Ziziphus

A total of 18 batches of HQT were included in the present study. Four batches coded as PHY906-6, 7, 8, 10 were manufactured with PhytoCeutica's proprietary SOP. Eight batches of HQT were purchased from Sun Ten Pharmaceutical Co. LTD in Taiwan and designated as HQT-E, F, G, H, I, J, K and L. Six batches of HQT were obtained from various vendors (Chung Song Zong, Ko Da, Min Tong, Sheng Chang, Sheng Foong, Kaiser; Taiwan) who did not provide quality information, and were labeled as HQT-CSZ, KD, MT, SC, SF and KP3. The proprietary standard operating procedures (SOP) by PhytoCeutica for PHY906 used hot water extraction (80°C) of four herbs, namely Scutellaria baicalensis Georgi (S), Paeonia lactiflora Pall. (P), Glycyrrhiza uralensis Fisch. (G) and Ziziphus jujuba Mill. (Z) (ratio 3:2:2:2). The hot water extraction is then spray dried with insoluble dextran into a granulated powder, packaged and stored in foil containers at 4°C.
Chemical standards including baicalin (S), baicalein (S), wogonin (S), scutellarin (S), glycyrrhizin (G), ononin (G), liquiritin (G), liqiritigenin (G), paeoniflorin (P) and albiflorin (P), were obtained from Chromadex (USA). Apigenin and formic acid were obtained from Sigma-Aldrich (USA). Solvents were of LC/MS grade from JT Baker (USA).

Gir Protected Area (PA) [1,883 km², 20°57′ to 21°20′ N latitude and 70°27′ to 71°13′ E longitude] is a dry deciduous forest [32] situated in Gujarat province, western India (Fig. 1) and is made up of a Sanctuary (with human settlements and regulated grazing and other rights; [24] ) covering 1,153 km², a 259 km² National Park (devoid of humans) and 471 km² of additional reserve, protected and unclassified forests. Gir PA has a semi-arid climate with an average minimum and maximum temperature ranging from 5° to 38°C and an average rainfall of 980 mm [31] . Rugged hilly terrains form the catchments of seven perennial rivers. Dominant vegetation included Tectona grandis, Anogeissus spp, Acacia spp and Ziziphus spp.
Gir has a diverse assemblage of wild fauna. Apart from the last free-ranging population of the Asiatic lion, some of the other carnivores are leopard (Panthera pardus), striped hyena (Hyaena hyaena), jackal (Canis aureus) and ratel (Mellivora capensis). Major wild prey species of lions were chital (Axis axis), sambar (Rusa unicolor), nilgai (Boselaphus tragocamelus) and wild pig (Sus scrofa) [31] .
Gir Protected Area has 50 Maldhari settlements (nesses). A ness consists of a cluster of thatch and mud hutments of 3–20 Maldhari families. [11] , [31] , [33] . Each Maldhari family rears about 20–100 regionally famous indigenous breed of livestock, primarily Jafrabadi breed of buffalo (Bubalus bubalis) and Gir breed of cattle (Bos indicus). Often one or two camels (Camelus dromidarius) are kept for carrying fuel wood and fodder. The sale of dairy products has always been the mainstay of Maldharis’ traditional economy [33] . Our study area covered the livestock grazing areas of a cluster of six nesses namely Asundrali, Dodhi, Gudjinjva, Khajuri, Leriya and Mindha (Fig. 1) which represent a typical scenario across Gir PA.

The genome sequences of Ziziphus jujuba Mill. were obtained from the National Center for Biotechnology Information (NCBI) database [19 (link)]. The sequences of 14, 9 identified MdRNases, PyRNases were downloaded from the National Center for Biotechnology Information (NCBI) database. In addition, the sequences of8 identified OsRNases were downloaded [1 (link)]. RNases were identified by two rounds of BLASTP searches. First, the sequences of all OsRNases were used to search for possible ZjRNases sequences via TBtools [36 (link)]. Then, NCBI Batch CD-Search was used to confirm whether the candidate RNases contained the RNase_T2 superfamily domain (pfam00445) or the Ribonuclease_T2 domain (cl00208). A total of 4 ZjRNase genes were ultimately identified in the genome. The protein length, isoelectric point (pI) and molecular weight (MW) were subsequently predicted.

A prospective, randomized clinical trial was conducted from March 2020 to December 2021 (Clinical Trials Registration ID: NCT04831112) to compare the effects of dressing with EUSOL or honey for managing NF wounds in the emergency department after approval from Ethics Review Committee (IRB-1491/DUHS/Approval/2020). After giving their informed consent, patients were assigned to one of the two groups via the lottery method in a single-blinded procedure. Investigators were blinded to the procedure. The manuscript follows the CONSORT guidelines for all analyses and the randomized trial reporting specified in advance [8 ].
Inclusion criteria included adults (>18 years) with a diagnosis of NF with the broadest span of wound <20 cm and who underwent surgical debridement followed by dressing were enrolled in the study.
Exclusion criteria included diabetic wounds, traumatic wounds, and patients who expired during the postoperative period.
Intervention
This study included patients with necrotizing soft tissue infection admitted via the emergency department and underwent thorough evaluation, including history, examination, and investigation. Following admission and after the stabilization of vital signs with intravenous fluids and empiric antibiotics, all patients underwent surgical debridement. All patients are given daily routine dressings following debridement. A total of 175 patients enrolled in the study, with 90 in the intervention group (honey) and 85 in the control group (EUSOL). A sample of infected tissue during surgical debridement was taken for pus culture and sensitivity and tissue for histopathology. The wound of all patients healed by secondary intention. The wound was covered with a honey layer of 4 ml per square inch, followed by a dressing covered by Opsite, which was changed every 24 hours. During the dressing change, clearance of slough and appearance of granulation were assessed. Ber Honey (Ziziphus mauritiana) was collected from the district of Tharparkar Pakistan and sterilized before being used for dressing. Gamma radiation is used to sterilize the honey and get rid of the bacterium spores known to reside in raw honey.
Data collection
The dataset includes the baseline demographic factors including age, gender, comorbidities including diabetes mellitus, ischemic heart disease, chronic liver disease, chronic kidney disease, hypertension, site involved by disease, duration of symptoms, and smoking. The second section contains outcome factors, including days needed for clearance of slough and appearance of granulation tissue, wound healing time, length of hospital stays, and patient satisfaction.
Statistical analysis
SPSS v.25 was used to analyze the data. Statistics were considered significant for a p-value less than 0.05. Categorical data were analyzed as proportions, while continuous parametric data were analyzed as mean with standard deviation. Chi-square was used to measure categorical variables, whereas continuous parametric variables’ variance was assessed using the student’s t-test.