ThermomiR-377-3p-induced suppression of Cirbp expression is required for effective elimination of cancer cells and cancer stem-like cells by hyperthermia.

BACKGROUND: In recent years, the development of adjunctive therapeutic hyperthermia for cancer therapy has received considerable attention. However, the mechanisms underlying hyperthermia resistance are still poorly understood. In this study, we investigated the roles of cold­inducible RNA binding protein (Cirbp) in regulating hyperthermia resistance and underlying mechanisms in nasopharyngeal carcinoma (NPC). METHODS: CCK-8 assay, colony formation assay, tumor sphere formation assay, qRT-PCR, Western blot were employed to examine the effects of hyperthermia (HT), HT + oridonin(Ori) or HT + radiotherapy (RT) on the proliferation and stemness of NPC cells. RNA sequencing was applied to gain differentially expressed genes upon hyperthermia. Gain-of-function and loss-of-function experiments were used to evaluate the effects of RNAi-mediated Cirbp silencing or Cirbp overexpression on the sensitivity or resistance of NPC cells and cancer stem-like cells to hyperthermia by CCK-8 assay, colony formation assay, tumorsphere formation assay and apoptosis assay, and in subcutaneous xenograft animal model. miRNA transient transfection and luciferase reporter assay were used to demonstrate that Cirbp is a direct target of miR-377-3p. The phosphorylation levels of key members in ATM-Chk2 and ATR-Chk1 pathways were detected by Western blot. RESULTS: Our results firstly revealed that hyperthermia significantly attenuated the stemness of NPC cells, while combination treatment of hyperthermia and oridonin dramatically increased the killing effect on NPC cells and cancer stem cell (CSC)­like population. Moreover, hyperthermia substantially improved the sensitivity of radiation­resistant NPC cells and CSC­like cells to radiotherapy. Hyperthermia noticeably suppressed Cirbp expression in NPC cells and xenograft tumor tissues. Furthermore, Cirbp inhibition remarkably boosted anti­tumor­killing activity of hyperthermia against NPC cells and CSC­like cells, whereas ectopic expression of Cirbp compromised tumor­killing effect of hyperthermia on these cells, indicating that Cirbp overexpression induces hyperthermia resistance. ThermomiR-377-3p improved the sensitivity of NPC cells and CSC­like cells to hyperthermia in vitro by directly suppressing Cirbp expression. More importantly, our results displayed the significantly boosted sensitization of tumor xenografts to hyperthermia by Cirbp silencing in vivo, but ectopic expression of Cirbp almost completely counteracted hyperthermia-mediated tumor cell-killing effect against tumor xenografts in vivo. Mechanistically, Cirbp silencing-induced inhibition of DNA damage repair by inactivating ATM-Chk2 and ATR-Chk1 pathways, decrease in stemness and increase in cell death contributed to hyperthermic sensitization; conversely, Cirbp overexpression-induced promotion of DNA damage repair, increase in stemness and decrease in cell apoptosis contributed to hyperthermia resistance. CONCLUSION: Taken together, these findings reveal a previously unrecognized role for Cirbp in positively regulating hyperthermia resistance and suggest that thermomiR-377-3p and its target gene Cirbp represent promising targets for therapeutic hyperthermia.

Covalent immobilization of tyrosinase on magnetic multi-walled carbon nanotubes for inhibitor screening.

The inhibition of tyrosinase is considered to be a common therapeutic strategy for some hyperpigmentation disorders. Screening of tyrosinase inhibitors is of great significance to the treatment of pigmentation diseases. In this study, tyrosinase was covalently immobilized on magnetic multi-walled carbon nanotubes for the first time, and the immobilized tyrosinase was applied for ligand fishing of tyrosinase inhibitors from complex medicinal plants. The immobilized tyrosinase was characterized by transmission electron microscopy, atomic force microscopy, Fourier-transform infrared spectroscopy, vibrating sample magnetometry, and thermo-gravimetric analyzer, which indicated that tyrosinase was immobilized onto magnetic multi-walled carbon nanotubes. The immobilized tyrosinase showed better thermal stability and reusability than the free one. The ligand was fished out from Radix Paeoniae Alba and identified as 1,2,3,4,6-pentagalloylglucose by ultra-performance liquid chromatography-quadrupole time-of-flight high-resolution mass spectrometry. 1,2,3,4,6-pentagalloylglucose was found to be a tyrosinase inhibitor with similar half maximal inhibitory concentration values of 57.13 ± 0.91 µM compared to kojic acid (41.96 ± 0.78 µM). This work not only established a new method for screening tyrosinase inhibitors but also holds considerable potential for exploring the new medicinal value of medicinal plants.

[Clinical effect of different schemes of mild moxibustion for treatment of knee osteoarthritis].

OBJECTIVE: To observe the therapeutic effect of different schemes of mild moxibustion for treatment of (yang-deficiency induced cold-accumulation type)knee osteoarthritis (KOA). METHODS: Fifty-nine KOA patients were chosen and randomly divided into control group (n =30) and test group (n =29). Patients of the control group were treated by mild moxi- bustion of Neixiyan (EX-HE4) and Waixiyan (ST 35) for 30 min, once daily for 14 days (two courses), and those of the test group were treated by mild moxibustion of EX-HE 4 and ST 35 for 30 mini once daily for 7 days (the same to control group), followed by moxibustion of Yaoyangguan (GV 3) and Mingmen (GV 4) once daily for next 7 days more. In addition, patients of the two groups were also treated by routine acupuncture stimulation of EX-HE 4, ST 35, Yanglingquan (GB 34), Kunlun (BL 60), etc. The interval between two therapeutic courses was one day. The Lysholm Knee Score Scale (LKSS) was used to evaluate the therapeutic effect. Visual analogue scale (VAS) was employed to assess the patient's knee-joint pain severity (arthralgia), and scores of morning stiffness, arthrocele, and walking restraint degree of the knee-joint were also evaluated before and after the treatment. RESULTS: After the treatment, the scores of VAS, morning stiffness, arthrocele and walking restraint degree of the knee-joint of both groups were significantly decreased (P<0. 05), and the scores of the test group were obviously lower than those of the con- trol group (P<0. 05). The effective rate of the test group was 89.66% (26/29)which was obviously higher than that (70.00%, 21/30) of the control group (P<0. 05). CONCLUSION: Mild moxibustion of Neixiyan (EX-HE 4) and Waixiyan (ST 35, local acu- points), and Yaoyangguan (GV 3) and Mingmen (GV 4) has a better therapeutic effect for KOA patients than moxibustion of local acupoints only.

[Efficacy comparison of lumber disc herniation treated with mild moxibustion at Yaoyangguan (GV 3) under different conditions].

OBJECTIVE: To compare the difference in clinical efficacy on lumber disc herniation (LDH) treated with Yaoyangguan (GV 3) between mild moxibustion under thermosensitive condition and that under non-thermo-sensitive condition. METHODS: Fifty-seven LDH patients were selected as the study objects. Mild moxibustion at Yaoyangguan (GV 3) was applied for 45 min each time. Additionally, the conventional acupuncture was given, once a day, for 20 days. At the end of treatment, two groups were classified. A thermosensitive acupoint group (thermosensitive group) was composed of the cases with thermo-sensitization at Yaoyangguan (GV 3) and presenting for > or =4 times in the entire treatment. A non-thermosensitive acupoint group (tranquilization group) was composed of the cases without thermo-sensitization or the frequency of thermo-sensitization <4 times in the entire treatment. The modified Japanese orthopedics association scoring system (M-JOA) was adopted to observe the cases before and after treatment and 6 months after treatment in the two groups. The efficacy was compared between the two groups. RESULTS: After treatment and in 6 months after treatment, the score of M-JOA was apparently reduced as compared with that before treatment in the two groups (all P<0.05). The score reducing in the thermosensitization group was more obvious than that in the tranquilization group (both P<0.01). After treatment and in 6 months after treatment, the curative rate and remarkably effective rate were 89.7% (26/29) and 79.3% (23/29), which was better than 71.4% (20/28) and 60.7% (17/28, both P<0.05) in the tranquilization group separately. CONCLUSION: Mild moxibustion at acupoint under thermosensitive condition achieves the better short-term and long-term effects as compared with that under non-thermosensitive condition.

[Relationship between patients' sensitivity to moxibustion stimulation and clinical effect in the treatment of peripheral facial palsy].

OBJECTIVE: To compare the outcome difference between the heat-sensitive and non-sensitive moxibustion stimulation of Yifeng (TE 17) in the treatment of peripheral facial palsy. METHODS: A total of 43 patients with peripheral facial paralysis were divided into heat-sensitive moxibustion (n = 21) and non-sensitive moxibustion (n = 22) groups in accordance with their reactions to moxa-heat stimulation. Mild moxibustion was applied to bilateral Yifeng (TE 17) for 45 minutes, followed by mild acupuncture stimulation of Cuanzhu (BL 2), Yangbai (GB 14), Sibai (ST 2), Hegu (LI 4), etc. Moxibustion treatment was conducted once daily for 10 days, and acupuncture treatment given once daily for 20 days (with 2 days interval between every 10 days). Following moxibustion, if the patient felt regional heat penetrating to the deep tissue, extending peripherally, or propagating to other part of the body, or felt mild warm in the stimulated region but warmer in the slightly distant part, or felt mild warm on the skin surface but warmer in the deep tissue, it was considered to be heat-sensitivity. Patients with occurrence of heat-sensitivity being equal to and more than 3 times during the 10 sessions of treatment were assigned to heat-sensitive group, and those with occurrence of heat-sensitivity being equal to or less than 2 times assigned to non-sensitive moxibustion group. According to Portmann Scale (including movement and resting posture) for the voluntary movement state of the face, forehead, winkles eye closure, open mouth smile, snarl, and pucker; points 0, 1, 2 and 3 indicate no muscular movement, marked asynersis, asynersis and normal, respectively. In the light of resting posture of the eye, the nasolabial fold and mouth corner, 0, 1, and 2 points indicate severe, mild dissymmetry and normal, separately. In evaluation of the therapeutic effect, twenty points (at most) indicate cured, 17-19 points marked improvement; 14- 16 points improved, and < or = 13 points failure. RESULTS: Compared with pre-treatment, Portmann scores of both heat-sensitive moxibustion and non-sensitive moxibustion groups were significantly increased after the treatment (P < 0.001), and the score of the heat-sensitive moxibustion group was markedly higher than that of the non-sensitive moxibustion group (P < 0.05). Of the 21 and 22 facial palsy patients in the heat-sensitive moxibustion and non-sensitive moxibustion groups, 8 and 5 cases were cured, 10 and 7 were markedly effective, 3 and 10 were improved, with the markedly effective rates being 85.71% and 54.55%, respectively. The therapeutic effect of the heat-sensitive moxibustion group was statistically better (P < 0.05). CONCLUSION: Heat-sensitive moxibustion is significantly superior to non-sensitive moxibustion in improving symptoms of peripheral facial palsy patients, suggesting a necessity for paying attention to patients' reactions during moxibustion treatment.

Distribution, source apportionment, and transport of PAHs in sediments from the Pearl River Delta and the Northern South China Sea.

Polycyclic aromatic hydrocarbons (PAHs) were measured in 59 surface sediments from rivers in the Pearl River Delta and the northern continental shelf of the South China Sea. Total PAH concentrations varied from 138 to 6,793 ng/g dry weight. The sources of PAH inputs to sediments in the Pearl River Delta were qualitatively and quantitatively determined by diagnostic ratios and principal components analysis with multiple linear regression. The results showed that on average coal and wood combustion, petroleum spills, vehicle emissions, and nature sources contributed 36%, 27%, 25%, and 12% of total PAHs, respectively. Coal and biomass combustion was the main source of PAHs in sediments of the South China Sea, whereas petroleum combustion was the main source of pyrolytic PAHs in riverine and estuarine sediments of the Pearl River Delta. Perylene was formed in situ in river sediments and then transported to coastal areas along with other PAHs. The relative abundance of perylene from five-ring PAHs can be used to estimate the contribution of riverine-discharged PAHs to coastal sediments.

Polycyclic aromatic hydrocarbons in suspended particulate matter and sediments from the Pearl River Estuary and adjacent coastal areas, China.

The spatial distribution, composition, and sources of polycyclic aromatic hydrocarbons (PAHs) in sediments and suspended particulate matter (SPM) from the Pearl River Estuary and adjacent coastal areas were examined. Total PAH concentrations varied from 189 to 637 ng/g in sediments and 422 to 1,850 ng/g in SPM. PAHs were dominated by 5,6-ring compounds in sediments and by 2,3-ring compounds in SPM samples. Assessment of PAH sources suggested that biomass and coal combustion is the major PAH source to the outer part of the estuary sediments and that petroleum combustion is the major PAH source to the inner part of estuary sediments. As for SPM samples, PAH isomer pair ratios indicated multiple (petroleum, petroleum combustion, and biomass and coal combustion) PAH sources, and significant temporal variations could exist for the sources of water column PAHs in the study area. The distribution of perylene in SPM samples indicated that the river was the dominant source of perylene in SPM and that perylene could be taken as an index to assess the contribution of river inflow to the total PAHs in SPM samples. The high concentration of perylene in the sediment was indicative of an in situ biogenic origin.