Transdermal Delivery of Recombinant Human Growth Hormone by Liposomal Gel for Skin Photoaging Therapy.

Human growth hormone (hGH) has emerged as a promising therapeutic agent to prevent and treat skin photoaging. However, the success of hGH therapy largely lies in the availability of an optimal delivery system that enables the efficient delivery of hGH to the dermal layer of the skin. Here, we report a delivery system of hyaluronic acid/liposome-gel-encapsulated hGH (HA/HL-Gel) that can transdermally deliver hGH into the skin for hGH-based photoaging therapy through the upregulation of collagen type I (collagen-I). Specifically, hGH-liposomes were prepared by ethanol injection and then modified with HA to achieve specific targeting. The best formulation of HA/hGH-liposomes (HA/HL) had a high encapsulation efficiency (about 20%), with a size of 180 ± 1.2 nm. The optimized HA/HL was further incorporated into the carbomer gel to form an HA/HL-Gel. The biological activity of HA/HL on human dermal fibroblasts (HDFs) was confirmed by the elevated expression level of collagen-I through the enhanced local formation of insulin-like growth factor-1 (IGF-1) in the photoaging model. Moreover, HA/HL-Gel reduced ultraviolet (UV)-induced erythema and wrinkle formation. Meanwhile, immunohistochemical staining further showed higher levels of collagen-I in the HA/HL-Gel group compared to other groups tested. Taken together, these results demonstrate that HA/HL-Gel treatment could significantly ameliorate skin photoaging and thus may be used as a clinical potential for antiaging therapy.

Exosomes in the tumor microenvironment: Promoting cancer progression.

Exosomes, which are extracellular vesicles produced by endosomes, are important performers of intercellular communication functions. For more than three decades, there has been a growing awareness of exosomes as the contents of the tumor microenvironment and their intimate connection to the development of cancer. The composition, generation, and uptake of exosomes as well as their roles in tumor metastasis, angiogenesis, and immunosuppression are discussed in this paper. In order to stop the progression of cancer, it is crucial to find new diagnostic biomarkers and therapeutic targets for the disease. Knowing the biological characteristics of exosomes and their functions in tumor development helps in this endeavor.

Assessment of peripheral neuropathy in type 2 diabetes by diffusion tensor imaging.

BACKGROUND: To evaluate the diagnostic accuracy of diffusion tensor imaging (DTI) in diabetic peripheral neuropathy (DPN) for patients with type 2 diabetes and detect the correlations with electrophysiology. METHODS: A total of 27 patients with type 2 diabetes with DPN, 24 patients with type 2 diabetes without peripheral neuropathy (NDPN), as well as 32 healthy controls (HC) were enrolled in this study. Clinical examinations and neurophysiologic tests were used to determine the presence of DPN. Fractional anisotropy (FA) and apparent diffusion coefficient (ADC) of peripheral nerves, including the tibial nerve (TN) and common peroneal nerve (CPN), were calculated. Receiver operating characteristic (ROC) analysis was performed for FA and ADC values. Pearson's correlation coefficient was used to assess the correlation between DTI and electrophysiology parameters in the patient group. RESULTS: The tibial and common peroneal nerve FAs were lowest (P=0.003, 0.001, respectively) and ADC was highest (P=0.004, 0.005, respectively) in the DPN group. The FA value of the axonal injury group was lower than that in the demyelination group (P=0.035, 0.01, respectively), while the ADC value was higher (P=0.02, 0.01, respectively). In the DPN group, FA value was positively correlated with motor conduction velocity (MCV) (tibial nerve: r=0.420, P=0.007; common peroneal nerve: r=0.581, P<0.001) and motor amplitude (MA) (tibial nerve: r=0.623, P<0.001; common peroneal nerve: r=0.513; P=0.001), while ADC values was negatively correlated with MCV (tibial nerve: r=-0.320, P=0.044; common peroneal nerve: r=-0.569; P<0.001), and MA (tibial nerve: r=-0.491, P=0.001; common peroneal nerve: r=-0.524; P=0.001). CONCLUSIONS: With a lower FA value and higher ADC value, DTI accurately discriminated DPN. The DTI multi-parameter quantitative analysis of peripheral nerves differentiated DPN axonal injury from the demyelinating lesion, and hence, could be applied in the diagnosis of DPN.

Exosomes as smart drug delivery vehicles for cancer immunotherapy.

Exosomes (Exos) as drug delivery vehicles have been widely used for cancer immunotherapy owing to their good biocompatibility, low toxicity, and low immunogenicity. Some Exos-based cancer immunotherapy strategies such as tuning of immunosuppressive tumor microenvironment, immune checkpoint blockades, and cancer vaccines have also been investigated in recent years, which all showed excellent therapeutic effects for malignant tumor. Furthermore, some Exos-based drug delivery systems (DDSs) for cancer immunotherapy have also undergone clinic trails, indicating that Exos are a promising drug delivery carrier. In this review, in order to promote the development of Exos-based DDSs in cancer immunotherapy, the biogenesis and composition of Exos, and Exos as drug delivery vehicles for cancer immunotherapy are summarized. Meanwhile, their clinical translation and challenges are also discussed. We hope this review will provide a good guidance for Exos as drug delivery vehicles for cancer immunotherapy.

Clinical value of high-resolution dynamic contrast-enhanced (DCE) MRI in diagnosis of cutaneous squamous cell carcinoma.

BACKGROUND: While uncomplicated cases of skin squamous cell carcinoma (cSCC) can be treated with surgery topical therapy alone, more objective and non-invasive examination methods are needed to guide clinicians to make more detailed biopsy and surgical plans for lesions with atypical or subcutaneous growth. High-resolution magnetic resonance imaging (HR-MRI) is a novel skin imaging method. MATERIALS AND METHODS: Prospective collection of 19 patients with clinically suspected cSCC. All patients underwent high-resolution DCE-MRI using a 70-mm microscopy coil before operation. The imaging features and results of surgical pathology were recorded. Ktrans , Kep , Ve values, and the time-signal curve (TIC) types were determined using DCE images. RESULTS: 16 cases of cSCC, 3 cases of acanthoma. The subcutaneous invasion of all lesions was clearly displayed, of which 8 lesions invaded the subcutaneous fat layer, 5 invaded the muscle layer, 1 invaded the periosteum, 2 invaded the cap fascia, and the layer of all lesions invasion judged by HR-MR imaging was consistent with the postoperative pathology. The main manifestations of cSCC were ill-defined margin, obvious inhomogeneous enhancement, higher perfusion parameters value and type-III TIC, while acanthoma showed well-defined and type-I TIC. Some imaging findings (such as boundary, enhancement) and DCE perfusion parameters of the two groups overlap. CONCLUSION: High-resolution DCE-MRI can fully and directly display the subcutaneous invasion of cSCC, and more work needs to be done to prove its value. Next, we will expand the sample size, and further explore its value in the differential diagnosis and prognosis evaluation of cSCC from acanthoma or other skin tumors.

Bone marrow adipose amount influences vertebral bone strength.

Association of bone marrow adipose and microstructure with bone strength in osteoporotic rats using MR Dixon analysis and micro-CT was evaluated. A total of 40 female Sprague-Dawley rats (6-month-old) were divided randomly into sham-operated (SHAM, n=20) group and ovariectomized (OVX, n=20) group. Fat fraction (FF) was measured by two-point Dixon method with MR imaging at the baseline, 4th, 8th and 12th week, respectively. After sacrifice by anesthesia, the fifth lumbar vertebrae bone was sampled for micro-CT scanning. The biomechanical analysis was also performed. FF in osteoporotic rats significantly increases with time, which correlates with bone microstructure parameters. Compared with biomechanical test, FF showed negative correlation with break stress and elastic modulus. It also suggested that loss of bone mass was accompanied with the increase of adipose tissue content in vertebrae bone marrow. The impairment of bone strength leads to the risk of brittle fracture. In conclusion, the bone marrow adipose amount obtained by MR Dixon and microstructure by micro-CT correlates to bone strength in osteoporotic rats.

Atypical imaging observations of branchial cleft cysts.

The aim of the present study was to assess the atypical imaging manifestations of branchial cleft cysts (BCCs) confirmed by pathology. Computerized tomography (CT) or magnetic resonance imaging (MRI) of 17 BCC cases were reviewed. The imaging features, including laterality, location, border, attenuation and internal architecture, were evaluated. All 17 cases were second BCCs, including 5 cases of Bailey type I classification cysts and 12 cases of type II classification cysts. The atypical imaging features included signal and morphological abnormalities. The abnormal signal intensities were caused by intracapsular bleeding (n=2) or solidification of cystic fluid (n=2). Intracystic hemorrhaging revealed homogeneous hyperintensity on T1-weighted image (T1WI) and T2-weighted image (T2WI). Solidification of cystic fluid revealed slightly homogeneous hyperintensity compared with muscle on T1WI and homogeneous hypointensity on T2WI without enhancement. The aberrant morphology mainly presented as thickening of the cystic wall (n=13). Thickened walls of BCCs with ill- (n=5) or well- (n=8) defined borders were observed in 13 patients. In 3 patients, significant enhancement was identified following intravenous gadolinium administration (n=4). When with atypical CT or MRI features are presented, the typical location of BCCs can help in the diagnosis, as it is located at the lateral portion of the neck adjacent to the anterior border of the mandibular angle or sternocleidomastoid muscle. The atypical observations, including variable signals, imply that the cystic content has changed. Thickened walls indicate inflammation or cancerous tendency and patients with ill-defined margins, vascular involvement or lymphadenopathy atelectasis indicate malignant conversion.

[Expertise-based randomized controlled trials and its application in acupuncture clinical research].

Through introduction of the methodological mechanism and comparison with classic randomized controlled trial, the status and the applicability of the expertise-based randomized controlled trials in clinic are explored, and its characteristics in acupuncture clinical application are analyzed. It is held that expertise-based randomized controlled trial is more suitable for the acupuncture clinical research, especially for acupuncture practice which emphasizes manipulations and different schools.

Rapid detection of intestinal pathogens in fecal samples by an improved reverse dot blot method.

AIM: To develop a new, rapid and accurate reverse dot blot (RDB) method for the detection of intestinal pathogens in fecal samples. METHODS: The 12 intestinal pathogens tested were Salmonella spp., Brucella spp., Escherichia coli O157:H7, Clostridium botulinum, Bacillus cereus, Clostridium perfringens, Vibrio parahaemolyticus, Shigella spp., Yersinia enterocolitica, Vibrio cholerae, Listeria monocytogenes and Staphylococcus aureus. The two universal primers were designed to amplify two variable regions of bacterial 16S and 23S rDNA genes from all of the 12 bacterial species tested. Five hundred and forty fecal samples from the diarrhea patients were detected using the improved RDB assay. RESULTS: The methods could identify the 12 intestinal pathogens specifically, and the detection limit was as low as 103 CFUs. The consistent detection rate of the improved RDB assay compared with the traditional culture method was up to 88.75%. CONCLUSION: The hybridization results indicated that the improved RDB assay developed was a reliable method for the detection of intestinal pathogen in fecal samples.

[Detection and identification of seven clinical common pathogenic bacteria by oligonucleotide microarray].

OBJECTIVE: Using 16S rDNA and 23S rDNA genes as the target sequences to develop a system based on oligonucleotide microarray and to detect the seven clinical pathogenic bacteria, commonly seen. METHODS: Double polymerase chain reaction (PCR) was applied to amplify the segments of 16S rDNA and 23S rDNA genes of the target bacteria. An oligonucleotide microarray was constructed to simultaneously detect EHEC O157:H7, Vibrio parahaemolyticus, Salmonella sp., Vibrio cholerae, Listeria monocytogenes, Campylobacter jejuni and Shigella sp. Specificity, sensitivity and reproducibility of the microarray during detection were checked. And then microarray was used to detect the microbes in stool specimens of 81 patients with diarrhea and vomiting. RESULTS: The double PCR method could simultaneously amplify the target sequences of 16S rDNA and 23S rDNA genes of the seven pathogens. The sensitivity of the developed oligonucleotide microarray could reach 10(3) cfu/ml but no positive results were presented for non-targeted bacteria. The coefficients of differentiation in one lot or among different lots of the microarray slices were 3.89% - 5.81%. The positive detection rate of the stool specimens by oligonucleotide microarray was 39.5% (32/81), with a coincidence of 96.3% (78/81) for the patients and another coincidence of 96.8% (31/32) for bacterial genus or species identification, when comparing to the results by routine bacteriological examinations. CONCLUSION: The established assay in this study based on oligonucleotide microarray to detect the seven pathogenic bacteria has many advantages such as convenient, rapid, accurate, stable and high flux, which is suitable for clinical specimen examination and epidemiological field investigation.

[The development and clinical application of papillomavirus genotyping by HPV DNA suspension array the development and clinical application of papillomavirus genotyping by HPV DNA suspension array].

OBJECTIVE: To develop a new platform for genotyping human papillomavirus(HPV) and to investigate its effect in clinical application. METHODS: By combining L1 consensus PCR and multiplex hybridization using a Luminex xMAP system-based suspension array, we developed a rapid high-throughput assay,the HPV DNA suspension array (HPV-SA), capable of simultaneously typing 30 HPVs, including 18 high-risk HPV genotypes and 12 low-risk HPV genotypes. 810 clinical specimens were used to investigate the effect of HPV-SA. Veracity of the genotyping result was verified by E7 type-specific PCR-DNA sequencing. RESULTS: Among the 810 clinical specimens, 243 were found to be HPV positive,including high-risk HPV subtypes 16, 18, 26, 31, 33, 39, 45, 51, 53, 56, 58, 59, 66, 68, 73 and 82,and low-risk HPV6, 11, 34, 54, 61, 67, 70 and 84. The sensitivity tested by standard samples was up to 10 copies of HPV DNA. CONCLUSION: The HPV-SA developed here showed high sensitivity and specificity, suitable to be applied in clinical practice for HPV diagnosis and investigation on the prevalence of HPV sub-types.

[Detection and identification of human metapneumovirus by real time reverse transcription PCR].

OBJECTIVE: To develop a rapid, sensitive and specific real time reverse transcription PCR for detecting and identifying human metapneumovirus. METHODS: The Hmpv-L gene of human metapneumovirus was chosen as target gene, the primers and TaqMan probe were designed, and the PCR reaction was optimized systematically. The total RNA was extracted from respiratory specimens, and reverse transcription was performed through random primer. The cDNA was detected by using real time PCR. The specificity, sensitivity and reproducibility of real time PCR were estimated. The real time PCR was applied to detect 180 clinical respiratory specimens. RESULTS: The human metapneumovirus can be detected using real time reverse transcription PCR accurately and quickly, and the sensitivity was 1 copy/microl. The coefficient of variation of intra-assay and inter-assay was less than 5%. Among those 180 specimens, 28 (15.56%) were positive for human metapneumovirus, the clinical diagnoses for these 28 patients were pneumonia (15.60%, 17/109) and bronchiolitis (15.49%, 11/71). 21 positive specimens were from patients under 2 years of age, and 6 positive specimens were from patients between 2 and 5 years of age, only 1 positive specimens was from patients over 5 years. CONCLUSION: It is demonstrated that real time reverse transcription PCR is a reliable, accurate and feasible assay for human metapneumovirus, which has become one of the most important pathogens induced acute respiratory infections in pediatric patients.