Spatially resolved transcriptomic profiling of degraded and challenging fresh frozen samples.

Spatially resolved transcriptomics has enabled precise genome-wide mRNA expression profiling within tissue sections. The performance of methods targeting the polyA tails of mRNA relies on the availability of specimens with high RNA quality. Moreover, the high cost of currently available spatial resolved transcriptomics assays requires a careful sample screening process to increase the chance of obtaining high-quality data. Indeed, the upfront analysis of RNA quality can show considerable variability due to sample handling, storage, and/or intrinsic factors. We present RNA-Rescue Spatial Transcriptomics (RRST), a workflow designed to improve mRNA recovery from fresh frozen specimens with moderate to low RNA quality. First, we provide a benchmark of RRST against the standard Visium spatial gene expression protocol on high RNA quality samples represented by mouse brain and prostate cancer samples. Then, we test the RRST protocol on tissue sections collected from five challenging tissue types, including human lung, colon, small intestine, pediatric brain tumor, and mouse bone/cartilage. In total, we analyze 52 tissue sections and demonstrate that RRST is a versatile, powerful, and reproducible protocol for fresh frozen specimens of different qualities and origins.

A Systematic Review of the Diagnostic Criteria Used to Select Participants in Randomised Controlled Trials of Interventions Used to Treat Cervicogenic Headache.

OBJECTIVE: The aim of this study was to determine the diagnostic criteria used in randomized controlled trials to define trial participants as having cervicogenic headache (CeH). BACKGROUND: While animal and human studies suggest a biological basis for "cervicogenic" headaches the diagnostic criteria necessary to evidence CeH are debated. METHODS: A systematic review was undertaken guided by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. An electronic search of MEDLINE, Cochrane, CINHAL, Pedro, AMED, and EMBASE online databases of randomized controlled trials published between January 1983 and October 2018 found 39 randomized controlled trials which met the study inclusion criteria. RESULTS: Two independent reviewers found most trials cited 1 (31/39; 79.5%) or more (3/39; 7.6%) references to define the criteria used to identify CeH in their study participants. In spite of updated publications concerning the characteristics and definition of CeH, many (27/39; 69.2%) used diagnostic criteria published between 5 and 24 years prior to the randomized controlled trial. The most commonly cited diagnostic criteria included unilateral headache (18/39; 46.2%), cervical movement or sustained posture that either provoked (18/39; 46.2%) or precipitated (17/39; 43.6%) the headache. Fifteen trials did not exclude participants with signs or symptoms of other forms of headache. Although anesthetic blockade of cervical tissue or nerves is considered necessary for a "definitive" diagnosis, only 7.6% (3/39) of trials used anesthetic blockade at recruitment. CONCLUSIONS: This systematic review evidences the heterogeneity in the clinical characteristics used to diagnose CeH in participants recruited in randomized controlled trials. It raises a significant concern about the usefulness of currently available randomized controlled trials to determine the clinical merits of the treatment and management of people with CeHs. Our systematic review suggests that most randomized controlled trials published to date have investigated headaches with a clinical presentation involving the neck that maybe better defined as "possible," "probable," or "definitive" CeH depending on how well the diagnostic criteria used align with the most recent edition (3rd) of the International Classification of Headache Disorders.

Postherpetic Neuralgia: Practical Experiences Return to Traditional Chinese Medicine.

Post-herpetic neuralgia (PHN) is a complication of herpes zoster that can cause different types of pain in the affected area. It often occurs mainly in severe cases of herpes zoster. The problem is defined as a persisting pain for 90-120 days after relieving of acute phase of herpes lesions. This complication causes suffering in patients and reduces the quality of life. In western medicine's viewpoints, PHN is due to disturbance in local and dermatomal nerves. There are several topical and systemic drugs that are used to manage the pain relief. In traditional medicine (TM), PHN is mostly due to incomplete heat and damp clearing in liver and spleen meridians, qi and toxic pathogens stagnation, accumulation of yin (blood stagnation in microcapillary), internal fire, and heat and obstruction of meridians. Acupuncture works based on the eradication of wind, clearing of heat, and destroying of damp by regulating qi and blood movement. In clinics, several methods of TM are used to relief PHN, such as simultaneous needling, surrounding needling, acupuncture, electro acupuncture, moxibustion, wet cupping or hijamat, and herbal medicine. In this review, we discussed all these methods and their role in reducing PHN and pain.