Transcutaneous nerve stimulation (TENS) treatment was given for 30 min to 37 patients divided into 3 groups of 10 patients and 1 group of 7 patients. Two groups received low-frequency (2 Hz) and the other 2 groups high-frequency (100 Hz) stimulation. A diagnostic lumbar cerebrospinal fluid (CSF) sample was obtained immediately before and after stimulation. The CSF samples were subjected to analysis of immunoreactive (ir) opioid peptides, Met-enkephalin-Arg-Phe (MEAP) from preproenkephalin and dynorphin A (Dyn A) from preprodynorphin, respectively. Low frequency TENS applied on the hand and the leg resulted in a marked increase (367%, P less than 0.05) of ir-MEAP but not ir-Dyn A, whereas high-frequency (100 Hz) TENS produced a 49% increase in ir-Dyn A (P less than 0.01) but not ir-MEAP. This is the first report in humans that 2 Hz and 100 Hz peripheral stimulation induces differential release of peptides from preproenkephalin and preprodynorphin, respectively.

There is an urgent clinical need for safe and effective treatment agents and therapy targets for estrogen receptor negative (ER−) breast cancer. G protein-coupled receptor 30 (GPR30), which mediates non-genomic signaling of estrogen to regulate cell growth, is highly expressed in ER− breast cancer cells. We here showed that activation of GPR30 by the receptor-specific agonist G-1 inhibited the growth of ER− breast cancer cells in vitro. Treatment of ER− breast cancer cells with G-1 resulted in G2/M-phase arrest, downregulation of G2-checkpoint regulator cyclin B, and induction of mitochondrial-related apoptosis. The G-1 treatment increased expression of p53 and its phosphorylation levels at Serine 15, promoted its nuclear translocation, and inhibited its ubiquitylation, which mediated the growth arrest effects on cell proliferation. Further, the G-1 induced sustained activation and nuclear translocation of ERK1/2, which was mediated by GPR30/epidermal growth factor receptor (EGFR) signals, also mediated its inhibition effects of G-1. With extensive use of siRNA-knockdown experiments and inhibitors, we found that upregulation of p21 by the cross-talk of GPR30/EGFR and p53 was also involved in G-1-induced cell growth arrest. In vivo experiments showed that G-1 treatment significantly suppressed the growth of SkBr3 xenograft tumors and increased the survival rate, associated with proliferation suppression and upregulation of p53, p21 while downregulation of cyclin B. The discovery of multiple signal pathways mediated the suppression effects of G-1 makes it a promising candidate drug and lays the foundation for future development of GPR30-based therapies for ER− breast cancer treatment.

BackgroundSalivary gland ultrasonography (SGUS) is promising on diagnosis and disease monitoring of primary Sjögren’s syndrome (pSS). However, there is neither study concerning the association between SGUS and salivary function in pSS.ObjectivesThis study aimed to explore how SGUS features reflected the secretory function of salivary glands in pSS.MethodsThe subjects who presented with ocular dryness and/or oral dryness were recruited from rom July 2021 to July 2022 at our department. The subjects were classified as pSS and non-SS according to the 2002 the American-European Consensus Group (AECG) criteria for pSS. Unstimulated whole salivary flow (UWSF) and stimulated whole salivary flow (SWSF) were assessed to evaluate salivary function. SGUS was performed to bilateral parotid and submandibular glands. Each gland was assessed by a well-accepted 0–4 scoring system (gland score) and a 0-3 scoring system from the Outcome Measures in Rheumatology Clinical Trials working group (OMERACT score).Results① Among 309 enrolled subjects with suspected pSS, 95 subjects were excluded due to being classified as secondary SS, or failing to receive the designated SGUS examination or labial salivary glands biopsy. Therefore, 214 subjects were qualified for statistical analysis, including 116 pSS patients. The mean age of pSS patients was 47.1±13.3 years old and 97% of them were female, age-match and sex-match to the non-SS controls.② Compared to the pSS patients without hypofunction, pSS patients with salivary hypofunction (n=71, SWSF ≤0.7mL/min) were significantly older, higher positive rate of the Van Bijsterveld score, higher positive rate of anti-SSA/Ro, higher incidence of UWSF≤0.1mL/min and lower incidence of thrombocytopenia (allp<0.05).③ The pSS patients with salivary hypofunction had significantly higher total OMERACT score (10 [interquartile range (IQR), 8-12] vs. 8 [IQR,6-8],p<0.001), and total gland score (12 [IQR,10-12] vs. 8 [IQR,6-11],p<0.001) than the patients without salivary hypofunction (Figure 1A). ROC curve confirmed total OMERACT score (AUC: 0.804, 95%CI: 0.727-0.881,p<0.001) and total gland score (AUC: 0.792, 95%CI: 0.704-0.881,p<0.001) could significantly distinguish pSS patients with salivary hypofunction from those without salivary hypofunction. The cut-off values of total OMERACT score were 7 and 9, and of total gland score were 9 and 11, according to the Youden index. The absolute value of SWSF in the pSS patients with total gland score >11 was significantly lower than those with total gland score of 9~11 or total gland score <9 (bothp<0.05). The absolute value of SWSF in the pSS patients with total OMERACT score >9 was significantly lower than those with total gland score of 7~9 or total gland score <7 (both p<0.001, Figure 1B).④ Total OMERACT score and total gland score were combined to generate a 3×3 matrix model and then the estimated risk probability for salivary hypofunction of pSS was calculated in each grid (Figure 1C). Accordingly, pSS patients could be stratified into three risk subgroups: (i) High-risk (n=39): pSS patients whose total OMERACT score was >9 and total gland score ≥9 had estimated risk probability greater than 80%, and the actual incidence of SWSF≤0.7mL/min was 95% (37/39). (ii) Low-risk (n=28): pSS patients whose total OMERACT score was <7; or whose total OMERACT score was 7~9 and total gland score <9 had estimated risk probability less than 35%, and the actual incidence of SWSF≤0.7mL/min was 14% (4/28). (iii) Moderate-risk (n=49): pSS patients who fulfilled neither high-risk nor low-risk had an actual incidence of SWSF≤0.7mL/min in a moderate level (61%, 30/49).ConclusionThis study first reported B-mode ultrasonography examination on four major salivary glands could reflect salivary function of pSS. We also built a matrix risk model composed of SGSU gland score and OMERACT score associated with salivary hypofunction in pSS which was conviently used for rheumatologists, especially in specific clinical situations that SWSF was not available.REFERENCES:NIL.Acknowledgements:NIL.Disclosure of InterestsNone Declared.