Expert consensus on late stage of critical care management / 中华内科杂志

We wished to establish an expert consensus on late stage of critical care (CC) management. The panel comprised 13 experts in CC medicine. Each statement was assessed based on the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) principle. Then, the Delphi method was adopted by 17 experts to reassess the following 28 statements. (1) ESCAPE has evolved from a strategy of delirium management to a strategy of late stage of CC management. (2) The new version of ESCAPE is a strategy for optimizing treatment and comprehensive care of critically ill patients (CIPs) after the rescue period, including early mobilization, early rehabilitation, nutritional support, sleep management, mental assessment, cognitive-function training, emotional support, and optimizing sedation and analgesia. (3) Disease assessment to determine the starting point of early mobilization, early rehabilitation, and early enteral nutrition. (4) Early mobilization has synergistic effects upon the recovery of organ function. (5) Early functional exercise and rehabilitation are important means to promote CIP recovery, and gives them a sense of future prospects. (6) Timely start of enteral nutrition is conducive to early mobilization and early rehabilitation. (7) The spontaneous breathing test should be started as soon as possible, and a weaning plan should be selected step-by-step. (8) The waking process of CIPs should be realized in a planned and purposeful way. (9) Establishment of a sleep-wake rhythm is the key to sleep management in post-CC management. (10) The spontaneous awakening trial, spontaneous breathing trial, and sleep management should be carried out together. (11) The depth of sedation should be adjusted dynamically in the late stage of CC period. (12) Standardized sedation assessment is the premise of rational sedation. (13) Appropriate sedative drugs should be selected according to the objectives of sedation and drug characteristics. (14) A goal-directed minimization strategy for sedation should be implemented. (15) The principle of analgesia must be mastered first. (16) Subjective assessment is preferred for analgesia assessment. (17) Opioid-based analgesic strategies should be selected step-by-step according to the characteristics of different drugs. (18) There must be rational use of non-opioid analgesics and non-drug-based analgesic measures. (19) Pay attention to evaluation of the psychological status of CIPs. (20) Cognitive function in CIPs cannot be ignored. (21) Delirium management should be based on non-drug-based measures and rational use of drugs. (22) Reset treatment can be considered for severe delirium. (23) Psychological assessment should be conducted as early as possible to screen-out high-risk groups with post-traumatic stress disorder. (24) Emotional support, flexible visiting, and environment management are important components of humanistic management in the intensive care unit (ICU). (25) Emotional support from medical teams and families should be promoted through"ICU diaries"and other forms. (26) Environmental management should be carried out by enriching environmental content, limiting environmental interference, and optimizing the environmental atmosphere. (27) Reasonable promotion of flexible visitation should be done on the basis of prevention of nosocomial infection. (28) ESCAPE is an excellent project for late stage of CC management.

Characterization and genomic sequencing of PS2, a lytic phage that infects Serratia marcescens clinical isolates / 中国人兽共患病学报

The present study focused on the characterization and genomic sequence of phage PS2 that infects Serratia marcescens clinical isolates.The morphology of phage PS2 was observed with electron microscope.The one-step growth curve,host range,and stability of PS2 were investigated.In addition,Phage DNA was extracted from the purified phage particles using a MiniBEST Viral RNA/DNA Extraction Kit.DNA sample was analyzed by digesting with restriction enzymes.The phage DNA was used for constructing the sequencing library.The library was sequenced on a MiSeqTM platform.The whole genome sequence was obtained by Velvet (version:1.2.08) assembling.Phage PS2 belongs to the Myoviridae family.The linear,circularly permuted,167 266-bp double-stranded DNA genome of PS2 has high similarities to T4-1ike phages.The phage DNA contains 41.7％ GC and 276 ORFs.PS2 exhibited a 21-minute latent period and 70 PFU per cell at burst size when the pathogenic S.marcescens strain S2 served as a host.Further investigation suggested that PS2 is stable in a wide pH range (pH5 to pH10) and at extreme temperatures (50 ℃ and 60 ℃) after incubation alone at different pHs and different temperatures,respectively.The paper focused on the isolation and identification of a novel lytic S.marcescens phage,the biological characteristics,the whole genome sequencing and the preliminary study of bioinformatics,which laid the foundation for deeply analysis to the phage therapy of multi-drug resistant bacteria and the phage biological information.

Characterization and genomic sequencing of PS2, a lytic phage that infects Serratia marcescens clinical isolates / 中国人兽共患病学报

The present study focused on the characterization and genomic sequence of phage PS2 that infects Serratia marcescens clinical isolates.The morphology of phage PS2 was observed with electron microscope.The one-step growth curve,host range,and stability of PS2 were investigated.In addition,Phage DNA was extracted from the purified phage particles using a MiniBEST Viral RNA/DNA Extraction Kit.DNA sample was analyzed by digesting with restriction enzymes.The phage DNA was used for constructing the sequencing library.The library was sequenced on a MiSeqTM platform.The whole genome sequence was obtained by Velvet (version:1.2.08) assembling.Phage PS2 belongs to the Myoviridae family.The linear,circularly permuted,167 266-bp double-stranded DNA genome of PS2 has high similarities to T4-1ike phages.The phage DNA contains 41.7％ GC and 276 ORFs.PS2 exhibited a 21-minute latent period and 70 PFU per cell at burst size when the pathogenic S.marcescens strain S2 served as a host.Further investigation suggested that PS2 is stable in a wide pH range (pH5 to pH10) and at extreme temperatures (50 ℃ and 60 ℃) after incubation alone at different pHs and different temperatures,respectively.The paper focused on the isolation and identification of a novel lytic S.marcescens phage,the biological characteristics,the whole genome sequencing and the preliminary study of bioinformatics,which laid the foundation for deeply analysis to the phage therapy of multi-drug resistant bacteria and the phage biological information.

Simultaneous determination of bisphenols and alkylphenols in water by solid phase extraction and ultra performance liquid chromatography-tandem mass spectrometry / 生物医学与环境科学（英文）

To establish an analytical method for determination of four bisphenols (BPA, BPB, BPF, and BPS) and two alkylphenols (4-n-OP, 4-n-NP) in water by ultra performance liquid chromatography- tandem mass spectrometry (UPLC/MS/MS). The water samples were extracted and condensed with solid-phase extraction (SPE) using C18 cartridges and eluted by acetonitrile. Separation was carried out with Acquity BEH C8 column and detection were performed by UPLC/MS/MS. Quantification was calculated by using the internal standard BPA-d16 and 4-n-NP-d8. The linear correlation coefficients of these compounds in the range of 1.0-100.0 μg/L were all over 0.999. The minimum detectable concentrations were 0.75-1.0 ng/L, and the recoveries ranged from 87.0% to 106.9%. Relative standard deviations (RSDs) were between 1.26% and 3.67%. Applying this method to detect the source water of Chaohu Lake and drinking water of Hefei, six target compounds were detected in different levels. This method is simple with high sensitivity and selectivity, could be suitable for the determination of these compounds in source and drinking water.

Application of surface enhanced laser desorption/ionization time-of-flight mass spectrometry in the diagnosis of leukemia / 中国实验血液学杂志

This study was purposed to find new biomarkers and to establish protein finger print model for diagnosis of leukemia. A total of 40 leukemia samples and 37 healthy control samptes were tested by surface enhance laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF- MS). The data of spectra were analyzed by bioinformatics tools like Biomarker Patterns 5.0 and discriminant analysis to establish diagnostic mode1. The results showed that 22 protein features were stably detected by protein fingerprint, The detective model combined with 3 biomarkers (m/z 4650, 8609 and 11660) could differentiate leukemia with sensitivity of 97.5% (39/40) and specificity of 91.9%(34/37). It is concluded that the detective model established by 3 protein features may be a novel method for diagnosis of leukemia.

Clinical research of medicinal vesiculation for perennial allergic rhinitis / 中国针灸

<p><b>OBJECTIVE</b>To compare the efficacy differences between dog-days medicinal vesiculation and regular-day medicinal vesiculation for perennial allergic rhinitis (PAR), and observe their effects on serum immune globulin E (IgE) and interleukin-4 (IL-4).</p><p><b>METHODS</b>Seventy-two patients were randomly divided into a dog-days moxibustion group (34 cases) and a regular-day moxibustion group (38 cases). In the dog-days moxibustion group, medicinal vesiculation was applied on the 1st dog-day, 2nd dog-day and last dog-day in summer by lunar calendar, 3 treatments per dog-day for totally 9 times. In the regular-day moxibustion group, the moxibustion was given on the regular day for continuous 9 times. The symptom score, rhinoconjunctivitis quality of life questionnaire (RQLQ) and the level of IgE and IL-4 were compared before and after treatment in two groups; the short-term and two-year efficacy evaluation were performed too.</p><p><b>RESULTS</b>The short-term total effective rate was 88.2% (30/34) in the dog-days moxibustion group, which was not significantly different to 86.8% (33/38) in the regular-day moxibustion group (P>0.05). The long-term total effective rate was 97.1% (33/34) in the dog-days moxibustion group, which was significantly superior to 81.6% (31/38) in the regular-day moxibustion group (P<0.05). After treatment, the serum IgE, IL-4 and RQLQ were significantly reduced (all P<0.01), but the difference between two groups was not significant (all P>0.05).</p><p><b>CONCLUSION</b>Medicinal moxibustion could be taken as a regular treatment for PAR, which could be performed during the whole year, and dog-days moxibustion could be considered as an enhanced method for prevention and treatment of PAR.</p>

Up-Regulation of Cyclooxygenase 2 and Matrix Metalloproteinases-2 and -9 in Cutaneous Squamous Cell Carcinoma: Active Role of Inflammation and Tissue Remodeling in Carcinogenesis

BACKGROUND: Tissue inflammation and remodeling have been extensively studied in various tumors in relation with their invasiveness and metastasis. OBJECTIVE: The purpose of this study was to investigate the change in tissue inflammation and remodeling markers in cutaneous squamous cell carcinoma (SCC). METHODS: Expression levels of cyclooxygenase-2 (COX-2) as an inflammatory marker and matrix metalloproteinases-2 and -9 (MMPs 2/9) as remodeling markers were studied in mouse and human SCCs. Western blot analysis and RT-PCR for COX-2 and MMPs 2/9 were performed with skin samples from SCC patients and chronic ultraviolet B (UVB)-induced SCC from hairless mice. RESULTS: mRNA and protein levels of COX-2 and MMPs 2/9 were up-regulated with the higher sensitivity for MMP-9 in mouse SCCs, which were induced by chronic UVB irradiation. Consistently, COX-2 and MMPs 2/9 were up-regulated with the higher sensitivity for MMP-9 in human SCCs. CONCLUSION: COX-2 and MMPs 2/9 are up-regulated in well-differentiated cutanous SCC. Our findings indicate that inflammatory and tissue remodeling processes are actively induced during carcinogenesis of cutaneous SCC.

Evaluation of the research features and methodology of acupuncture literatures of headache published in foreign journals in SCI in the recent 5 years / 中国针灸

Retrieving literatures in SCI source journals in the recent 5 years through Web of Science and PubMed databases, articles regarding to the treatment of headache with acupuncture are arranged and analyzed. It turns out that (1) the acupoint selection and treatment is not under the guidence of syndrome differentiation in the clinical research literatures published in foreign journals. (2) The clinical research trials published in foreign journals shows that regarding to curative effect, there is no difference between acupuncture group and sham acupuncture group which indicats that the westerners do not grasp the westerners do not grasp the essene of acupuncture techniques owing to the great difference of medical theory and thinking mode between eastern and western, and the western's shallow cognition of TCM and acupuncture. The sham acupuncture trials published in foreign journals with respect to acupuncture for headache cannot highlight the advantage o acupuncture therapy, what's more, the sham acupuncture trial designation is unfit for acupuncture therapy.

Spermatic cord liposarcoma: a case report and review of the literature / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the clinical features and treatment of spermatic cord liposarcoma (SCL).</p><p><b>METHODS</b>We retrospectively analyzed the clinical data of a case of SCL, reviewed the related literature and investigated the diagnosis and treatment of the disease.</p><p><b>RESULTS</b>The patient underwent tumor resection and left inguinal orchidectomy. Postoperative pathology confirmed the case to be s SCL. Neither recurrence nor metastasis was found during the five-month follow-up.</p><p><b>CONCLUSION</b>SCL is a rare medical condition with no specific imaging and laboratory features. Radical orchidectomy with wide local excision of the mass is recommended for its treatment, and adjuvant radiotherapy can be considered in intermediately or highly differentiated tumors and recurrent liposarcomas, while the role of chemotherapy is not well-defined.</p>

Correlation analysis between hemostatic activity and commercial grades of Panax notoginseng / 中草药

Objective: To explore the correlation between the hemostatic activity and commercial grades of Panax notoginseng. Methods: After treatment with different commercial grades of P. notoginseng, the mice blooding time (BT) and clotting time (CT) were measured. At the same time the prothrombin time (PT), activation part thrombin time (APTT), fibrinogen (FIB), and platelet count (PLT) were also detected by fully automated blood cell analyzer. The contents of dencichine were determined by HPLC. Results: Mice BT, CT, PT, and APTT were decreased and PLT was increased significantly in all the commercial grades of P. notoginseng. Among all the commercial grades, the countless Tou Sanqi Powder has the best hemostatic activity. There is no correlation between the commercial grades and homostatic activity. Different contents of the extracted dencichine were observed among different commercial grades of P. notoginseng. The highest content of dencichine was 0.98% in countless Tou Sanqi and the lowest was 0.52% in 80 Tou Sanqi. There is no statistical correlation between the dencichine and hemostatic activity or dencichine and commercial grades of P. notoginseng. Conclusion: There is no statistical correlation between the commercial grades of P. notoginseng and haemostatic activity or dencichine contents. The dencichine in countless Tou Sanqi which is the cheapest of all displays the better hemostaic activity than those in 13 or 20 Tou Sanqi whose cost is almost as ten times as countless Tou Sanqi's.

The effects of TiO_2 nano thin film on the migration and adherence of BLEC in vitro / 眼科研究

Objective The generate of posterior capsular opacification(PCO)is associated with the adhensive and migration of residual subcapsular lens epithelial cells(LECs).Titanium dioxide(TiO_2)nanometer is proved to have the ability of killing tumor cells and cultured bovine LECs.This study tried to observe the effects of TiO_2 nanometer thin film provoked by light on adhesiveness and migration of bovine LECs in vitro.MethodsThe fresh bovine lenses were obtained and cultured in DMEM containing 10% of newborn bovine serum.The second to fifth generation of cells were used in this experiment.The slide modified by TiO_2 photocatalyst film was prepared by sol-gel method.Cultured cells were seeded in filmed or unfilmed slides respectively and exposed to ultraviolet(wavelength 365 nm)for 20 or 40 minutes.The contact angle between water drop and slide was measured by droping method and the cells adhered to slides were calculated after 24 and 48 hours of culture.The growth status and migration distance of bovine LECs were assayed and compared between filmed and unfilmed groups.ResultsThe contact angle between water drop and slide was 0°±2°and 18.825°±2.342° in filmed and unfilmed group respectively,indicating a obviously smaller contact angle in TiO_2 filmed group than unfilmed one.The numbers of bovine LECs adhered to filmed slide was considerably reduced in TiO_2 filmed group compared with unfilmed group in different UVA exposure time(t_(0 min)=5.492,P=0.001;t_(20 min)=6.031,P=0.000;t_(40 min)=6.828,P=0.000).However,no significant difference was found in the numbers of adhensive cells among 3 UVA irradiation time points(F=1.278,P=0.297).The migration distance of the cells was significantly shorter in TiO_2 filmed group in comparison with unfilmed group in 24 and 48 hours after UVA irradiation(F_(group)=14.965,P=0.000;F_(time)=38.033,P=0.000).ConclusionThe TiO_2 nanometer thin film is characterized by the superhydrophilic property.So it can effectively impede the adhesion and migration of bovine LECs in vitro.

Differences in effects of Zuojin Pills and its similar formulas on Wei cold model in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the effects of Zuojin Pills and its similar formulas on the stomach cold syndrome in a Wei cold model in rats.</p><p><b>METHODS</b>The rat Wei cold model was established by intragastric administration of glacial NaOH, and the gastric mucosa injury indices, together with the levels of motilin and gastrin in the stomach, were determined. The preventive and curative effects of Zuojin Pills and its similar formulas on gastric mucosa injury were investigated.</p><p><b>RESULTS</b>Zuojin Pills and its similar formulas could protect the gastric mucosa in the gastric cold model in rats at different levels. Fanzuojin Pills had the best effect in inhibiting gastric mucosa injury.</p><p><b>CONCLUSION</b>The different pharmacological effects of Zuojin Pills and its similar formulas in the rat gastric cold model were partially correlated with the degrees in cold and heat properties of the formulas.</p>

Effect of Radix isatidis polysaccharides on immunological function and expression of immune related cytokines in mice / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of Radix Isatidis polysaccharides (RIP) on the immunological function and expression of immune related cytokines in mice.</p><p><b>METHODS</b>Sixty mice were randomly divided into six groups, the normal group, cyclophosphamide (Cy) model group, Levamisole (positive control) group, RIP low, medium and high dose groups (0.08 g/kg, 0.16 g/kg, 0.32 g/kg, respectively), ten in each group. By detecting the value of abdominal macrophage phagocytic index, serum hemolysin (HC50), proliferation of lymphocytes and expression of related cytokines, interleukin (IL-2) and interferon gamma (INF-gamma), the effect of RIP on immunological function and its mechanism were studied.</p><p><b>RESULTS</b>RIP could improve the level of hemolysin in immunological function depressed mice. The results showed that the value of macrophage phagocytic index in the high dose RIP group increased from 1.11+/-0.13 to 1.42+/-0.26. The level of IL-2 and INF-gamma could be decreased by Cy to 38.12+/-6.88 ng/L and 139.23+/-29.87 ng/L, respectively, while RIP in high dose could increase the secretion of IL-2 and INF-gamma to 53.54+/-14.43 ng/L and 189.91+/-32.63 ng/L, respectively.</p><p><b>CONCLUSION</b>RIP could enhance non-specific immunological function, humoral immunity and cellular immunity in mice.</p>

Value of Histological Staining in Differential Diagnosis of Vitiligo / 대한피부과학회지

BACKGROUND: Vitiligo is a depigmented disorder, causing serious cosmetic problems for patients. In diagnostic and therapeutic aspects, vitiligo should be differentiated from other hypopigmented disorders as the therapeutic approach and prognosis are different for each disease. OBJECTIVE: This study aimed to compare the usefulness of several markers for melanocytes or melanin in differential diagnosis of vitiligo. METHODS: Twenty-eight patients were studied, who were diagnosed clinically as suffering from one of the following diseases: vitiligo, nevus depigmentosus, pityriasis alba, postinflammatory hypopigmentation, and idiopathic guttate hypomelanosis. Skin samples (frozen or paraffin-fixed) were obtained from depigmented patches and normal neighboring skin (control). Histological staining was performed by using Fontana-Masson, S-100, MART-1, and DOPA. The staining level of lesional skin was compared with that of normal skin. RESULTS: When the staining level of vitiligo was compared with that of others, vitiligo was significantly lower in Fontana-Masson (13.3+/-17.2% vs 44.4+/-23.7%), S-100 (49.5+/-14.9% vs 74.7+/-24.2%), MART-1 (7.4+/-8.7% vs 68+/-33.9%), and DOPA (9.5+/-11.3% vs 58.2+/-29.5%) (p<0.0001). CONCLUSION: MART-1 and DOPA are valuable markers in differential diagnosis of vitiligo. However, Fontana-Masson, a marker of melanin, had some limits in detecting melanocytes, and S-100 showed non-specific staining other than melanocytes.

Expression of Peroxiredoxin I in the Epidermis of Vitiligo / 대한피부과학회지

BACKGROUND: Although the pathogenesis of vitiligo isn't fully understood, a recent study demonstrates that oxidative stress plays an important role to induce vitiligo. Peroxiredoxin (Prx) is a novel peroxidase family to remove hydrogen peroxide using thioredoxin system, which is consisted of thioredoxin, thioredoxin reductase, and NADPH. OBJECTIVE: This study aimed to investigate the change of expression of Prx I to elucidate the role of oxidative stress in the pathogenesis of vitiligo. METHODS: Sample specimens were obtained from the lesional skin of vitiligo patients, and non-depigmented skin was obtained from the perilesional area as control samples. The skin samples were immediately frozen using liquid nitrogen, and then section samples were prepared to perform immunohistochemical staining with antibodies for Prx I. Some of the skin biopsy samples were used for primary culture of keratinocytes. Protein extracts from the expanded keratinocytes were prepared for Western blot analysis of Prx I. RESULTS: In vitiligo, the ubiquitous expression of Prx I in all layers of epidermis, which was also observed in the normal perilesional skin, was reduced in the depigmented lesion of vitiligo patients. The reduction of Prx I was remarkable from the lesions which were exposed to sunlight. Consistently, Prx I expression from the lesional keratinocytes were noticeably reduced in comparison with that from perilesional keratinocytes. CONCLUSION: Our results showing that Prx I is impaired in the epidermis of depigmented lesions of vitiligo patients suggest that oxidative stress is an important factor to induce vitiligo.

Construction of recombinant adenovirus including microdystrophin and expression in the mesenchymal cells of mdx mice / 生物工程学报

Construction of recombinant adenovirus, which contain human microdystrophin, and then transfection into mesenchymal cells( MSCs) of mdx mice were done, and genetically-corrected isogenic MSCs were acquired; the MSCs transplantation into the mdx mice was then done to treat the Duchenne muscular dystrophy( DMD). Microdystrophin cDNA was obtained from recombinant plasmid pBSK-MICRO digested with restrictive endonuclease Not I ; the production was inserted directionally into pShuttle-CMV. The plasmid of pShuttle-CMV-MICRO was digested by Pme I , the fragment containing microdystrophin was reclaimed and transfected into E. coli BJ5183 with plasmid pAdeasy-1. After screening by selected media, the extracted plasmid of positive bacteria was transfected into HEK293 cells with liposome and was identified by observing the CPE of cells and by the PCR method. Finally, MSCs of mdx mice were infected with the culture media containing recombinant adenovirus, and the expression of microdystrophin was detected by RT-PCR and immunocytochemistry. Recombinant adenovirus including microdystrophin was constructed successfully and the titer of recombinant adenovirus was about 5.58 x 10(12) vp/mL. The recombinant adenovirus could infect MSC of mdx mice and microdystrophin could be expressed in the MSC of mdx mice. Recombinant adenovirus including microdystrophin was constructed successfully, and the microdystrophin was expressed in the MSC of mdx mice. This lays the foundation for the further study of microdystrophin as a target gene to correct the dystrophin-defected MSC for stem cell transplantation to cure DMD.

Sibling brother and sister both with Duchenne muscular dystrophy / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the clinical and lab features of sibling brother and sister both with Duchenne muscular dystrophy (DMD).</p><p><b>METHODS</b>We conducted comprehensive clinical and lab investigations including the test of serum enzymes, electromyography (EMG), electrocardiography, color Doppler echocardiography, HE staining of skeletal muscles, immunohistochemical study of dystrophin and utrophin, multiple ligation probe amplification (MLPA) on exon 1-79 of dystrophin gene, and short tandem repeat-poly- merase chain reaction of CA repeats located in dystrophin gene.</p><p><b>RESULTS</b>These two patients were confirmed to suffer from DMD. They were characterized by typical features of DMD including typical clinical manifestations, increased serum enzymes, EMG presenting myogenic impairment, HE staining presentation belonging to DMD, negative dystrophin in brother, and inconstantly positive on the sarcolemma of sister. Furthermore, no deletion or duplication was found in the 1-79 exons of dystrophin gene. The suffering brother and sister carried the same maternal X chromosome.</p><p><b>CONCLUSIONS</b>Carriers of DMD gene show typical clinical and laboratory manifestations of DMD. Comprehensive examinations should be performed for such carriers.</p>

Extrauterine growth retardation and correlated factors in premature neonates / 中华儿科杂志

<p><b>OBJECTIVE</b>Poor growth is a common problem in premature neonates. No sufficient attention has been paid to the nutrition deficit and extrauterine growth retardation in premature neonates in China. The present study aimed to assess the incidences of intrauterine growth retardation (IUGR) and extrauterine growth retardation (EUGR) in premature neonates in Shanghai area and their correlated factors.</p><p><b>METHODS</b>Data of the neonates discharged between January 1, 2003 and December 31, 2004 from 5 hospitals (Xinhua Hospital, Shanghai Children's Medical Center, Shanghai Children's Hospital, Pediatric Hospital Affiliated to Fudan University, and Shanghai International Peace Maternity and Child Health Hospital) were reviewed. The criteria of exclusion were cases who died or had a malformed appearance, the mother had endocrine or metabolic diseases. The criteria for enrollment were (1) gestational age < 37 weeks, (2) admitted less than 24 hours after birth and discharged from the same hospital, (3) duration of hospitalization was > or = 7 days. The growth values on discharge of each patient were compared to the expected values based on the intrauterine growth data and postmenstrual day on discharge. Growth retardation was defined as measured growth values (weight, head circumference) < or = 10th percentile of the values (growth expectation based on estimated postmenstrual age). In each specific group, the number of neonates with < or = 10(th) percentile for each growth parameter was counted and the percentages of patients who had values < or = 10(th) percentile on birth and discharge were calculated. The growth curves used for assessing birth weight and head circumference for different gestational age neonates were those published in 1986. All the data were analyzed using the SPSS statistical software package. The risk factors for extrauterine growth retardation on weight and head circumference were estimated with logistic regression model.</p><p><b>RESULTS</b>The subjects included 1196 premature neonates in the five hospitals (734 boys and 462 girls). The incidence of IUGR was 22.7% and 19.2% assessed by weight and head circumference, respectively. The incidence of EUGR was 49.7% and 23.1% assessed by weight and head circumference, respectively. Assessment of IUGR in accordance with the birth weight and head circumference in the five hospitals showed no significant correlation between IUGR and non-IUGR by birth weight (chi(2) = 4.944) and head circumference (chi(2) = 0.017) respectively. Whereas the assessment of EUGR in accordance with weight and head circumference showed a significant correlation between EUGR and non-EUGR by weight on discharge (chi(2) = 28.109), but no significant correlation was found between EUGR and non-EUGR by head circumference on discharge (chi(2) = 0.275). In specific birth weight groups, the lower the weight, the higher the incidence of IUGR and EUGR by the weight and head circumference. The incidence of EUGR in VLBWI was 78.9% and 50.0% assessed by weight and head circumference, respectively. Assessed by weight, significant correlation was observed between EUGR and non-EUGR in birth weight (t = 18.674), hospitalization duration (Z = -8.790) and the median number of day for total using EN (Z = -4.650); but by the head circumference, significant correlation was observed in head circumference at birth (t = 9.555), hospitalization duration (Z = -3.930) and the median number of day for total using EN (Z = -3.004). The relationship between EUGR and some risk factors was analyzed with Logistic regression model. Assessed by the weight, the following 4 factors were related to EUGR: sex (chi(2) = 10.351), gestation age at birth (chi(2) = 56.275), birth weight (chi(2) = 102.126) and different hospital (chi(2) = 4.773). Assessed by the head circumference, the following 2 factors were correlated: gestation age at birth (chi(2) = 10.322) and head circumference (chi(2) = 10.620).</p><p><b>CONCLUSIONS</b>This study showed that the incidence of EUGR in premature neonates was significantly higher than the data reported in other countries. The incidence of EUGR increased with the decreased birth weight. Different nutritional support had influence on EUGR.</p>

Transfection and in vitro expression of human microdystrophin gene in rat mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector of human microdystrophin gene and observe its expression in rat mesenchymal stem cells (rMSCs) in vitro.</p><p><b>METHODS</b>The plasmid PBSK-MICRO containing human microdystrophin cDNA was digested by restriction endonuclease, and the resultant microdystrophin fragment was inserted into the NotI site of pcDNA3.1(+) to prepare the eukaryotic expression vector-pcDNA3.1(+)/ microdystrophin, which was identified by endonuclease digestion and sequencing. The recombinant plasmid was transfected into rMSCs via lipofectamine, and after G418 selection, the expression of microdystrophin was detected by RT-PCR and indirect immunofluorescence assay.</p><p><b>RESULTS</b>Microdystrophin gene fragment was correctly inserted into the plasmid pcDNA3.1(+), as conformed by sequencing and digestion with Not I and Hind III. The total mRNA of the transfected rMSCs was extracted and microdystrophin mRNA expression was found in the cells by RT-PCR. Indirect immunofluorescence assay for the protein expression of microdystrophin showed bright red fluorescence in the transfected rMSCs.</p><p><b>CONCLUSION</b>Eukaryotic expression plasmid pcDNA3.1(+)/microdystrophin has been constructed successfully and microdystrophin can be expressed in transfected rMSCs in vitro, which may facilitate further research of Duchenne muscular dystrophy treatment by genetically modified allogeneic stem cell transplantation.</p>

Dystrophin expression and pathology of diaphragm muscles of mdx mice after xenogenic bone marrow stem cell transplantation / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of bone marrow stem cell transplantation (BMT) on the diaphragm muscles of mdx mice, a mouse model of Duchenne muscular dystrophy (DMD).</p><p><b>METHODS</b>The bone marrow-derived stem cells form male SD rats was transplanted through the tail vein into 18 female 8-week-old mdx mice, which were sacrificed at 4, 8 and 12 weeks after BMT (6 at each time point), respectively. The diaphragm muscles of the mice were subjected to HE staining, immunofluorescence detection of dystrophin, reverse transcription (RT)-PCR analysis of dystrophin mRNA transcripts and PCR analysis of Sry (sex-determining region on the Y chromosome) gene, with age-matched female C57 mice and untreated mdx mice as the controls.</p><p><b>RESULTS</b>The proportion of centrally nucleated fibers (CNF) in the diaphragm muscle of the recipient mdx mice was (15.58+/-0.91) %, (12.50+/-1.87) % and (10.17+/-1.17) % at 4, 8 and 12 weeks after BMT, respectively, significantly smaller than that of untreated mdx mice [(19.5+/-1.87) %], and the fibers after BMT showed less inflammatory infiltration. Compared with the untreated mice, the recipient mdx mice showed green fluorescence on significantly more diaphragm muscle cell membranes [with the proportion of dystrophin-positive fibers of (1.00+/-0.32) %, (6.00+/-1.05) % and (11.92+/-1.11) % at 4, 8, and 12 weeks after BMT]. RT-PCR of dystrophin mRNA also demonstrated significantly higher relative levels of dystrophin in the recipient mdx mice (0.19+/-0.05, 0.26+/-0.06 and 0.36+/-0.04 at 4, 8 and 12 weeks after BMT) than in untreated mdx mice, and Sry gene was present in the recipient mice.</p><p><b>CONCLUSION</b>BMT can partially restore dystrophin expression and ameliorate the pathology in the diaphragm muscles of mdx mice, and has great potential to produce general therapeutic effect in patients with DMD.</p>