) in a Chinese patient with the typical triad of IFAP syndrome (i.e. ichthyosis, atrichia and photophobia), along with pachyonychia, palmoplantar and periorificial keratoderma, which were reminiscent of Olmsted syndrome. Interestingly, this mutation was previously reported in two cases of IFAP without keratoderma, which suggests clinical heterogeneicity of the same mutation in MBTPS2. The concomitance of Olmsted syndrome-like features in this patient with IFAP may challenge the existence of the X-linked form of Olmsted syndrome as an independent condition.

Sequence characterization and promoter identification of porcine APC10 gene.

APC10 protein, a subunit of the anaphase-promoting complex (APC), plays an essential role in the progression of cells from mitosis to G1. In this study, we cloned and sequenced partial cDNA, intron 1 and 5'-flanking sequences of porcine APC10. The partial cDNA is 595 bp long and has an open reading frame of 558 bp which encodes 185 putative amino acids. Real-time PCR analysis revealed that the porcine APC10 mRNA expression shows a wide distribution and expression levels varies within a small different range in detected tissues. The deduced protein has a high identity with other eight species and comprises a conserved DOC domain. The phylogenetic tree indicated that porcine APC10 has the closest genetic relationship with human, monkey and dog. Promoter activity was demonstrated by transient transfection of 5'-deletion promoter/luciferase constructs into PK15 cells, and indicated that the proximal region from -2,052 to -1,764 is necessary for basal promoter activity. Positive cis-regulatory elements are present from -2,544 to -2,052 and from -3,114 to -2,774, while negative cis-regulatory elements may be present from -2,774 to -2,544. Yeast one-hybrid assay revealed Sp1 can interact with proximal promoter region of porcine APC10.

Molecular characterization and association analysis of FBXO40 with partial hematological indexes in pig.

F-box proteins are quite significant ubiquitin-proteasome pathway regulators in eukaryotic cells. FBXO40, a member of this large family, alters its expression pattern in muscle atrophy. Here we isolated most of the verified porcine FBXO40 coding sequence (CDS) (2258 bp) and assigned it to the porcine chromosome 13q4.1-4.6 by using the INRA-Minnesota porcine radiation hybrid panel, and we also explored the tissue expression distributions, which is relatively high in longissimus dorsi muscle, heart, low in kidney, small intestine, brain, hypophysis, lymphonode, thymus, spleen, large intestine, ovary, stomach, and undetectable in testis, liver, uterus and thyroid gland. Inferring phylogenetic tree was constructed to study the evolutionary implications. Moreover, a HindII (HincII)-RFLP (A/C) polymorphism in 3'-untranslated region (3'-UTR) of porcine FBXO40 gene was demonstrated by sequencing and PCR-restriction fragment length polymorphism (PCR-RFLP) analysis. Statistical analysis result of this polymorphism showed that the allele A was predominant in all detected indigenous breeds, but C in western introduced commercial breeds. The SNP was further analyzed in our experimental pig population including Tongcheng, Landrace, Large White, and crossbreds of Large White × (Landrace × Tongcheng) and Landrace × (Large White × Tongcheng). The association analysis results indicated that the A/C base substitution was associate with some hematological indexes, the hemoglobin concentration (P < 0.0001), mean corpuscular volume hemoglobin concentration (P = 0.0002) and mean corpuscular volume (P = 0.0138).

Expression and SNP association analysis of porcine FBXL4 gene.

As a kind of E3 ligase, the product of FBXL4 gene belongs to a member of FBLs which is the biggest eukaryotic subfamily of F-BOX proteins, it can recognize some substrate through particular protein-protein interaction domains. To investigate its functions, the polymorphism and association analysis was analyzed. The partial cDNA of porcine FBXL4 with 2384 bp long was first cloned; the deduced protein comprises a conserved F-BOX domain at position from the 277th to 332nd amino acid. The phylogenetic tree indicated porcine FBXL4 has the closest genetic relationship with bovine FBXL4 than other selected animal species. Ten tissue expression level of porcine FBXL4 mRNA fluctuated remarkably in a large range by quantitative RT-PCR analysis. For two identified SNPs, the genotyping analysis of Tail showed TT genotype owned dominance in introduced Landrace pig and miniature Guizhou and Wuzhishan breeds, but CC genotype was more than two other genotypes in miniature Laiwu breed. While in another genotyping analysis of BsaJI, CC genotype was obviously more than other genotypes in two kinds of Chinese miniature pig breeds and introduced Landrace pig breeds. Furthermore, the association analysis with immune traits and blood parameters revealed that SNP Tail was significantly associated with the lymphocyte percentage (P = 0.0166) and the antibody levels for pseudorabies virus vaccination (P = 0.0001) of neonate piglets at 0 day. Meanwhile, SNP BsaJI was significantly associated with lymphocyte percentage of individuals at 32 days (P = 0.0351), neutrophil percentage (P = 0.0005), the absolute lymphocyte count (P = 0.0458), and the mixed cells (P = 0.0010) of neonate piglets at 0 day.

Partial molecular characterization, expression pattern, polymorphism and association analysis of porcine SKP2 gene.

As a component of E3 ubiquitin protein ligases called SCFs, SKP2 protein belongs to a member of FBLs protein which is the biggest eukaryotic subfamily of F-BOX proteins with 12 members. In this study, we cloned and sequenced partial cDNA, intron 1 and intron 6 of porcine SKP2 gene. The partial cDNA is 1,402 bp long and has an open reading frame of 1,272 bp which encodes 424 putative amino acids. The deduced protein comprises a conserved F-BOX domain at position from the 90th to 140th amino acid. The phylogenetic tree indicated that porcine SKP2 has the closest genetic relationship with bovine SKP2 than other selected animal species. Quantitative RT-PCR analysis displayed that the tissue expression level of porcine SKP2 fluctuated remarkably in a large range, and it expressed in thymus with the highest level and in longissimus dorsi muscle with the lowest level. Two SNPs were identified, meanwhile, further polymorphism analysis with Cfr42I showed that AA genotype was in dominance absolutely among four kinds of unrelated Chinese indigenous miniature and one introduced Landrace pig breeds. In addition, association analysis with immune traits and blood parameters revealed that the SNP Cfr42I in intron 1 was significantly associated with red cell distribution width of neonate piglets at 0 day (P = 0.027).

Survey on the function, structure and operation of hospital ethics committees in Shanghai.

OBJECTIVE: The objectives of this study are to understand the current functions, structure and operation of hospital ethics committees (HECs) in Shanghai and to facilitate their improvement. METHODS: (1) A questionnaire survey, (2) interviews with secretaries and (3) on-site document reviews of HECs in Shanghai were used in the study, which surveyed 33 hospitals. RESULTS: In Shanghai, 57.56% of the surveyed hospitals established HECs from 1998 to 2005. Most HECs used bioethical review of research involving human subjects as well as bioethical review or consultation regarding medical care services and administrative decision- making. Of the surveyed HECs, 14.3% did not provide any formal bioethical training to the HECs' members and many HECs had no standard operating procedures. Some HECs had no clear definition of what was "conflict of interest" that should be considered by the HECs, while 44.4% of the HECs did not perform continuing review. DISCUSSION: After the issues of related national regulations, more and more hospitals established HECs in Shanghai, but the functions of HECs need to be further developed and formal training on bioethics should be provided to HEC members. To assure the independence and good performance of HECs, the conflict of interest procedure, the standard operating procedures and bioethical review should be improved. CONCLUSION: HECs in Shanghai had developed in the preceding 10 years and they played great roles in protecting the rights and welfare of human subjects and patients; some areas need improvement.

Effects of the preoperative administration of Yunnan Baiyao capsules on intraoperative blood loss in bimaxillary orthognathic surgery: a prospective, randomized, double-blind, placebo-controlled study.

Yunnan Baiyao is a well-known Chinese herbal medicine that has been used as a haemostatic drug for nearly 100 years. The aim of this clinical trial was to evaluate the efficacy and safety of Yunan Baiyao capsules on the reduction of blood loss in bimaxillary orthognathic surgery. 87 consecutive patients scheduled for simultaneous maxillary Le Fort I osteotomies and bilateral sagittal split ramus osteotomies (BSSRO) were enrolled in a prospective, randomized, double-blind, placebo-controlled clinical trial. Patients were administered Yunnan Baiyao capsules or placebo capsules, orally for 3 days before surgery. Intraoperative blood loss was estimated and the safety of Yunnan Baiyao capsules was evaluated. The total blood loss in the Yunnan Baiyao group (mean, 330.5+/-134.4 ml) was significantly lower than in the control group (mean, 420.3+/-175.9 ml). No allergic reactions, thromboembolic events or other side effects were recorded in this trial. It can be concluded that the preoperative use of Yunnan Baiyao capsules, in combination with hypotension anaesthesia, results in a reduction in intraoperative blood loss in bimaxillary orthognathic surgery. Yunnan Baiyao capsules are an effective and safe haemostatic Chinese medicine.

Full-length coding sequences, polymorphism and chromosomal localizations of the porcine EDG4 and EDG7 genes.

EDG4 and EDG7 are identified as cellular receptors for lysophosphatidic acid (LPA), belonging to the endothelial cell differentiation gene (EDG) family of G protein-coupled receptors (GPCR) which play an important role in the function of LPA. In this study, we presented the complete coding sequences of porcine EDG4 and EDG7 genes. The nucleotide sequences and the predicted protein sequences share high sequence identity with other mammals. Spatial expression analysis by reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that porcine EDG4 and EDG7 genes are mainly expressed in brain, liver, spleen, lung, kidney, large intestine, small intestine, but absent in muscle tissues. Radiation hybrid mapping data indicated that EDG4 and EDG7 map to q2.1 of pig chromosome 2 (SSC2) and q2.6-3.2 of pig chromosome6 (SSC6), respectively. A T/C single nucleotide polymorphism (SNP) in the coding sequence of porcine EDG4 was identified. A PCR-restriction fragment length polymorphism (PCR-RFLP) method was employed to genotype this locus among Guizhou Miniature, Guangxi Miniature, Laiwu, Wuzhishan, Tongcheng, Landrace and Yorkshire pigs. The association analysis suggested that the EDG4 genotype was associated with carcass length (P < 0.05) and drip loss percentage (P < 0.05) in the experimental population consisting of Tongcheng, Landrace, Yorkshire and two crossbred porcine populations (Wang et al. Biochem Genet (1-2):51-62, 2007).

Partial molecular cloning, characterization, and analysis of the subcellular localization and expression patterns of the porcine OTUB1 gene.

The product of the OTUB1 gene is a member of the OTU superfamily of predicted cysteine proteases and inhibits cytokine gene transcription via its interaction with a ubiquitin protease and E3 ubiquitin ligase. To further understand the functions of the porcine OTUB1 gene, the subcellular localization of porcine OTUB1 protein was analyzed. We first cloned a partial DNA sequence of porcine OTUB1 which contained an 816 bp ORF encoding 271 amino acids. The deduced protein product was found to contain an OTU domain. The corresponding porcine OTUB1 protein was subsequently demonstrated to localize predominantly in the nucleus by confocal fluorescence microscopy. By spatial expression analysis, we further found that OTUB1 is highly expressed in the brain, liver, spleen, lung, kidney, large intestine, small intestine, stomach, ovary, uterus and thymus. In contrast, only low levels of this gene were evident in the heart, dorsal muscles and leg muscle of the pig. This is the first report to show the subcellular localization of porcine OTUB1, and our current data provides us with an important basis for conducting further studies on the functions and regulatory mechanisms underlying the role of OTUB1 gene in the immune system.

Sequence characterization, polymorphism, and chromosomal localizations of the porcine CapZ genes.

CapZ is a widely distributed and highly conserved actin-binding protein that caps the barbed end of actin filaments and nucleates actin polymerization in a Ca(2+)-independent manner. In myofibrils, it is localized in the Z-lines. In this study, we cloned and characterized Capz subunit genes from the pig muscle. The nucleotide sequences and the predicted protein sequences share high sequence identity with other mammalian orthologs. The reverse transcriptase polymerase chain reaction (RT-PCR) revealed that porcine Capzbeta, Capzalpha1, and Capzalpha2 genes are expressed in all 11 tissues studied (liver, spleen, small intestine, large intestine, lymph node, kidney, heart, skeletal muscle, brain, fat, and lung) but in variable amounts. Radiation hybrid mapping data indicated that Capzbeta, Capzalpha1, and Capzalpha2 map to q2.1-2.6 of pig chromosome 6 (SSC6), q1.6-q2.2 of pig chromosome 4 (SSC4), and q1.3-2.3 of pig chromosome 18 (SSC18), respectively. An A/C single nucleotide polymorphism in Capzbeta intron 4 was identified with a HhaI PCR restriction fragment length polymorphism, which showed great allele frequency differences between Guizhou Xiang, Guangxi Bama, Wuzhishan, Tongcheng, Landrace, and Yorkshire pigs. The association analysis suggested that the Capzbeta genotype was associated with leaf fat (P < 0.05) in our experimental population.

Prolonged gene expression in mouse lung endothelial cells following transfection with Epstein-Barr virus-based episomal plasmid.

The development of a strategy to deliver a gene to pulmonary endothelium will be useful for gene function study and for pulmonary gene therapy. Cationic lipidic vectors are efficient in gene transfer to pulmonary endothelium via the vascular route; however, gene expression is transient and lasts for only a few days. In this study, we show that pulmonary gene transfer via cationic lipidic vectors can be significantly improved using an Epstein-Barr virus (EBV)-based expression plasmid. Systemic administration of cationic liposomes followed by the EBV-based plasmid led to gene expression in the lung that lasted for more than 3 weeks. Prolonged and high levels of gene expression can also be obtained in primary mouse lung endothelial cells (MLEC) following lipofection with an EBV-based plasmid. These results suggest the utility of this gene transfer protocol in studying the expression of cloned genes in lung endothelial cells and in pulmonary gene therapy.

Role of zinc in pulmonary endothelial cell response to oxidative stress.

Although zinc is a well-known inhibitor of apoptosis, it may contribute to oxidative stress-induced necrosis. We noted that N,N,N',N'- tetrakis(2-pyridylmethyl)ethylenediamine (TPEN; >10 microM), a zinc chelator, quenched fluorescence of the zinc-specific fluorophore Zinquin and resulted in an increase in spontaneous apoptosis in cultured sheep pulmonary artery endothelial cells (SPAECs). Addition of exogenous zinc (in the presence of pyrithione, a zinc ionophore) to the medium of SPAECs caused an increase in Zinquin fluorescence and was associated with a concentration-dependent increase in necrotic cell death. Exposure of SPAECs to TPEN (10 microM) resulted in enhanced apoptosis after lipopolysaccharide or complete inhibition of t-butyl hydroperoxide (tBH)-induced necrosis. We further investigated the role of two zinc-dependent enzymes, poly(ADP-ribose) polymerase (PARP) and protein kinase (PK) C, in tBH toxicity. tBH toxicity was only affected by the PARP inhibitors 4-amino-1,8-naphthalimide or 3-aminobenzamide over a narrow range, whereas the PKC inhibitors bisindolylmaleimide and staurosporine significantly reduced tBH toxicity. tBH caused translocation of PKC to the plasma membrane of SPAECs that was partially inhibited by TPEN. Thus pulmonary endothelial cell zinc inhibits spontaneous and lipopolysaccharide-dependent apoptosis but contributes to tBH-induced necrosis, in part, via a PKC-dependent pathway.