METHODS AND RESULTS: In this study, we have developed a simplified, tissue culture-independent protocol to deliver the CRISPR/Cas9 system through in planta transformation in Malaysian rice (Oryza sativa L. subsp. indica cv. MR 219). Sprouting seeds with cut coleoptile were used as the target for the infiltration by Agrobacterium tumefaciens and we achieved 9% transformation efficiency. In brief, the dehusked seeds were surface-sterilised and imbibed, and the coleoptile was cut to expose the apical meristem. Subsequently, the cut coleoptile was inoculated with A. tumefaciens strain EHA105 harbouring CRISPR/Cas9 expression vector. The co-cultivation was conducted for five to six days in a dark room (25 ± 2 °C) followed by rooting, acclimatisation, and growing phases. Two-month-old plant leaves were then subjected to a hygromycin selection, and hygromycin-resistant plants were identified as putative transformants. Further validation through the polymerase chain reaction verified the integration of the Cas9 gene in four putative T0 lines. During the fruiting stage, it was confirmed that the Cas9 gene was still present in three randomly selected tillers from two 4-month-old transformed plants.

METHODS: A cross-sectional study was conducted in 2 community health clinics within the Gombak district between September and December 2017. Adults with T2DM were selected via systematic random sampling and screened using the Neuropathy Symptoms Score (NSS). Clinical records of participants' foot examinations were reviewed to identify positive findings of DPN and compared with the NSS.

RESULTS: The study's sample comprised 425 patients. Most had co-morbidities, including hypertension, dyslipidaemia and pre-existing DM-related complications. About two-thirds of them performed no daily foot inspection and had no proper footwear. The proportion of patients with positive NSS was 49.4%. However, only 0.2% were diagnosed with positive DPN in their clinical foot examination record.

MATERIALS AND METHODS: This was a 12-month randomized controlled trial, in which adults with a controlled attenuation parameter (CAP) score >250 dB/m and a fibrosis score <10 kPa were randomly assigned to receive a standardized P. niruri extract at a dose of 3,000 mg daily (n = 112) or a placebo (n = 114). The primary outcomes were changes in CAP score and liver enzyme levels, while the secondary outcomes were changes in other metabolic parameters. The analysis was performed on an intention-to-treat basis.

RESULTS: After 12 months, there was no significant difference in the change of CAP score between the intervention and control groups (-15.05 ± 36.76 dB/m vs. -14.74 ± 41.08 dB/m; p = 0.869). There was also no significant difference in the changes of liver enzyme levels between the two groups. However, the intervention group showed a significant reduction in fibrosis score, which was not observed in the control group (-0.64 ± 1.66 kPa versus 0.10 ± 1.61 kPa; p = 0.001). No major adverse events were reported in either group.