ObjectiveTo detect the mutation of GJB2 gene in a Chinese family with Vohwinkel syndrome.MethodsClinical data were collected from 5 patients with Vohwinkel syndrome in a family,and blood samples were obtained from the 5 patients and 4 unaffected individuals in the family as well as from 100 normal human controls.Genomic DNA was extracted and subjected to PCR for the amplification of the entire encoding and flanking sequences of GJB2 gene(1015 bp) followed by bidirectional sequencing with the ABI PRISM 3730 automatic DNA sequencer.Finally,sequence alignment was carried out by using the software Sequencher 4.10.1 Demo.ResultsA heterozygous missense mutation 196G→C in the GJB2 gene,which resulted in the substitution of aspartic acid by histidine at codon 66 (D66H) in the first extracellular domain of the protein,was observed in all the patients of this family,but in none of the 4 unaffected individuals in this family or the 100 normal human controls.ConclusionThe D66H missense mutation in the GJB2 gene may contribute to the occurrence of Vohwinkel syndrome in Chinese Han population.

This study was aimed to analyze the formulas and classify the treatment of chronic kidney disease (CKD) by Dr. Huang Chunlin based on the data mining method. A total of 63 CKD outpatients' medical records by Dr. Huang were collected. And 226 formulas prescribed in the medical records were put in the Traditional Chinese Medicine Inheritance Support System (TCMISS). Data mining techniques were applied in the study of as-sociation rule mining algorithm and complex system entropy cluster, the usage frequency and the correlation of each single Chinese medicinal in order to sort out the prescription pattern of CKD. The results showed that to for-tify the spleen and tonify the kidney were found to be the basic method of CKD treatment . Through the applica-tion of TCMISS, the Chinese medicinals used in Dr. Huang's CKD treatment were Astragalus root, Eucommia, Epimedium, Cuscuta, Agastache, Danshen Root, Ligustrum. It was concluded that the TCMISS is of great value in the summarization of clinical experiences of well-known TCM doctors.

Purpose To analyze the perfusion differences of different pancreatic diseases using the low-dose whole organ dynamic volume CT perfusion imaging, and to provide the evidence for the clinical application. Materials and Methods Twenty-eight patients suspected as pancreatic disease were applied by 640 layer volume CT perfusion imaging for the pancreas. Data were collected at the same time of bolus injection of contrast agent, then were analyzed by spatial alignment and perfusion calculation using the perfusion software. The time-density curve, blood perfusion flow diagram and tissue artery blood flow were obtained using the maximum slope method. Results Normal pancreatic tissue (n=9) blood flow was (117.04±12.05) ml/(min?100 ml), pancreatitis organizations (6 cases with acute pancreatitis and 3 cases with chronic pancreatitis) (118.67±37.18) ml/(min?100 ml), pancreatic carcinoma tissue (n=10) was (67.16±18.94) ml/(min?100 ml). There was significant difference among three groups (F=8.59, P<0.001);the difference was demonstrated in pancreatic cancer vs. normal pancreas and pancreatic cancer vs. pancreatitis group (q=3.70, P<0.05), which could be clearly demonstrated by blood perfusion flow diagram. The difference was not statistically revealed pancreatitis and normal pancreas group (q=2.91, P>0.05). The total dose of X-rays in the whole scanning process was 21.5-23.9 mSv. Conclusion Low-dose whole pancreas organ CT perfusion scan can quantitatively analyze the hemodynamic changes in pancreatic disease, which be of great value for evaluating changes in microcirculation during the treatment of pancreatic cancer.

Objective To observe the surface area temperature of dysmenorrhea rats with cold stagnation syndrome; To compare the different effects of Sanyinjiao (SP6) and Guanyuan (RN4).Methods Forty female SD rats were randomized into control group, model group, Sanyinjiao (SP6) group, and Guanyuan (RN4) group, 12 rats in each group. Whole body freezing method combined with estradiol benzoate injection was used to establish models. Sanyinjiao (SP6) group and Guanyuan (RN4) group received moxibustion at corresponding points for 3 times after modeling. Infrared thermal imaging was used to measure the skin temperature at the surface projection area of Sanyinjiao (SP6) and Xuehai (SP10) and Sanyinjiao-Xuehai lines before and after moxibustion.Results Compared with control group, the temperature gap between double sides of Sanyinjiao-Xuehai lines significantly increased in model group 5-30 min after moxibustion (P<0.05). 30 min after moxibustion, the temperature of right Sanyinjiao significantly decreased in other three groups (P<0.01). Compared with model group, the temperature gap between double sides of Sanyinjiao-Xuehai lines significantly decreased after 5-30 min in Sanyinjiao group (P<0.05), while Sanyinjiao-Xuehai lines significantly decreased after 10-30 min in Guanyuan group (P<0.05). Comparison between two moxibustion groups, the influence of Sanyinjiao group to temperature gap between double sides of Sanyinjiao-Xuehai lines was earlier than Guanyuan group (P<0.05).Conclusion Moxbustion can decrease the temperature gap between double sides of Sanyinjiao-Xuehai lines and ease the imbalance. And the influence of moxbustion SP6 to temperature gap between double sides of Sanyinjiao-Xuehai lines is earlier than moxibustion RN4.

Human-like collagen (HLC) was cross-linked with hyaluronic acid by genipin in different ratio. The concentrations of hyaluronic acid in the mixture were 0, 0.01%, 0.05% and 0.1%. The blood vessel tubular grafts were then fabricated by freeze-drying. Microstructure, element composite, mechanical properties, cytotoxicity grade, and biocompatibility of different vascular scaffold groups were studied by scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), tensile test, burst pressure experiment, cytotoxicity experiment, endothelial cells planted in blood vessel scaffolds and hypodermic embedding of mice. The results showed that HLC-HA (0.05%) tubular scaffold exhibited interconnected well-distributed and porous structure and porosity of 94.38%; achieved the desirable mechanical property with stress of (1000.8 +/- 7.9) kPa and burst pressure of (1058.6 +/- 8.2) kPa, hypocytotoxicity, favourable cytocompatibility, hisocompatibility and disposition of degradation.
Adhesives ; chemistry ; Animals ; Biocompatible Materials ; chemistry ; Biomimetic Materials ; chemistry ; Blood Vessels ; drug effects ; physiology ; Collagen ; chemistry ; Humans ; Hyaluronic Acid ; chemistry ; Iridoid Glycosides ; Iridoids ; chemistry ; Materials Testing ; Mice ; Tissue Engineering ; instrumentation ; methods ; Tissue Scaffolds

Objective:To investigate the effect of low-frequency ultrasound combined with hydroxychloroquine (HCQ) on the growth and invasion of 4T1 breast cancer in mice.Methods:4T1 cells in logarithmic growth phase were divided into 4 groups: control group, ultrasound group, HCQ group and ultrasound combined with HCQ group. Western blot was performed to detect the effects of ultrasound combined with HCQ on the protein expression levels of autophagy-related proteins LC3, p62 and matrix metalloproteinase 9 (MMP-9). Transmission electron microscopy was used to observe the formation of autophagic vesicles. The cell proliferation and cell viability were determined by EdU and CCK-8. Transwell assay was performed to detect the effect of ultrasound combined with HCQ on the invasive ability of 4T1 cells. Flow cytometry was performed to detect the effect of ultrasound combined with HCQ on the apoptosis of 4T1 cells. The transplantation tumor model of 4T1 breast cancer in BALB/c mice was constructed.The mice were randomly divided into 4 groups ( n=5 for each group), including control group, ultrasound group, HCQ group, ultrasound combined with HCQ group. The tumor volume and mice body weight were evaluated and measured in each group every 2 days. Results:The expression of LC3-Ⅱ and p62 protein levels increased in the ultrasound combined with HCQ group, and intracellular autophagosome accumulation was evident by transmission electron microscopy. In cellular experiments, compared with the other groups, the ultrasound combined with HCQ group showed stronger growth inhibition, significantly decreased cell proliferation rate, decreased expression of MMP-9, and significantly inhibited invasion (all P<0.050). In the in vivo experiments, compared with the control group, the tumor growth rate of all 3 inter vention groups of mice decreased (all P<0.050), and the ultrasound combined with HCQ group had better therapeutic effects than the ultrasound and HCQ groups. The treatment effect of ultrasound combined with HCQ group was better than that of ultrasound and HCQ groups (all P<0.001). Conclusions:The combination of low-frequency ultrasound and hydroxychloroquine could synergistically inhibit tumor cell proliferation, promote apoptosis, and significantly inhibit tumor growth in 4T1 tumor-bearing mice.

Objective To observe the analgesic effect of acupuncture at Guanyuan (CV4) and its effect on vasomotor substances in rats with dysmenorrhea due to coagulated cold syndrome. Method The coagulated-cold dysmenorrhea rat model was developed by Estrodiol benzoate and Oxytocin injectin plus physical freezing. The writhing response (writhing latency, writhing frequency, and writhing score) was observed, and the contents of TXB2 and 6-keto-PGF1a were detected by using enzyme-linked immunosorbent assay (ELISA). Result Compared with the saline water group, the writhing latency was significantly shortened, the writhing frequency was significantly increased, and the writhing score was more significantly increased in the model group (P<0.01);compared with the model group, the writhing latency was significantly prolonged, the writhing frequency was decreased, and the writhing score was significantly lower in the acupuncture group (P<0.05, P<0.01). Compared with saline water group, the content of plasma 6-keto-PGF1a was significantly lower (P<0.05) and the content of plasma TXB2 showed an increasing tendency (P>0.05) in the model group. Compared with the model group, the content of plasma 6-keto-PGF1a showed an increasing tendency (P>0.05) and the content of plasma TXB2 showed a decreasing tendency (P>0.05) in the acupuncture group. Conclusion The vasomotor substances are obviously disordered in the blood of cold-coagulated dysmenorrhea rat models. Acupuncture at Guanyuan can improve the writhing response and release pain, and meanwhile positively regulate the vasomotor substances such as TXB2 and 6-keto-PGF1a. The vasomotor substances are plausibly one of the major substances in the action of acupuncture in preventing and treating dysmenorrhea.

Objective To explore the effect of different acupuncture stimuli on uterine micro-circulation of dysmenorrheal rats with cold stagnation syndrome. Methods Totally 32 three-month old female SD rats in diestrus were randomly divided into saline control group, model group, A stimuli group, and B stimuli group, 8 rats in each group. Model group and treatment groups were given whole body freezing combined with estradiol benzoate injection method to establish models. A stimuli group was given deep puncture with manipulation, while B stimuli group was treated by shallow puncture without manipulation. Diameter of uterine capillary,micro-vessel, TXB2, and 6-keto-PGF1αlevels were observed in each group. Results Compared with the saline group, capillary diameter in model group was significantly reduced at 5, 10, 20, 30 min time point (P<0.01);micro-vascular diameter was significantly reduced at 5, 10, 20, 30, 40 min time point (P<0.01);plasma 6-keto-PGF1α levels decreased (P<0.01);TXB2/6-keto-PGF1αincreased significantly (P<0.01). Compared with model group, capillary diameter in A stimuli group enlarged at 5, 10, 20, 30 min time point (P<0.05), micro-vascular diameter dilated at 5, 10, 20, 30, 40 min time point (P <0.01), plasma 6-keto-PGF1α level increased (P <0.05), TXB2/6-keto-PGF1αdecreased significantly (P<0.05);micro-vascular diameter in B stimuli group dilated at 20, 30 min time point (P<0.05), plasma TXB2/6-keto-PGF1α decreased significantly (P<0.05). Compared with B stimuli group, capillary diameter in A stimuli group dilated at 5, 10, 20, 30 min time point (P<0.05) and micro-vascular diameter dilated at 5, 10, 20, 30, 40 min time point significantly (P<0.01). Conclusion Dysmenorrheal rats with cold stagnation syndrome show obvious disorder of the uterus micro-circulation and circulation related substances. Both A and B acupuncture stimuli improved uterus micro-circulation of dysmenorrheal rats with cold stagnation syndrome, and its mechanism may be related to the recovery the balance between TXB2 and 6-keto-PGF1α.

Objective Infrared thermal imaging can be applied to the diagnosis and auxiliary diagnosis of some diseases . The aim of this study is to explore acupuncture-induced changes in skin temperature in acupoint areas and whether skin temperature in -creases or decreases in the acupoint areas along meridians . Methods Thirty two female SD rats were randomly and equally divided into four groups:saline control,cold congealing and dysmenorrhea model , Sanyinjiao (SP6), and Guanyuan( CV4).Models were es-tablished in the latter three groups by subcutaneous injection of estradiol benzoate at 0.5 mg for 10 successive days and , 1hour after the last administration , intraperitoneal injection of oxytocin at 2 U, followed by exposure of the rats to-25℃in a freezer 4 hours a day for 5 days.Meanwhile , the control rats received normal saline only and were not exposed to low temperature .Infrared thermal imaging was used to measure the skin temperature at the acupoint areas of SP6, Xuehai (SP10), and CV4 before and at 0, 5, 10, 20, 30, 40, 50 and 60 min after needling . R esults At 0 to 5 min after nee-dling, the skin temperature of the left SP6 and right SP10 was signifi-cantly decreased in both the SP6 and CV4 groups ( [ -0. 56 ± 0.22]℃and [-0.48 ±0.11]℃, P<0.01), and so was that of the right SP10 ([ -0.64 ±0.21]℃ and [ -0.45 ±0.13]℃, P<0.05).At 5 to 10min, the skin temperature of the right SP6 and SP10 was markedly increased in the SP6 group ([-0.49 ±0.35]℃and [-0 .18 ±0.20]℃, P<0.01), and so was that of the right SP6 in the SP6 group at 20 to 30 min ([ -0.14 ±0.25]℃) as compared with the model and CV4 groups (P<0.01).At 30 to 40 min, the skin temperature of the right SP10 was remarkably elevat-ed in the SP6 group ([ -0.03 ±0.11]℃) in comparison with the model group (P<0.01).No significant differences were observed in the skin temperature of the left SP10 and CV4 at different time points among the four groups (P>0.05). Conclusion The skin temperature of SP6 and SP10 can be regulated by needling both the acupoints of SP 6 and CV4.The increase in the skin temperature of the right SP6 and SP10 in the SP6 group and no change in the CV 4 group indicated dynamic temperature changes in the acupoint area along the meridian after needling.

A case of ichthyosis follicularis,alopecia and photophobia syndrome caused by a novel mutation c.1165C＞T in the membrane-bound transcription factor protease site 2 (MBTPS2) gene was firstly reported.The proband presented with dry skin,congenital hairlessness,follicular keratotic papules,photophobia,epilepsy,and mental and motor retardation.Next-generation and Sanger sequencing analysis confirmed that the proband and his mother both had a c.1165C＞T (p.pro389Ser) mutation in exon 9 of the MBTPS2 gene.According to the clinical manifestations of the patient and genetic characteristics of the MBTPS2 gene mutation,the patient was diagnosed with ichthyosis follicularis,alopecia and photophobia syndrome.